Neurochemistry

Protective effect of a 3 kDa peptide obtained from beef myofibrillar protein using alkaline-AK on neuronal cells Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Seung Yun Lee, Sun Jin Hur Abstract The protective effect of two 3 kDa peptide fractions (AK3KF1 and AK3KF2), obtained from beef myofibrillar protein using an inexpensive enzyme (alkaline-AK) on human neuronal cells (SH-SY5Y) against H2O2-induced apoptosis was investigated. These peptides were isolated and further separated by fast protein liquid chromatography (FPLC), and their protective effect against H2O2-mediated cell death was measured by determining cell viability, nitric oxide (NO) production, mitochondrial membrane potential (MMP), apoptosis, morphological changes in cell nuclei, and in vitro antioxidant assays. The results indicated that treatment with peptide fractions increased cell viability and MMP, and decreased NO production, fragmentation of cell nuclei, and apoptosis in H2O2-treated SH-SY5Y cells. This is the first study to report neuroprotective effects of a peptide obtained from beef myofibrillar protein. The peptide sequence was identified as Thr-Gln-Lys-Lys-Val-Ile-Phe-Cys (TQKKVIFC). Thus, these findings suggest that TQKKVIFC can prevent neuronal cell death and could be useful in preventing neurodegenerative diseases. Graphical abstract

Repeated exposure to methiopropamine increases dendritic spine density in the rat nucleus accumbens core Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Wen Ting Cai, Hyung Shin Yoon, Sooyeun Lee, Jeong-Hoon Kim Abstract Repeated exposure to classical psychomotor stimulants, like amphetamine (AMPH), produces locomotor sensitization and accompanied structural plasticity of dendritic spines in the nucleus accumbens (NAcc). Following our previous report that repeated administration of methiopropamine (MPA), a structural analog to meth-AMPH, produces locomotor sensitization, it was examined in the present study whether this behavioral change also accompanies with structural plasticity in the NAcc in a similar way to AMPH. A week after adeno-associated viral vectors containing enhanced green fluorescent protein (eGFP) were microinjected into the NAcc core, rats were repeatedly injected with saline, AMPH (1 mg/kg, IP), or MPA (5 mg/kg, IP) once every 2–3 days for a total of 4 times. Two weeks after last injection, all rats were perfused and their brains were processed for immunohistochemical staining. The image stacks for dendrite segments of medium spiny neuronal cells in the NAcc core were obtained and dendritic spines were quantitatively analyzed. Interestingly, it was found that the number of total spine density, with thin spine as a major contributor, was significantly increased in MPA compared to saline pre-exposed group, in a similar way to AMPH. These results indicate that MPA, a novel psychoactive substance, has similar characteristics with AMPH in that they both produce structural as well as behavioral changes, further supporting MPA's dependence and abuse potential.

Ammonium induced dysfunction of 5-HT2B receptor in astrocytes Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Tingting Yue, Baoman Li, Li Gu, Jingyang Huang, Alexei Verkhratsky, Liang Peng Abstract Previously we reported that gene expression of astrocytic 5-HT2B receptors was decreased in brains of depressed animals exposed to chronic mild stress (CMS) (Li et al., 2012) and of Parkinson's disease (Song et al., 2018). Depression is also one of the psychiatric symptoms in hyperammonemia, and astrocyte is a primary target of ammonium in brain in vivo. In the present study, we have used preparations of the brains of urease-treated mice and ammonium-treated astrocytes in culture to study gene expression and function of 5-HT2B receptors. The urease-treated mice showed depressive behaviour. Both mRNA and protein of 5-HT2B receptors were increased in the brains of urease-treated mice and in ammonium-treated cultured astrocytes. Further study revealed that mRNA and protein expression of adenosine deaminase acting on RNA 2 (ADAR2), an enzyme catalyze RNA deamination of adenosine to inosine was increased in the brains of urease-treated mice and in ammonium-treated cultured astrocytes. This increase in ADAR2 induced RNA editing of 5-HT2B receptors. Cultured astrocytes treated with ammonium lost 5-HT induced Ca2+ signalling and ERK1/2 phosphorylation, indicating dysfunction of 5-HT2B receptors. This is in agreement with our previous observation that edited 5-HT2Breceptors no longer respond to 5-HT (Hertz et al., 2014). Ammonium effects are inhibited by ADAR2 siRNA in cultured astrocytes, suggesting that increased gene expression and editing and loss of function of 5-HT2Breceptors are results of increased activity of ADAR2. In summary, we have demonstrated that functional malfunction of astrocytic 5-HT2B receptors occurs in animal models of major depression, Parkinson depression and hepatic encephalopathy albeit via different mechanisms. Understanding the role of astrocytic 5-HT2B receptors in different pathological contexts may instigate development of novel therapeutic strategies for treating disease-specific depressive behaviour.

Tat-HSP70 protects neurons from oxidative damage in the NSC34 cells and ischemic damage in the ventral horn of rabbit spinal cord Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Woosuk Kim, Hyun Jung Kwon, Hyo Young Jung, Dae Young Yoo, Seung Myung Moon, Dae Won Kim, In Koo Hwang Abstract Heat shock protein 70 (HSP70) is an ATP-dependent molecular chaperone, and it has been shown that its levels increase after exposure to various types of stress, including ischemia. In the present study, we investigated the effects of HSP70 against H2O2-induced neuronal stress in NSC34 cells and against spinal cord ischemia in rabbits. Tat-HSP70 proteins facilitated the intracellular delivery of HSP70 into the NSC34 cells and enabled them to cross the blood-brain barrier in the rabbit spinal cord. Tat-HSP70 was effectively transduced into NSC34 cells in a concentration- and time-dependent manner, while control-HSP70 protein could not be delivered intracellularly at any concentration or time after treatment. Treatment with Tat-HSP70 reduced the generation of reactive oxygen species and cell death induced by H2O2, while the control-HSP70 did not show any significant effect on the NSC34 cells exposed to H2O2. In rabbit spinal cord, the administration of Tat-HSP70 showed significant amelioration of neurological defects and neuronal death in the ventral horn of spinal cord. In addition, Tat-HSP70 treatment significantly reduced lipid peroxidation and increased Cu, Zn-superoxide dismutase activities in the spinal cord, but glutathione peroxidase and Mn-superoxide dismutase activities remained unchanged. These results suggest that Tat-HSP70, not control-HSP70, decreases cell damage by reducing oxidative stress in NSC34 cells and rabbit spinal cord, and it can be employed for the reduction of neuronal damage caused after spinal cord ischemia.

Gut dysbiosis and lack of short chain fatty acids in a Chinese cohort of patients with multiple sclerosis Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Qin Zeng, Junli Gong, Xiyuan Liu, Chen Chen, Xiaobo Sun, Huijuan Li, Yifan Zhou, ChunPing Cui, Yuge Wang, Yu Yang, Aimin Wu, Yaqing Shu, Xueqiang Hu, Zhengqi Lu, Song Guo Zheng, Wei Qiu, Yongjun Lu Abstract Background Recent studies, mostly conducted in Western countries, showed that gut microbes are involved in the pathogenesis of multiple sclerosis (MS). Objective The aim of this study was to investigate whether gut dysbiosis is relevant to the initiation and progression of MS in a Chinese population. Methods Next-generation sequencing (NGS) and gas chromatography (GC) were integrated and used to compare the fecal bacterial communities and the short-chain fatty acid (SCFA) levels among relapsing-remitting MS (RRMS) patients (n = 34), neuromyelitis optica spectrum disorder (NMOSD) patients (n = 34), and healthy controls (HCs) (n = 34). T-cell profile analyses were performed by flow cytometry for MS patients and matched controls (n = 12). Results (1) The gut microbiome of MS patients was characterized by an increase of Streptococcus and a decrease of Prevotella_9; additionally, compared to NMOSD patients, Prevotella_9 was found to be much more abundant in MS patients. (2) A striking depletion of fecal acetate, propionate, and butyrate was observed in MS patients compared to HCs. (3) The abundance of Streptococcus was negatively correlated with the proportion of pTregs (P < 0.05) and positively correlated with Th17 cells (P < 0.05) in the peripheral blood, while the abundance of Prevotella_9 was negatively correlated with the Th17 cell frequency (P < 0.01), and the fecal SCFA level was positively correlated with pTreg frequency (P < 0.05). Conclusions Gut dysbiosis and a lack of SCFAs exist in Chinese MS patients, which might be related to an aberrant immune response of MS; this relationship may have a diagnostic and therapeutic value for patients with MS.

ML171, a specific inhibitor of NOX1 attenuates formalin induced nociceptive sensitization by inhibition of ROS mediated ERK1/2 signaling Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Sanjay Kumar, Ajeet Kumar Singh, Manjula Vinayak Abstract Reactive oxygen species (ROS) have a key role in different etiologies of pain. At sub-cellular level, mitochondria and plasma membranes have been identified as endogenous sources of ROS required for pain generation. NADPH oxidase (NOX) is the main contributor of membrane associated ROS generation. Out of 7 isozymes, NOX1, NOX2 and NOX4 are reported to be associated with nociceptive sensitization. Therefore, it has been hypothesized that specific inhibition of the NOX isozymes could be putative strategy for treatment of pain. However, unavailability of specific inhibitors was the biggest obstacle to test this hypothesis. Here, we investigated anti-nociceptive potential of a newly identified specific NOX1 inhibitor ML171 in formalin induced inflammatory pain. ML171 administration decreased the paw lickings and flinching response in both phases of formalin test. Behavioral response was supported with decreased activation of c-Fos in spinal dorsal horn. The increased level of total NOX activity, ROS and pERK1/2 in dorsal root ganglion (DRG) and spinal dorsal horn of formalin induced nociception were reversed by ML171 administration. ML171 also inhibited the upregulated Tumor necrosis factor receptor 1 (TNFR1) expression in DRG, whereas did not show any effect in spinal dorsal horn which was unaltered after formalin insult. The study for the first time depicts anti-nociceptive potential of ML171 via regulation of ROS mediated ERK1/2 signaling by inhibition of NOX1 activity.

Behavioral alterations induced by post-weaning isolation rearing of rats are accompanied by reduced VGF/BDNF/TrkB signaling in the hippocampus Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): M. Chmelova, L. Balagova, M. Marko, S. Vrankova, M. Cebova, D. Jezova, I. Riecansky, N. Hlavacova Abstract Post-weaning social isolation has been shown to be a relevant animal model for studying the mechanisms underlying psychopathological states induced by early-life stressful experiences. Besides extensively studied brain-derived neurotrophic factor (BDNF) and tyrosine receptor kinase B (TrkB) receptor, increasing attention is being given to a neuropeptide precursor VGF (non-acronymic). Several lines of evidence indicate an interplay between the neurotrophins and nitric oxide signaling. This study investigated the long-term consequences of post-weaning social isolation on behavior, VGF/BDNF/TrkB pathway and two isoforms of nitric oxide synthase (NOS) in the hippocampus and examined whether these effects were sex-specific. Male and female Sprague-Dawley rats were reared either in social isolation or social groups from postnatal day 21 for 9 weeks (n = 12–15/group and sex). Post-weaning social isolation induced impairments in sensorimotor gating and increased anxiety-like behavior in rats of both sexes. These behavioral alterations were accompanied by attenuated gene expression of VGF and TrkB receptor in the hippocampus. Isolation-induced reduction in VGF gene expression was more evident in male isolates. Similar changes were found in neuronal NOS (nNOS) gene expression with reduced mRNA levels in male isolates. Gene expression of BDNF and inducible NOS was not influenced by isolation rearing or sex. In addition, sex-specific patterns of VGF and nNOS gene expression in the hippocampus with higher mRNA levels in males than in females were revealed. The present study demonstrates a relationship between nNOS, VGF, BDNF, and TrkB confirming a link between nitric oxide and neurotrophins signaling pathways. Our findings indicate that long-term post-weaning social isolation alters signaling via VGF/BDNF/TrkB and nNOS that could interfere with neurodevelopmental processes which may contribute to pathological behavioral symptoms in adulthood. Future studies are needed to support this suggestion since the direct mechanistic link has not been approached in this study. Graphical abstract

Acute lysine overload provokes marked striatum injury involving oxidative stress signaling pathways in glutaryl-CoA dehydrogenase deficient mice Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Alexandre Umpierrez Amaral, Bianca Seminotti, Janaína Camacho da Silva, Francine Hehn de Oliveira, Rafael Teixeira Ribeiro, Guilhian Leipnitz, Diogo Onofre Souza, Moacir Wajner Abstract Glutaric acidemia type I (GA I) is a neurometabolic disorder of lysine (Lys) catabolism caused by glutaryl-CoA dehydrogenase (GCDH) deficiency. Patients are susceptible to develop acute striatum degeneration during catabolic stress situations whose underlying mechanisms are not fully established. Thus, in the present work we investigated the effects of a single intrastriatal Lys administration (1.5–4 μmol) to 30-day-old wild type (WT) and GCDH deficient (Gcdh−/−) mice on brain morphology, neuronal injury, astrocyte reactivity and myelin structure, as well as signaling pathways of redox homeostasis. We observed a marked vacuolation/edema in striatum and at higher doses also in cerebral cortex of Gcdh−/−, but not of WT mice. Lys also provoked a reduction of NeuN and synaptophysin, as well as an increase of astrocytic GFAP, in the striatum of Gcdh−/− mice, indicating neuronal loss and astrocyte reactivity. Furthermore, we verified an increase of Nrf2 and NF-κB expression in the nuclear fraction, and a decrease of heme oxygenase-1 (HO-1) content in the striatum of Lys-injected Gcdh−/− mice, implying disruption of redox homeostasis. Finally, it was found that Lys provoked alterations of myelin structure reflected by decreased myelin basic protein (MBP) in the cerebral cortex of Gcdh−/−mice. Taken together, the present data demonstrate neuronal loss, gliosis, altered redox homeostasis and demyelination caused by acute Lys overload in brain of Gcdh−/− mice, supporting the hypothesis that increased brain concentrations of glutaric and 3-hydroxyglutaric acids formed from Lys may be responsible for the acute brain degeneration observed in GA I patients during episodes of metabolic decompensation.

Neuroprotection mediated by remote preconditioning is associated with a decrease in systemic oxidative stress and changes in brain and blood glutamate concentration Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Jana Jachova, Miroslav Gottlieb, Miroslava Nemethova, Lubica Macakova, Martin Bona, Petra Bonova Abstract It has been shown that ischemia of remote organs can generate resistance to ischemic conditions within sensitive brain tissues. However, only limited information about its mechanism is available. In the present paper, we used hind-limb ischemia by tourniquet to generate early remote ischemic tolerance in rats. The main objective was to investigate the role of glutamate in the process of neuroprotection and discover parameters that are affected in the blood of ischemia-affected animals. Our results showed that pretreatment with a hind-limb tourniquet caused a decrease in neurodegeneration by about 30%. However, we did not observe neurological deficit recovery. When compared to ischemia, glutamate concentration decreased in all observed brain regions (cortex, CA1 and dentate gyrus of hippocampus), regardless of their sensitivity to blood restrictions. In contrast to this, the blood levels raised significantly from 26% to 29% during the first four days of postischemic reperfusion. Pretreatment of animals reduced systemic oxidative stress—as represented by lymphocytic DNA damage—by about 80%, while changes in blood antioxidant enzymes (catalase, superoxide dismutase) were not detected. With these data we can further hypothesize that hind-limb-tourniquet preconditioning could accelerate brain-to-blood efflux of glutamate which could positively impact neuronal survival of ischemia-affected brain regions. Moreover, remote preconditioning improved systemic oxidative stress and did not seem to be affected by enzymatic antioxidant defenses in the blood.

Dopamine receptor activation mitigates mitochondrial dysfunction and oxidative stress to enhance dopaminergic neurogenesis in 6-OHDA lesioned rats: A role of Wnt signalling Publication date: October 2019 Source: Neurochemistry International, Volume 129 Author(s): Akanksha Mishra, Sonu Singh, Virendra Tiwari, Swati Chaturvedi, M. Wahajuddin, Shubha Shukla Abstract Nigral dopaminergic (DAergic) cell degeneration and depletion of dopamine neurotransmitter in the midbrain are cardinal features of Parkinson's disease (PD). Dopamine system regulates different aspects of behavioural phenotypes such as motor control, reward, anxiety and depression via acting on dopamine receptors (D1-D5). Recent studies have shown the potential effects of dopamine on modulation of neurogenesis, a process of newborn neuron formation from neural stem cells (NSCs). Reduced proliferative capacity of NSCs and net neurogenesis has been reported in subventricular zone, olfactory bulb and hippocampus of patients with PD. However, the molecular and cellular mechanism of dopamine mediated modulation of DAergic neurogenesis is not defined. In this study, we attempted to investigate the molecular mechanism of dopamine receptors mediated control of DAergic neurogenesis and whether it affects mitochondrial biogenesis in 6-hydroxydopamine (6-OHDA) induced rat model of PD-like phenotypes. Unilateral administration of 6-OHDA into medial forebrain bundle potentially reduced tyrosine hydroxylase immunoreactivity, dopamine content in substantia nigra pars compacta (SNpc) and striatum region and impaired motor functions in adult rats. We found decreased D1 receptor expression, mitochondrial biogenesis, mitochondrial functions and DAergic differentiation associated with down-regulation of Wnt/β-catenin signalling in SNpc of 6-OHDA lesioned rats. Pharmacological stimulation of D1 receptor enhanced mitochondrial biogenesis, mitochondrial functions and DAergic neurogenesis that lead to improved motor functions in 6-OHDA lesioned rats. D1 agonist induced effects were attenuated following administration of D1 antagonist, whereas shRNA mediated knockdown of Axin-2, a negative regulator of Wnt signalling significantly abolished D1 antagonist induced impairment in mitochondrial biogenesis and DAergic neurogenesis in 6-OHDA lesioned rats. Our results suggest that dopamine receptor regulates DAergic neurogenesis and mitochondrial functions by activation of Wnt/β-catenin signaling in rat model of PD-like phenotypes.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Microscopy and Ultrastructure

Mitogen-activated protein kinase pathway: A critical regulator in tumor-associated macrophage polarization Thikryat Neamatallah Journal of Microscopy and Ultrastructure 2019 7(2):53-56 The notion that inflammation is a critical component of cancer has been researched extensively. Tumor-associated macrophages (TAMs) are among the inflammatory cells that greatly influence cancer. In the tumor microenvironment (TME), macrophages can either stimulate or inhibit tumorigenesis. TAMs that stimulate tumor cell proliferation (M2-phenotype) enrich the TME with growth factors and immunosuppressive molecules, whereas tumor inhibitory TAMs (M1-phenotype) initiate the immune response to dampen tumor progression. Shifting between phenotypes is controlled by several components of the TME. Targeting macrophages, specifically inhibiting M2 TAMs, has been introduced successfully in cancer immunotherapy. However, signaling mechanisms underlining TAM polarization are largely unknown. This review analyzed studies of the role of mitogen-activated protein kinase (MAPK) as a determinant of macrophage polarization. It is proposed that activation of MAPK, particularly extracellular signal-regulated kinase 1/2 and p38, might favor the differentiation into M2 TAMs. Thus, pharmacological modification of MAPK pathways will potentially offer exciting new targets in cancer immunotherapy.

Ultrastructure morphological characterization of different passages of rat dental follicle stem cells at In vitro culture Fakhri A Al-Bagdadi, Humberto M Barona, Eduardo Martinez-Ceballos, Shaomian Yao Journal of Microscopy and Ultrastructure 2019 7(2):57-64 Introduction: Stem cells play important roles in tissue renewal and repair. Tissue-derived stem cells have been demonstrated for their applications in tissue engineering and regenerative medicine. Expansion of primary stem cells isolated from tissues to a large quantity through in vitro culture is needed for application of the stem cells. However, it is known that tissue stem cells commonly reduce or lose their stemness properties during in vitro culture. In this study, we assessed ultrastructural changes of rat dental follicle stem cells (DFSCs) during in vitro culture. It is our attempt to explain the loss of stemness properties in cultured tissue-stem cells at the ultrastructural level. Method: DFSCs was isolated from first molars of Sprague Dawley rat pups and cultured in medium consisting of alpha-MEM plus 20% FBS. Cells were passaged at 1 to 3 ratio at 90% confluence, and collected at passages 3, 6, 7 and 9 for assessment of ultrastructure morphology by transmission electron microscopy. Results: Of the four passages (3, 6, 7, and 9) examined, dilated rough endoplasmic reticulum (RER) was abundant in Passage 3 but less so in Passages 6, 7, and 9. The dilated RER contained lipid in Passages 3, 7, and 9. The mono- and polyribosomes in Passages 3 and 6 were located between the mitochondria and the RER. Mono- and polyribosomes were abundant in Passage 7, although mainly monoribosomes were present in Passage 9. Membrane-bound glycogen granules were in vacuoles bulging off the cells in Passage 3. Some glycogen granules were grouped in the periphery of a stem cell in Passage 9. Nuclei shapes were irregular and mainly euchromatic in Passages 6, 7, and 9. The mitochondria were dark and scarce in Passage 9; irregular, small, and dark in Passage 7; and small and rounded in Passage 6, and they were spread in the cytoplasm away from the nucleus in Passage 3. Cell contacts were seen in Passages 6, 7, and 9. The ultrastructure morphology of the examined DFScs was not very different from the morphology criteria of the undifferentiated cells. Large vacuoles in Passage 3 were mainly at the periphery of the cell, with the small vacuoles in the cell center. Small vacuoles were scattered in the cell center of Passage 6 and the larger ones were observed at the cell's periphery. Conclusions: We observed the following ultrastructural changes: decreases of fine cell cytoplasmic processes, dilated cytoplasmic vacuoles, cytoplasmic pinocytotic vesicles, and nuclear heterochromatin with increasing cell passage number. Conversely, mean ratios of lipid globules, nuclear euchromatin, irregular nuclear shape, and cell contact between cells were increased with passage number. The observations may suggest an increase in committed cells among the population after long-term culture of DFSCs.

The biological and hematological effects of Echinacea purpurea L. Roots extract in the immunocompromised rats with cyclosporine Hala A H. Khattab, Seham K Abounasef, Haneen L Bakheet Journal of Microscopy and Ultrastructure 2019 7(2):65-71 Background: The immune system is the body's defense against foreign organisms and harmful chemicals. Cyclosporine A (CsA) is an immunosuppressant drug widely used. Echinacea purpurea root (EPR) extract is used as an immunostimulant plant. Aim of the Work: The present study aimed at evaluation of the EPR effects against the CsA immunosuppressive rat model. Material and Methods: Thirty-two male Wistar albino rats were randomly divided into control, CsA (immunosuppressive models), CsA + EPR (100 mg/kg/day orally), and CsA + EPR (200 mg/kg/day orally). The biological parameters regarding the food consumption were assessed including feed intake (FI), feed efficiency ratio (FER), and body weights (BW). In addition, the splenic specimens were assessed histopathology. The blood was collected for measuring the blood parameters. All the measured parameters were collected and statistically analyzed. The biological results indicated a significant decrease in BW, FI, and FER in rats treated orally with low and high EPR doses as compared to the control group. Results: The results displayed that the CsA induced a significant decrease in red blood cells (RBCs) and white blood cells (WBCs) count. Histopathologically, CsA induced a marked decrease in the cellularity of the white pulp with congested blood sinusoids of the red pulp together with significant depletion of periarteriolar lymphoid sheath. Both the high and low doses of EPR significantly reversed the altered RBCs and WBCs counts. Histopathologically, both the low and high doses of EPR displayed apparently increase in the periarteriolar area together with the persistence of the congestion of the red pulp blood sinusoids compared to CsA group, indicating partial amelioration of the structural changes. Conclusion: In a nutshell, the current findings revealed that EPR extract ameliorated the hematological changes. However, there was a partial correction of the CsA-induced microscopic changes of the rat spleen.

The awareness of pregnant patient about effect of antibiotics in pregnancy Rehab Alsaleh, Shahad Gari, Maram Gari Journal of Microscopy and Ultrastructure 2019 7(2):72-77 Background: Medication during pregnancy should be prescribed under caution as some medication has adverse effects on fetus health and they may be teratogenic. Antibiotics are widely used in pregnancy as a result of infections, adding that many pregnant women may administrate antibiotic without doctors' prescription. It is very important to assess the awareness of females about the effect of antibiotics during the pregnancy period. Aim: The aim of the study is to assess the awareness of pregnant patient about effect of antibiotics in pregnancy. Subjects and Methods: This study is cross-sectional study which was conducted in 2017 from September to October. Data were collected using interviewing questionnaire which investigated several variables including sociodemographics, clinical examination, clinical history, and clinical measurements. Results: Education and economic levels significantly affected the knowledge about reason for using antibiotic (P = 0.006, 0.002), using of antibiotic against what (P = 0.005, 0.000), education level affected the knowledge about the using of antibiotic without doctors' prescription (P = 0.01), and while economic level influenced knowledge about the effect of antibiotic (P = 0.01). Conclusion: There was good level of knowledge about using of antibiotic during pregnancy among females which was affected by economic and education levels.

Knowledge of and attitudes toward the use of anabolic-androgenic steroids among the population of Jeddah, Saudi Arabia Maha H Ahmed, Najla S Al-Saud, Abdulkader M Omar, Rania M Magadmi, Sabah M Hassan, Fatma M Al-Qudsi Journal of Microscopy and Ultrastructure 2019 7(2):78-83 Purpose: The aim of this study was to investigate the correlation between the use of anabolic-androgenic steroids (AASs) among the population of Jeddah, Saudi Arabia, and their knowledge and attitudes. Methods: This was a community-based, cross-sectional observational study. This study was conducted using a questionnaire that was distributed among the population during the period from February 3, 2018, to February 25, 2018. This questionnaire comprised 31 questions, designed to evaluate the knowledge and attitudes toward using AASs. Results: A total of 300 participants were enrolled in the study. The mean age of the population was 30.66 ± 9.2 years. Fourteen participants admitted using AASs, with a percentage of 4.7%, among whom 85.7% were male (P = 0.0005). Seventy-eight percent of AAS users believed that AASs do not cause tolerance when taken for a longtime (P = 0.023). However, the majority of both AAS users and nonusers did not agree on taking AASs for a longtime. Our results showed a strong correlation between not taking AASs and not consuming energy drinks (P = 0.0023). Half of our respondents exhibited poor knowledge regarding the side effects of AASs. The level of knowledge did not correlate with the use of AAS, gender, exercising, or consuming energy drinks. Conclusion: The results showed poor knowledge regarding using AASs among the population of Jeddah. Thus, we recommend having a national awareness program in order to prevent the possible side effects of misusing AASs.

Possible protective role of panax ginseng on cisplatin-induced hepatotoxicity in adult male albino rats (Biochemical and Histological Study) Abdulaziz Abdulrahman Alrashed, Eman Ali El-Kordy Journal of Microscopy and Ultrastructure 2019 7(2):84-90 Background: Cisplatin is one of the most effective chemotherapy antineoplastic drugs. Panax ginseng is a well-known medicinal herb and has a long history of medicinal use as a tonic to promote health. Aim: This work aimed to study the effect of ginseng on the liver damage induced by cisplatin in rats. It included biochemical and histological investigations. Materials and Methods: Twenty adult rats were divided into four equal groups. Group I served as control. Group II received ginseng orally (100 mg/kg/day) for 4 weeks. Group III animals were injected intraperitoneally with cisplatin in three equal doses (each 3.3 mg/kg) daily for 3 consecutive days. Group IV animals received ginseng together with cisplatin by the same previously mentioned methods and doses. Rats were sacrificed after 4 weeks, and blood samples and liver tissues were collected for biochemical and histological examinations. Results: Cisplatin-induced liver damage manifested biochemically by an increase in serum alanine aminotransferase and aspartate aminotransferase. Histologically, hepatocytes appeared with vacuolated cytoplasm and small dark-stained nuclei with dilatation of blood sinusoids as well as marked accumulation of collagen fibers around enlarged portal tracts. Administration of ginseng together with cisplatin improved the hepatic dysfunctions and damage caused by cisplatin. Conclusion: Ginseng has a protective role in the amelioration of cisplatin-induced hepatotoxicity.

Efficacy of Ginger (Zingiber officinale) in ameliorating streptozotocin-induced diabetic liver injury in rats: Histological and biochemical studies Mona R Alshathly Journal of Microscopy and Ultrastructure 2019 7(2):91-101 Ginger (Zingiber officinale) was reported to have an antioxidant, antidiabetic effect. This study was done to investigate its therapeutic effect against functional and structural alteration in liver of diabetic rat (intraperitoneal streptozotocin (STZ) in a dose of 60 mg/kg/bw). Thirty adult male rats (three-months-old and 250 g weight) were sorted into five groups (N=6). G1 used as control, G2 was diabetic rats without any treatment, G3 was diabetic rats given oral ginger in a dose of 500 mg/kg/bw, G4 was diabetic rats treated with metformin (500 mg/kg/bw) while G5 received ginger orally. The experiment lasts for six weeks, animals were anesthetized by ether, body weight was recorded for all animals. Blood was collected for further analysis of lipid profile, liver enzymes and total antioxidant. Liver was dissected, weighted and samples were processed for histopathological study. The results showed significant decrease of glaucous level and liver enzymes in ginger treated rats. Total antioxidant was preserved. Ginger lowered blood glucose, level, regained body weight and liver index to near normal values. Diabetes induced degenerative changes and micro-vesicular lipid deposition in hepatocytes with moderate portal area fibrosis. Ultrastructure study confirmed such changes beside demonstrating increased lipid deposition in fat storing cells. Ginger was found to ameliorate those changes in treated animals. Results were matching metformin effects. In conclusion, Ginger as a natural safe Herbal medication can be used to support liver functions in diabetic status.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Ethnopharmacology

Antidiabetic and hypolipidemic efficacy of skin and seed extracts of Momordica cymbalaria on alloxan induced diabetic model in rats Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Abbirami Elangovan, Abinaya Subramanian, Siva Durairaj, Jeyadevi Ramachandran, Dinesh Kumar Lakshmanan, Guna Ravichandran, Gayathri Nambirajan, Sivasudha Thilagar Abstract Ethnopharmacological relevance Momordica cymbalaria, a wild vegetable belongs to the Cucurbitaceae family, has long been used as a food and a remedy for diabetes mellitus in the Asian native medicinal system. Aim of the study This study aims to evaluate the efficacy of ethanolic extract of skin (EESK) and methanolic extract of seed (MESE) of M. cymbalaria (MC), for their hypoglycemic and hypolipidemic effects in alloxan induced diabetic rats. Materials and methods The diabetes induced rats were given skin and seed extracts at doses 250 and 500 mg/kg b.w. p.o. for 28 days. Alloxan monohydrate (120 mg/kg) was used to induce diabetes mellitus. Daily food and water intake were assessed. Blood glucose levels and body weights were measured every 7 days throughout the experiment. Antioxidant assays, different biochemical and glycemic parameters were evaluated. Histopathological studies on pancreas, liver and kidney were also studied. Results Treatment of EESK and MESE showed dose significant decrease in fasting blood glucose level (FBG) in experimental diabetic animals with significant reduction in food and water intake and increase in body weight. Findings confirmed the hypoglycemic and hypolipidemic effects of EESK and MESE in the experimental groups. The impaired glucose tolerance and altered activities of the hepatic enzymes such as AST, ALT and ALP levels of diabetic rats were significantly improved by the administration of EESK and MESE. Oral treatment with MC extract for 28 days demonstrated significant protective effects on the lipid profile, biochemical parameters and antioxidant levels. Besides, biochemical findings were supported by histopathological investigations. Conclusion These results suggest that the treatment with EESK and MESE of MC at a dose of 500 mg/kg b.w. have better protective effects against hyperglycemia, hyperlipidemia and oxidative stress generated during diabetes justifying the use of the plant in traditional systems of medicine. Graphical abstract

Neoline is the active ingredient of processed aconite root against murine peripheral neuropathic pain model, and its pharmacokinetics in rats Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Yohei Tanimura, Masato Yoshida, Kan'ichiro Ishiuchi, Masahiro Ohsawa, Toshiaki Makino Abstract Ethnopharmacological relevance Processed aconite root (PA), the root of Aconitum carmichaeli (Ranunculaceae), is a crude drug used in traditional Chinese or Japanese kampo medicine to treat pain associated with coldness. In our previous study, PA and its active ingredient, neoline, alleviated oxaliplatin-induced peripheral neuropathy in mice. Aim of the study The present study investigated the effects of PA on a murine peripheral neuropathy model induced by intraperitoneal injection of paclitaxel and partial ligation of the sciatic nerve (Seltzer model), and identified its active ingredients. Materials and methods PA powder (1 g/kg/day) was orally administered, and either neoline or benzoylmesaconine (10 mg/kg/day) was subcutaneously injected into the murine model. Mechanical hyperalgesia was evaluated via the von Frey filament method. PA extract was orally administered to rats; blood samples were chronologically collected, and the plasma concentrations of Aconitum alkaloids were measured. The contents of Aconitum alkaloids in commercial PA products were also measured. Results PA extract and neoline significantly attenuated the mechanical hyperalgesia induced by either paclitaxel or partial ligation of the sciatic nerve in mice. In the plasma samples of rats treated with PA extract, higher concentrations of benzoylmesaconine and neoline were apparent among Aconitum alkaloids. The contents of benzoylmesaconine and neoline varied among PA products with different processing procedures. Subcutaneous injection of benzoylmesaconine did not attenuate the hyperalgesia induced by each paclitaxel, partial ligation of the sciatic nerve, or oxaliplatin in mice. Conclusions The present results indicate that PA and its active ingredient, neoline, are promising agents for the alleviation of neuropathic pain. Neoline can be used as a marker compound to determine the quality of the PA products for the treatment of neuropathic pain. Graphical abstract

Ethnobotanical survey of medicinal plants in central Abyan governorate, Yemen Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Mohamed Al-Fatimi Abstract Ethnopharmacological relevance Traditional medicinal plant knowledge in Yemen is still only passed on orally from one generation to another. A few studies, mostly very limited in scope, have been dedicated to this indigenous knowledge so far. This paper presents the results of the first extensive field study in southern mainland Yemen, undertaken in three communities in the central region of Abyan governorate. The study is aimed to preserve the heritage of this indigenous knowledge and to explore and select local medicinal plants that promise high pharmacological efficacy for further pharmacological and phytochemical investigations. Methods 356 indigenous informants (273 males, 83 females) were interviewed about the modes of application and uses of the medicinal plants in the region. The ethnobotanical data were recorded by semi-structured face-to-face interviews, substantiated by specimen collections and taxonomic identifications and quantitative data analysis including informant consensus factor (ICF) and number of use-reports. Results In total, 195 medicinal plant species (170 wild and 25 cultivated) belonging to 138 genera and 55 flowering plant families, were recorded for the treatment of 155 different ailments classified into 16 categories. Apocynaceae (25 species), Fabaceae (18 species), Euphorbiaceae (16 species) and Asteraceae (14 species) were the most frequently used plant families. Paste (86) followed by unprepared plant exudates (77) and decoction (55) were the most common herbal preparation modes. Dermal application (51.8%) was the most common administration route. The highest number of use reports and the maximum number of used medicinal plants were recorded for treatment of skin and gastrointestinal ailments. Forty-five species have never been reported in the ethnomedicinal literature before. Plant species with the most citations were Aloe vacillans Forssk. (malaria), Solanum incanum L. (tooth decay), Caralluma awdeliana (Deflers) A. Berger (diabetes), Tribulus terrestris L. (kidney stones), Aristolochia bracteolata Lam. (snake poison), Hydnora abyssinica A.Br. (stomach ulcer), Indigofera oblongifolia Forssk. (urine retention) and Chrozophora oblongifolia (Delile) A. Juss ex Spreng (haemorrhoids). Conclusions Significant traditional knowledge of the uses of local medicinal plant species was recorded for the first time for central Abyan in particular and southern Yemen in general. The study shows that the medicinal plants still play an important role in the primary health care in the study area. The ethnobotanical results provide a basis for further pharmacological, biological, pharmacognostical, and phytochemical investigations. Graphical abstract

A lipid-soluble extract of Pinellia pedatisecta Schott enhances antitumor T cell responses by restoring tumor-associated dendritic cell activation and maturation Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Yumeng Wang, Haixia Huang, Sheng Yao, Guiling Li, Congjian Xu, Yang Ye, Suiqi Gui Abstract Ethnopharmacological relevance Pinellia pedatisecta Schott (PPS)is a traditional Chinese medicine functioning as reducing swelling and drying dampness. Pinellia pedatisecta Schott extract (PE) has been confirmed to suppress cervical tumor growth and modulate the antitumor CD4+T helper immunity towards Th1. Aims To explore the roles of PE in modulating tumor-associated dendritic cell (TADC) activation and function. Methods For in vivo studies, HPV+TC-1 mouse tumor models were conducted and treated with PE for 3 weeks (10 mg/kg/d or 20 mg/kg/day). The immune profiles of spleen, tumor-draining lymph nodes (TDLNs), tumor and serum were analyzed by flow cytometry and multiplexed bead-based immunoassay. For in vitro studies, TADCs were generated by tumor-conditioned medium and treated with PE solution. The maturation and function of TADCs were evaluated by flow cytometry, ELISA, mixed lymphocyte reaction (MLR) and cytotoxic T lymphocyte (CTL) assay. Furthermore, the effect of PE on SOCS1 pathway was examined by western blotting and real time PCR. Results PE upregulated the expression of major histocompatibility complex class II (MHCII) and costimulatory molecules CD80 and CD86 on TADCs and promoted IL-12 secretion from TADCs. In addition, PE-treated TADCs promoted the proliferation of CD4+ and CD8+ T cells and induced the differentiation of IFN-γ+CD4+ and GZMB+CD8+ T cells. PE-treated TADCs also elicited a more powerful antigen-specific cytotoxic T lymphocyte (CTL) response. Furthermore, PE treatment in vivo enhanced the proliferation, activated the functional ability (increased Ki67, CD137, GZMB or IFN-γ, TNF-α expression) and reversed the exhaustion (impaired CD95 or PD-1 expression) of antitumor T cells. Mechanistically, PE inhibited SOCS1-restrained JAK2 activation in TADCs. Conclusions PE efficiently restored the immature status of TADCs and enhanced their function as antigen-presenting cells to further elicit antitumor Th1 and CTL responses, suggesting that PE may be a potential immunomodulatory drug for cancer treatment. Graphical abstract

Binding of the polysaccharide from Acanthopanax giraldii Harms to toll-like receptor 4 activates macrophages Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Qingqing Li, Zhiting Chen, Zhilu Xu, Shaoyun Han, Huihui Hao, Jiang Wu, Fengxiang Sun, Xiaoyan Fu, Ruyue Li, Birong Zheng, Xiaoxiao Guo, Tongtong Zhang, Yong Chen Abstract Ethnopharmacological relevance The traditional Chinese medicine, Acanthopanax giraldii Harms, is commonly used to treat arthralgia due to wind, cold and dampness, as well as weakness in the feet and knees. Its other reported effects include eliminating flatulence, strengthening muscles and bones, and delaying aging. The polysaccharides in A. giraldii Harms are the major bioactive substances that confer the herb's antioxidant properties as well as anticancer and antiviral effects. Aims of the study To elucidate the underlying mechanism and signaling cascade involved in the homogeneous A. giraldii Harms polysaccharide II (AHP–II)–mediated immunomodulation of mice macrophages. Materials and methods The phagocytosis of neutral red and the production of nitric oxide, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), were measured to determine AHP–II–induced macrophage activation. Confocal microscopy and flow cytometry were used to confirm the binding of AHP-II to macrophages. The involvement of Toll-like receptor (TLR) 4 in AHP–II–induced macrophage activation was demonstrated using antibody blocking and macrophages from C3H/HeJ TLR4-mutant mice. Western blotting was used to map AHP–II–induced downstream signaling pathways. Results AHP-II increased the phagocytosis of macrophages and the release of nitric oxide, IL-6 and TNF-α cytokines. Direct, saturable and reversible binding of AHP-II to macrophages was observed, while it can be inhibited by the anti-TLR4 antibody. In addition, the presence of the anti-TLR4 antibody inhibited AHP–II–induced macrophage IL-6 and TNF-α production in the peritoneal macrophages of C3H/HeJ mice. Moreover, AHP–II–TLR4-stimulated macrophages activate the downstream intracellular ERK and JNK/nuclear factor (NF)-κB signaling pathways. In addition, the AHP–II–mediated regulation of IL-6 and TNF-α production from macrophages was greatly affected by specific ERK, JNK and NF-κB inhibitors. Conclusion Our study elucidated the immunomodulatory mechanism of AHP-II in macrophage activation and identified TLR4 as the main receptor coordinating AHP-II binding. Our findings suggest AHP-II may be used as a novel immunopotentiator for medical purposes. Graphical abstract

Utilization trends in traditional Chinese medicine for acute myocardial infarction Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Chung-Yen Lu, Pei-Chin Lu, Pei-Chun Chen Abstract Ethnopharmacological relevance Among heart diseases, acute myocardial infarction (AMI) is the most serious and life-threatening emergency. In Taiwan, heart disease has consistently ranked second among the top 10 leading causes of death since 2007, second only to malignant tumors; however, population-based studies on the use of traditional Chinese medicine (TCM) in AMI cases are limited. Aim of the study: This study investigated the characteristics of TCM users and prescriptions of TCM, and their differences between two cohorts of patients with AMI, identified 10 years apart. Materials and methods A cross-sectional study was conducted using the Taiwan National Health Insurance claims database. From among two random sample of 1 million beneficiaries selected from the claims database, we identified two cohorts of patients with first hospitalization for AMI in between 2000–2001 and 2010–2011. Patients who had received TCM therapy within one year after hospital discharge were defined as TCM users, whereas, all the other patients with AMI were considered non-users of TCM. We compared the characteristics of TCM use and the patterns of prescriptions between the two cohorts. Results The proportion of patients receiving TCM care was similar between the two AMI cohorts; 20% (85/418) of the patients were diagnosed in 2000–2001 and 21% (169/817) in 2010–2011. In the 2010–2011 AMI cohort, the proportion of men was smaller among TCM users than non-users, and TCM users were less likely to have hyperlipidemia. Among TCM users, the most frequently prescribed herb was Dan-shen (Salvia miltiorrhiza Bunge, Salvia root) in both cohorts. The most commonly used Chinese herbal formulations were Xue-Fu-Zhu-Yu-Tang (Blood Mansion Dispel Stasis) for the 2000–2001 cohort and Zhi-Gan-Cao-Tang (Honey-Fried Licorice Decoction) for the 2010–2011 cohort. Conclusions This study revealed the differences in the prescription frequency of Chinese herbal formulation among the two cohort of patients with AMI, suggesting that the practice of prescribing TCM has evolved from post-antique formula to classical remedies during the 10 years evaluated. Further investigations are needed to evaluate if the change in the utilization of Chinese herbal formulations impacts the effectiveness of the treatment. Graphical abstract

Anxiolytic- and anxiogenic-like effects of Montanoa tomentosa (Asteraceae): Dependence on the endocrine condition Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Erika Estrada-Camarena, Isabel Sollozo-Dupont, Dannia Islas-Preciado, María Eva González-Trujano, Miguel Carro-Juárez, Carolina López-Rubalcava Abstract Ethnopharmacological relevance Montanoa tomentosa Cerv. (MT) is a native plant from Mexico used in traditional medicine as a remedy for reproductive impairments and relaxing effects. In previous studies, it has been shown that the endocrine state could modify the antianxiety-like actions of anxiolytic compounds. Although women are the primary user of MT, no studies have evaluated the potential impact of the endocrine milieu on its anti-anxiety actions. Aims of the study:Ascertain the antianxiety effects of M. tomentosa in rats with different hormonal conditions, and to analyze the participation of the GABAA receptor in ovariectomized rats treated with MT. Materials and methods The animal model of anxiety used was the elevated plus-maze (EPM). Rats' endocrine conditions were: a) Low hormone levels (rats in diestrus I and II phases); b) High hormone levels (proestrus/estrus phases); c) No hormones (ovariectomized rats); and d) Rats under progesterone withdrawal (PW). To evaluate the participation of the GABAA receptor in the anxiolytic-like action of MT the antagonist picrotoxin was used. Results Results showed that MT induced dose-dependent anxiolytic-like actions in rats with low hormone level conditions. Also, MT reduced anxiety-like behavior in female rats under PW, in contrast to diazepam which was ineffective. MT's anxiolytic-like effect was blocked by picrotoxin, suggesting the participation of the GABAA receptor complex. However, increased anxiety-like behavior was observed in rats with a high hormone level condition and low doses of MT. Conclusions Beneficial anxiolytic-like actions of MT are observed under low hormone conditions, particularly in the PW challenge (a condition that can be related to a premenstrual period). Furthermore, the participation of the GABAAreceptor is evidenced. However, hormonal variations could induce the opposite effects, hence women should be cautious. Graphical abstract

Hazard assessment of Maerua subcordata (Gilg) DeWolf. for selected endpoints using a battery of in vitro tests Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Mebrahtom Gebrelibanos Hiben, Lenny Kamelia, Laura de Haan, Bert Spenkelink, Sebastiaan Wesseling, Jacques Vervoort, Ivonne M.C.M. Rietjens Abstract Ethnopharmacological relevance Maerua subcordata (Gilg) DeWolf is a medicinal and wild food plant growing mainly in east Africa. Especially its root tuber is widely used in traditional medicine to treat several infectious and chronic diseases but also in some toxicity implications like use as abortifacient. Aim of the study the present study applied in silico and in vitro tests to identify possible hazards of M. subcordata (fruit, leaf, root, seed) methanol extracts focussing on developmental toxicity. Materials and methods Ames test, estrogen receptor alpha (ERα) assay, aryl hydrocarbon receptor (AhR) assay, embryonic stem cell test (EST), and zebrafish embryotoxicity test (ZET) were employed. Besides, a Derek Nexus toxicity prediction was performed on candidate structures obtained from metabolomics profiling of the extracts using liquid chromatography coupled to multistage mass spectroscopy (LC/MSn) and a MAGMa software based structural annotation. Results Glucosinolates, which degrade to isothiocyanates, and biogenic amines were among the candidate molecules identified in the extracts by LC/MSn - MAGMa software structural annotation. Isothiocyanates and some other candidate molecules suggested a positive mutagenicity alert in Derek toxicity predictions. All the extracts showed negative mutagenicity in the Ames test. However, the Derek predictions also identified endocrine and developmental toxicity as possible endpoints of concern. This was further assessed using in vitro tests. Results obtained reveal that leaf extract shows AhR and ERα agonist activities, inhibited differentiation of ES-D3 stem cells into contracting cardiomyocytes in the EST (p < 0.001) as well as inhibited hatching (p < 0.01) and showed acute toxicity (p < 0.01) in the ZET. Also, the fruit extract showed toxicity (p < 0.05) towards zebrafish embryos and both fruit and seed extracts showed AhR agonist activities while root extract was devoid of activity in all in vitro assays. Conclusion The leaf extract tests positive in in vitro tests that may point towards a developmental toxicity hazard. The current evaluations did not raise concerns of genotoxicity or developmental toxicity for the fruit, seed and root extracts. This is important given the use of especially these parts of M. subcordata, in traditional medicine and/or as (famine) food. Graphical abstract

The effect of isoflavaspidic acid PB extracted from Dryopteris fragrans (L.) Schott on planktonic and biofilm growth of dermatophytes and the possible mechanism of antibiofilm Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Haoqi Lin, Xueping Liu, Zhibin Shen, Wanqiu Cheng, Zhijun Zeng, Yanfen Chen, Chunping Tang, Tao Jiang Abstract Ethnopharmacological relevance Dryopteris fragrans (L.) Schott (D. fragrans), a deciduous perennial herb, has been traditionally used for treatment of various skin diseases in Heilongjiang province of China for many years. Phloroglucinol derivatives extracted from D. fragrans were the most effective fraction against dermatophytes. Isoflavaspidic acid PB is a typically phloroglucinol derivative which extracted from D. fragrans and has been reported to exert anti-fungal activities against several dermatophytes. Aim of the study. This study aimed to evaluate anti-fungal and anti-biofilm activity of isoflavaspidic acid PB on planktonic and biofilm growth of dermatophytes and explore possible mechanisms of anti-biofilm. Materials and methods Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of isoflavaspidic acid PB against 25 isolates of dermatophytes were determined by the Clinical and Laboratory Standards Institute (CLSI) M38-A2 method. The effects of isoflavaspidic acid PB on dermatophytes biofilm formation and pre-formed biofilm were assessed by 2.3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[carbonyl (phenylamino)]-2H-tetrazolium hydroxide (XTT) assay. Morphology of mature biofilm were observed by Scanning Electron Microscope (SEM). Biomass, exopolysaccharide and ergosterol content of mature biofilm were analyzed by gravimetric analysis, anthranone sulfuric acid method and Ultra Performance Liquid Chromatography (UPLC) assay respectively. Result The MIC and MFC ranges of isoflavaspidic acid PB against 25 isolates of dermatophytes were 20–80 μg/mL and 40–80 μg/mL respectively. Isoflavaspidic acid PB (2 MIC) inhibited not only Trichophyton biofilm formation (54.8% ∼ 81.2%) but also the metabolic activity of mature biofilm (20.7% ∼ 44.2%). The result of SEM showed that isoflavaspidic acid PB (8 MIC) could destroy the morphology of hyphae seriously. Comparing with control group, biomass, exopolysaccharide and ergosterol content of the mature biofilm under isoflavaspidic acid PB (8 MIC) were significantly decreased (P < 0.01). Conclusion Isoflavaspidic acid PB had anti-fungal and fungicidal activities against dermatophytes. Isoflavaspidic acid PB could inhibit the biofilm of Trichophyton. The mechanism might be related to the decline of the biofilm biomass, exopolysaccharide and ergosterol content. These results showed that isoflavaspidic acid PB could be explored for promising anti-biofilm drugs. Graphical abstract

Amelioration of dry eye syndrome in db/db mice with diabetes mellitus by treatment with Tibetan Medicine Formula Jikan Mingmu Drops Publication date: 15 September 2019 Source: Journal of Ethnopharmacology, Volume 241 Author(s): Xiaopeng Ai, Ya Hou, Xiaobo Wang, Xiaoyan Wang, Yusheng Liang, Zhengwen Zhu, Ping Wang, Yong Zeng, Xianjia Li, Xianrong Lai, Xianli Meng, Qi'en Li Abstract Ethnopharmacological relevance Jikan Mingmu Drops (JMD), a traditional Tibetan medicine containing six herbs, has been used to treat dry eye syndrome (DES) in individuals with diabetes mellitus. Aim of study However, the activity of JMD ameliorates DES with diabetes mellitus has not been previously examined. The aim of the study is to investigate the molecular mechanism of JMD on db/db mice. Materials and methods The main chemical constituents of JMD were analyzed by high-performance liquid chromatography and gas chromatography–mass spectrometry. DES was then induced in db/db mice by applying 0.2% benzalkonium chloride to the ocular surface for 7 days. Eye drops containing JMD (0.25, 0.5, or 1 g/mL) or vehicle subsequently were administered three times daily for another 7 days, and the therapeutic effects were evaluated by phenol red thread tear and sodium fluorescein tests. Conjunctival specimens were subjected to hematoxylin and eosin staining and periodic acid–Schiff staining to examine pathological changes and number of goblet cells. ELISA was performed to assess the levels of various inflammatory cytokines. Results JMD contains hydroxysafflor yellow A, magnoflorine, jatrorrhizine hydrochloride, palmatine hydrochloride, berberine hydrochloride, gallic acid, ellagic acid, tauroursodeoxycholic acid, camphor, isoborneol, borneol, trans-cinnamic acid, and muscone. JMD treatment significantly increased the tear volume, decreased the corneal fluorescein staining score, restored the morphology and structure of conjunctival epithelial cells, and markedly downregulated the levels of interleukin (IL)-6, IL-17α, IL-1β, tumor necrosis factor-α, and vascular endothelial growth factor in the conjunctiva. Further data showed that these protective effects were accompanied by inhibition of inflammation in a dose-dependent manner. Conclusions Amelioration of DES in db/db mice with diabetes mellitus by treatment with Tibetan medicine formula JMD maybe related to its anti-inflammatory effects. Graphical abstract

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Cranio-Maxillofacial Surgery

Analysing chin prominence in relation to the lower lip: The lower lip-chin prominence angle Publication date: Available online 9 June 2019 Source: Journal of Cranio-Maxillofacial Surgery Author(s): Farhad B. Naini, Umberto Garagiola, David Wertheim Abstract The purpose of this investigation is to describe a potentially useful analysis in assessing the required extent of sagittal chin augmentation or set-back, by relating chin prominence to lower lip position using the 'lower lip-chin prominence angle'. The secondary aim was to quantitatively evaluate the influence of this angle on perceived attractiveness and desire for surgery. Having described this angular analysis, a quantitative evaluation was undertaken by incrementally altering the angle on an idealised profile image to create a range of images that were rated on a 7-point Likert scale by a pre-selected group of pre-treatment orthognathic patients, clinicians and laypeople. In treatment planning alterations in chin prominence, an 'ideal' sagittal position with soft tissue pogonion on or just behind a true vertical line through the most prominent point of the lower lip may be used. Chin retrusion or prominence up to an angle of 15 degrees retrusion to -5 degrees prominence is deemed acceptable. Surgery is desired from chin prominence of greater than -15 degrees and retrusions greater than 25 degrees. The greater the retrusion or prominence of the chin from an angle of 0 degrees, the less the perceived attractiveness and the greater the desire for surgical correction.

Early Return to Sport Post Maxillofacial Fracture Injury in the Professional Athlete: A Systematic Review Publication date: Available online 8 June 2019 Source: Journal of Cranio-Maxillofacial Surgery Author(s): Umair Ansari, Eugene Wong, John Arvier, Dylan Hyam, Weber Huang Abstract Introduction To summarize the current literature on return to sport times post-maxillofacial fracture injury in the professional athlete. Materials and Methods A literature search on six databases for articles relating to maxillofacial fractures, professional athletes, and return to sport times. Study design, clinical data, and author recommendations were analysed. Results 17 studies were retrieved. One prospective study returned 17 athletes to competitive rugby union and soccer at 3 weeks post injury without complication. Two large retrospective studies (n=278) returned patients to sport at approximately 7 weeks without complication. 64% (n=7) of patients from case based studies returned to sport at 3 to 14 days, 4 of which utilized protective facemasks. Athletes generally returned to competition earlier for lower grade (3 to 10 days) compared to higher grade contact sport (21 days at least). 2 articles recommended a 3 months recovery period for combat sports. 8 articles supported the utility of protective facemasks. Conclusion Early return to sport (<6 weeks) in the professional athlete post maxillofacial fracture injury is achievable. The optimal clinical approach may be to grade the sport according to its impact forces, discuss an early return with reference to the available literature, the potential utility of facemasks, risks of refracture and its operative implications.

Gingival fibroblasts and medication-related osteonecrosis of the jaw: results by real-time and wound healing in vitro assays Publication date: Available online 8 June 2019 Source: Journal of Cranio-Maxillofacial Surgery Author(s): Anna Yuan, Adelheid Munz, Siegmar Reinert, Sebastian Hoefert Abstract Objective This study investigated the effects of bisphosphonates and denosumab on human gingival fibroblasts (HGFs) that could influence inflammation, wound healing, and angiogenesis in medication-related osteonecrosis of the jaw (MRONJ). Methods A real-time in vitro assay was performed on HGFs with and without the addition of bacterial lipopolysaccharide and a mononuclear cell co-culture to observe the effects of zoledronate, ibandronate, alendronate, clodronate, denosumab, and combinations of zoledronate and denosumab at varied concentrations. A wound healing assay was performed, and gene and protein expression was analyzed for inflammatory, angiogenic, and osteoclastogenic cytokines and mediators including interleukin (IL)-1β, IL-6, tumor necrosis factor alpha (TNFα), IL-8, vascular endothelial growth factor (VEGF), RANKL, and osteoprotegerin. Results Higher concentrations of antiresorptives resulted in impaired wound healing and HGF death, which also occurred without mechanical damage. These effects were increased with bacterial lipopolysaccharide and mononuclear cells. Increased levels of IL-1β, TNFα, IL-8, VEGF, osteoprotegerin, and decreased levels of IL-6 were observed. Conclusions Antiresorptive exposure was associated with HGF death and delayed wound healing, which could be attributed to an elevated inflammatory response and immune dysfunction contributing to MRONJ development. There was no evidence of anti-angiogenic effects. Our experiments present the first results of denosumab with HGFs.

A new method for cranial vault reconstruction: augmented reality in synostotic plagiocephaly surgery Publication date: Available online 4 June 2019 Source: Journal of Cranio-Maxillofacial Surgery Author(s): Wenqing Han, Xianxian Yang, Shuihua Wu, Shuangshi Fan, Xiaojun Chen, Mar Aung Zin, Tianjia Liu, Yan Zhang, Shuo Gu, Gang Chai Summary Purpose Augmented reality (AR) is considered to be a valuable tool in craniofacial surgery for preoperative design, intraoperative navigation, and postoperative assessment. Corrective surgery is necessary synostotic plagiocephaly for functional and aesthetic outcomes. Open calvarial reconstruction is a difficult classic surgical procedure with a high accuracy requirement. The purpose of this study was to introduce an AR system application in synostotic plagiocephaly surgery. Materials and Methods Seven plagiocephaly patients (ages 6 months to 24 months, average 16.7 months) were enrolled. Preoperative design was accomplished based on three-dimensional computed tomography (CT) data for patients with synostotic plagiocephaly. We completed the registration with the predefined markers through an image registration process preoperatively. Then, we overlaid the registration results into the surgical field to assist surgeons intraoperatively. CT scans were performed postoperatively. Intracranial volume was measured to judge the surgical outcomes. We performed a quantitative craniometric analysis between the planning of the reconstruction and post-operative results, and the main evaluation indicator was the intracranial volume asymmetry. Results We successfully applied the AR system in patients undergoing synostotic plagiocephaly, providing real-time navigational images of position and orientation information during open calvarial reconstruction surgery in 7 plagiocephaly patients within a span of 5 years. Good appearances were observed after the surgery. Cranial volume asymmetry was decreased from 27.87% to 16.57%, achieving precise intra-operative goals. No significant differences were found between planning and post-operative results. Conclusions The AR system can be applied to plagiocephaly procedures guiding to obtain reliable and accurate results via a precise osteotomy.

Scaffold implantation in the omentum majus of rabbits for new bone formation Publication date: Available online 3 June 2019 Source: Journal of Cranio-Maxillofacial Surgery Author(s): Falk Birkenfeld, Andre Sengebusch, Chiara Völschow, Björn Möller, Hendrik Naujokat, Jörg Wiltfang Abstract Restoration of the mandible after defects caused by ablative surgery remains challenging. Microvascular free flaps from the scapula, fibula or iliac crest remain the 'gold standard'. A drawback of these methods is donor-side morbidity, availability and the shape of the bone. Former cases have shown that prefabrication of a customized bone flap in the latissimus dorsi muscle may be successful; however, this method is still associated with high donor-side morbidity. Osteogenesis in the omentum majus of rabbits by wrapping the periosteum into it was confirmed recently and is particularly interesting for bone endocultivation. Twelve adult male New Zealand white rabbits were used. In each, two hydroxyapatite blocks were implanted in the greater omentum with autologous bone or autologous bone+rhBMP-2. Bone density measurements were performed by CT scans. Fluorochrome labelling was used for new bone formation detection. The animals were sacrificed at week 10, and the specimens were harvested for histological and histomorphometric analysis. In histological and fluorescence microscopic analysis, new bone formation could be found, as well as new blood vessels and connective tissue. No significant differences were found regarding the histological analysis and bone density measurements between the groups. It could be demonstrated that the omentum majus is a practical way to use one's own body as a bioreactor for prefabrication of tissue-engineered bony constructs. Regarding the influence and exact dose of rhBMP-2, further research is necessary. To establish and improve this method, further large-animal experimental studies are also necessary.

Patient-specific, printed titanium implants for reconstruction of mandibular continuity defects: A systematic review of the evidence Publication date: June 2019 Source: Journal of Cranio-Maxillofacial Surgery, Volume 47, Issue 6 Author(s): Alexander MC. Goodson, Madhav A. Kittur, Peter L. Evans, E. Mark Williams

Three-dimensional orbital wall modeling using paranasal sinus segmentation Publication date: June 2019 Source: Journal of Cranio-Maxillofacial Surgery, Volume 47, Issue 6 Author(s): Hannah Kim, Tae-geun Son, Jeonghwan Lee, Hyeun A. Kim, Hyunchul Cho, Woo Shik Jeong, Jong Woo Choi, Youngjun Kim Abstract Purpose Three-dimensional orbital wall modeling is a time-consuming process because of the presence of pseudoforamina. We developed an automated three-dimensional modeling software to characterize the orbital wall, and evaluated it using data from fracture patients. Methods We first characterized the air and face regions using multiphase segmentation; the sinuses were segmented by applying morphological operations to air regions. Pseudoforamina of the orbital wall were offset with the segmented sinuses. Finally, the three-dimensional facial bone model, with orbital wall, was reconstructed from the segmented images. Results Ten computed tomography data sets were used to evaluate the proposed method. Results were compared with those obtained using the active contour model and manual segmentation. The process took 31.7 ± 8.0 s, which was 30–60 times faster than other methods. The average distances between surfaces obtained with the proposed method and those obtained with manual segmentation (normal side: 0.20 ± 0.06 mm; fractured side: 0.28 ± 0.10 mm) were approximately half those obtained using the active contour model. Conclusions Three-dimensional orbital wall models, which were very similar to the manually segmented models, were archived within 1 min using the developed software, regardless of fracture presence. The proposed method might improve the safety and accuracy of surgical procedures.

Analysis of the accuracy of a novel preformed osteosynthesis plate for the reduction and fixation of zygomaticomaxillary complex fractures Publication date: June 2019 Source: Journal of Cranio-Maxillofacial Surgery, Volume 47, Issue 6 Author(s): Philipp Poxleitner, David Steybe, Britta Bublitz, Stefan Schlager, Marc Anton Fuessinger, Pit Jacob Voss, Rainer Schmelzeisen, Carl-Peter Cornelius, Marc Metzger Abstract Introduction There has been a shift toward surgical treatment of ZMC (zygomaticomaxillary complex) fractures with open reduction and subsequent fixation in the past decades. Anatomically preformed osteosynthesis plates, already used in the field of craniomaxillofacial surgery for the treatment of fractures of the mandible and the orbit, might be a suitable option for ZMC fractures as well. Material and methods A statistical shape model was created from 179 cranial CT scans. Based on this surface model, an anatomically preformed plate for the reduction and fixation of ZMC fractures was developed in 3 sizes (S, M, L). Virtual analysis of the accuracy of the plate was performed on a dataset consisting of 120 CT scans. Results Within a determined tolerance range of 0–1.5 mm, analysis revealed a high accuracy of the plate in 70–87 % of the CT scans. The S-sized plate has the highest overall accuracy, whereas the L-sized plate has highest accuracy at the "base" region which is essential for the placement of the plate. Discussion The newly developed plate can be placed via an intraoral approach and analysis of the plate has confirmed its accuracy to be sufficient to ensure an adequate fracture reduction and fixation. It thus might allow for a less extensive approach and less approaches/incisions necessary overall to reduce and fixate ZMC fractures.

Nearthrosis in true long-standing temporomandibular joint dislocation; a report on pathogenesis and clinical features with review of literature Publication date: June 2019 Source: Journal of Cranio-Maxillofacial Surgery, Volume 47, Issue 6 Author(s): Orhan Güven Abstract The behavior and function of the condyle are not the same in every type of temporomandibular joint (TMJ) dislocation. Acute displacement or dislocation of the condyle is not a rare incident, and the treatment modalities have been well known by physicians for a long time. Chronic dislocation of the condyle is considered a relatively common entity for which treatment may indispensably be surgical intervention. Type of dislocation, duration and the number of episodes are taken into account while constructing the treatment plan. Chronic dislocation that has been left untreated for a long time is a relatively less often encountered type exhibiting difficult clinical conditions for treatment. This type of dislocation is usually seen in elderly patients with poor general health conditions and can be classified as "long-standing dislocation." In this clinical condition, after having some chronic dislocation episodes, the condyle leaves the original fossa, sits in the anterior part of the eminence permanently, creates a new fossa and never gets back to the original place again. Duration is the most important criterion in classifying long-standing dislocations and has a great effect on decisions regarding the type of management. The relevant literature includes few reports, most of which speculate upon "duration," which varies on a large scale ranging from 5 weeks to 33 years. There has been neither an agreement on time span within which long-standing dislocation develops, nor a universally accepted definition for what "long-standing" is indeed meant to be. On the other hand, in some cases, the condition has been named "true long-standing dislocation" due to some permanent pathological changes that the TMJ undergoes, such as pseudoarticulation/nearthrosis/false joint/neo-joint. In this paper, management of true long-standing/permanent dislocation in two patients, whose conditions lasted more than 1 year and resulted in permanent changes in TMJ anatomy, is presented. Due to the poor general health condition of the two elderly patients, they were treated in the most conservative way possible. In one of the patients, eminectomy and head-gear application were used to attain gradual relocation of the condyles. Rehabilitation of masticatory function of the other patient was improved prosthetically.

The relationship between temporomandibular joint effusion and pain in patients with internal derangement Publication date: June 2019 Source: Journal of Cranio-Maxillofacial Surgery, Volume 47, Issue 6 Author(s): Hatice Hosgor Abstract Objectives The purpose of this study was to evaluate the relationship between temporomandibular joint (TMJ) effusion and joint pain in patients with internal derangement based on the Diagnostic Criteria for Temporomandibular Disorders (DC/TMD). Methods A total of 240 TMJs from 120 patients with unilateral painful joints (103 females and 17 males, mean age 29.9 ± 12 years) were evaluated. Clinical assessments were carried out according to the DC/TMD guidelines. Magnetic resonance imaging (MRI) was used to evaluate the degree of effusion in each joint. The radiological and clinical findings were analysed for statistically significant correlations. Results Although the results indicated a statistically significant association between moderate joint effusion and disc displacement (p < 0,05), there was no statistically significant association between moderate effusion and joint pain (p > 0,05). There were, however, statistically significant associations between marked effusion and both disc displacement and joint pain (p < 0,05). Conclusion TMJ effusion is associated with both disc displacement and joint pain: the effusion increased in direct proportion to the severity of pain and disc displacement. The possibility that there are various aetiologies for the condition should also be considered.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Neuroscience

This Week in The Journal

Interferon-{gamma} as a Potential Link between Diabetes Mellitus and Dementia

PTCD1 Is Required for Mitochondrial Oxidative-Phosphorylation: Possible Genetic Association with Alzheimer's Disease In addition to amyloid-β plaques and tau tangles, mitochondrial dysfunction is implicated in the pathology of Alzheimer's disease (AD). Neurons heavily rely on mitochondrial function, and deficits in brain energy metabolism are detected early in AD; however, direct human genetic evidence for mitochondrial involvement in AD pathogenesis is limited. We analyzed whole-exome sequencing data of 4549 AD cases and 3332 age-matched controls and discovered that rare protein altering variants in the gene pentatricopeptide repeat-containing protein 1 (PTCD1) show a trend for enrichment in cases compared with controls. We show here that PTCD1 is required for normal mitochondrial rRNA levels, proper assembly of the mitochondrial ribosome and hence for mitochondrial translation and assembly of the electron transport chain. Loss of PTCD1 function impairs oxidative phosphorylation and forces cells to rely on glycolysis for energy production. Cells expressing the AD-linked variant of PTCD1 fail to sustain energy production under increased metabolic stress. In neurons, reduced PTCD1 expression leads to lower ATP levels and impacts spontaneous synaptic activity. Thus, our study uncovers a possible link between a protein required for mitochondrial function and energy metabolism and AD risk. SIGNIFICANCE STATEMENT Mitochondria are the main source of cellular energy and mitochondrial dysfunction is implicated in the pathology of Alzheimer's disease (AD) and other neurodegenerative disorders. Here, we identify a variant in the gene PTCD1 that is enriched in AD patients and demonstrate that PTCD1 is required for ATP generation through oxidative phosphorylation. PTCD1 regulates the level of 16S rRNA, the backbone of the mitoribosome, and is essential for mitochondrial translation and assembly of the electron transport chain. Cells expressing the AD-associated variant fail to maintain adequate ATP production during metabolic stress, and reduced PTCD1 activity disrupts neuronal energy homeostasis and dampens spontaneous transmission. Our work provides a mechanistic link between a protein required for mitochondrial function and genetic AD risk.

Cell-Type-Specific Regulation of Nucleus Accumbens Synaptic Plasticity and Cocaine Reward Sensitivity by the Circadian Protein, NPAS2 The circadian transcription factor neuronal PAS domain 2 (NPAS2) is linked to psychiatric disorders associated with altered reward sensitivity. The expression of Npas2 is preferentially enriched in the mammalian forebrain, including the nucleus accumbens (NAc), a major neural substrate of motivated and reward behavior. Previously, we demonstrated that downregulation of NPAS2 in the NAc reduces the conditioned behavioral response to cocaine in mice. We also showed that Npas2 is preferentially enriched in dopamine receptor 1 containing medium spiny neurons (D1R-MSNs) of the striatum. To extend these studies, we investigated the impact of NPAS2 disruption on accumbal excitatory synaptic transmission and strength, along with the behavioral sensitivity to cocaine reward in a cell-type-specific manner. Viral-mediated knockdown of Npas2 in the NAc of male and female C57BL/6J mice increased the excitatory drive onto MSNs. Using Drd1a-tdTomato mice in combination with viral knockdown, we determined these synaptic adaptations were specific to D1R-MSNs relative to non-D1R-MSNs. Interestingly, NAc-specific knockdown of Npas2 blocked cocaine-induced enhancement of synaptic strength and glutamatergic transmission specifically onto D1R-MSNs. Last, we designed, validated, and used a novel Cre-inducible short-hairpin RNA virus for MSN-subtype-specific knockdown of Npas2. Cell-type-specific Npas2 knockdown in D1R-MSNs, but not D2R-MSNs, in the NAc reduced cocaine conditioned place preference. Together, our results demonstrate that NPAS2 regulates excitatory synapses of D1R-MSNs in the NAc and cocaine reward-related behavior. SIGNIFICANCE STATEMENT Drug addiction is a widespread public health concern often comorbid with other psychiatric disorders. Disruptions of the circadian clock can predispose or exacerbate substance abuse in vulnerable individuals. We demonstrate a role for the core circadian protein, NPAS2, in mediating glutamatergic neurotransmission at medium spiny neurons (MSNs) in the nucleus accumbens (NAc), a region critical for reward processing. We find that NPAS2 negatively regulates functional excitatory synaptic plasticity in the NAc and is necessary for cocaine-induced plastic changes in MSNs expressing the dopamine 1 receptor (D1R). We further demonstrate disruption of NPAS2 in D1R-MSNs produces augmented cocaine preference. These findings highlight the significance of cell-type-specificity in mechanisms underlying reward regulation by NPAS2 and extend our knowledge of its function.

Sexually Dimorphic Regulation of Behavioral States by Dopamine in Caenorhabditis elegans Sex differences in behavior allow animals to effectively mate and reproduce. However, the mechanism by which biological sex regulates behavioral states, which underlie the regulation of sex-shared behaviors, such as locomotion, is largely unknown. In this study, we studied sex differences in the behavioral states of Caenorhabditis elegans and found that males spend less time in a low locomotor activity state than hermaphrodites and that dopamine generates this sex difference. In males, dopamine reduces the low activity state by acting in the same pathway as polycystic kidney disease-related genes that function in male-specific neurons. In hermaphrodites, dopamine increases the low activity state by suppression of octopamine signaling in the sex-shared SIA neurons, which have reduced responsiveness to octopamine in males. Furthermore, dopamine promotes exploration both inside and outside of bacterial lawn (the food source) in males and suppresses it in hermaphrodites. These results demonstrate that sexually dimorphic signaling allows the same neuromodulator to promote adaptive behavior for each sex. SIGNIFICANCE STATEMENT The mechanisms that generate sex differences in sex-shared behaviors, including locomotion, are not well understood. We show that there are sex differences in the regulation of behavioral states in the model animal Caenorhabditis elegans. Dopamine promotes the high locomotor activity state in males, which must search for mates to reproduce, and suppresses it in self-fertilizing hermaphrodites through distinct molecular mechanisms. This study demonstrates that sex-specific signaling generates sex differences in the regulation of behavioral states, which in turn modulates the locomotor activity to suit reproduction for each sex.

Reorganization of Recurrent Layer 5 Corticospinal Networks Following Adult Motor Training Recurrent synaptic connections between neighboring neurons are a key feature of mammalian cortex, accounting for the vast majority of cortical inputs. Although computational models indicate that reorganization of recurrent connectivity is a primary driver of experience-dependent cortical tuning, the true biological features of recurrent network plasticity are not well identified. Indeed, whether rewiring of connections between cortical neurons occurs during behavioral training, as is widely predicted, remains unknown. Here, we probe M1 recurrent circuits following motor training in adult male rats and find robust synaptic reorganization among functionally related layer 5 neurons, resulting in a 2.5-fold increase in recurrent connection probability. This reorganization is specific to the neuronal subpopulation most relevant for executing the trained motor skill, and behavioral performance was impaired following targeted molecular inhibition of this subpopulation. In contrast, recurrent connectivity is unaffected among neighboring layer 5 neurons largely unrelated to the trained behavior. Training-related corticospinal cells also express increased excitability following training. These findings establish the presence of selective modifications in recurrent cortical networks in adulthood following training. SIGNIFICANCE STATEMENT Recurrent synaptic connections between neighboring neurons are characteristic of cortical architecture, and modifications to these circuits are thought to underlie in part learning in the adult brain. We now show that there are robust changes in recurrent connections in the rat motor cortex upon training on a novel motor task. Motor training results in a 2.5-fold increase in recurrent connectivity, but only within the neuronal subpopulation most relevant for executing the new motor behavior; recurrent connectivity is unaffected among adjoining neurons that do not execute the trained behavior. These findings demonstrate selective reorganization of recurrent synaptic connections in the adult neocortex following novel motor experience, and illuminate fundamental properties of cortical function and plasticity.

Regeneration of Dopaminergic Neurons in Adult Zebrafish Depends on Immune System Activation and Differs for Distinct Populations Adult zebrafish, in contrast to mammals, regenerate neurons in their brain, but the extent and variability of this capacity is unclear. Here we ask whether the loss of various dopaminergic neuron populations is sufficient to trigger their functional regeneration. Both sexes of zebrafish were analyzed. Genetic lineage tracing shows that specific diencephalic ependymo-radial glial (ERG) progenitor cells give rise to new dopaminergic [tyrosine hydroxylase-positive (TH+)] neurons. Ablation elicits an immune response, increased proliferation of ERG progenitor cells, and increased addition of new TH+ neurons in populations that constitutively add new neurons (e.g., diencephalic population 5/6). Inhibiting the immune response attenuates neurogenesis to control levels. Boosting the immune response enhances ERG proliferation, but not addition of TH+ neurons. In contrast, in populations in which constitutive neurogenesis is undetectable (e.g., the posterior tuberculum and locus ceruleus), cell replacement and tissue integration are incomplete and transient. This is associated with a loss of spinal TH+ axons, as well as permanent deficits in shoaling and reproductive behavior. Hence, dopaminergic neuron populations in the adult zebrafish brain show vast differences in regenerative capacity that correlate with constitutive addition of neurons and depend on immune system activation. SIGNIFICANCE STATEMENT Despite the fact that zebrafish show a high propensity to regenerate neurons in the brain, this study reveals that not all types of dopaminergic neurons are functionally regenerated after specific ablation. Hence, in the same adult vertebrate brain, mechanisms of successful and incomplete regeneration can be studied. We identify progenitor cells for dopaminergic neurons and show that activating the immune system promotes the proliferation of these cells. However, in some areas of the brain this only leads to insufficient replacement of functionally important dopaminergic neurons that later disappear. Understanding the mechanisms of regeneration in zebrafish may inform interventions targeting the regeneration of functionally important neurons, such as dopaminergic neurons, from endogenous progenitor cells in nonregenerating mammals.

The Spinal Transcriptome after Cortical Stroke: In Search of Molecular Factors Regulating Spontaneous Recovery in the Spinal Cord In response to cortical stroke and unilateral corticospinal tract degeneration, compensatory sprouting of spared corticospinal fibers is associated with recovery of skilled movement in rodents. To date, little is known about the molecular mechanisms orchestrating this spontaneous rewiring. In this study, we provide insights into the molecular changes in the spinal cord tissue after large ischemic cortical injury in adult female mice, with a focus on factors that might influence the reinnervation process by contralesional corticospinal neurons. We mapped the area of cervical gray matter reinnervation by sprouting contralesional corticospinal axons after unilateral photothrombotic stroke of the motor cortex in mice using anterograde tracing. The mRNA profile of this reinnervation area was analyzed using whole-genome sequencing to identify differentially expressed genes at selected time points during the recovery process. Bioinformatic analysis revealed two phases of processes: early after stroke (4–7 d post-injury), the spinal transcriptome is characterized by inflammatory processes, including phagocytic processes as well as complement cascade activation. Microglia are specifically activated in the denervated corticospinal projection fields in this early phase. In a later phase (28–42 d post-injury), biological processes include tissue repair pathways with upregulated genes related to neurite outgrowth. Thus, the stroke-denervated spinal gray matter, in particular its intermediate laminae, represents a growth-promoting environment for sprouting corticospinal fibers originating from the contralesional motor cortex. This dataset provides a solid starting point for future studies addressing key elements of the post-stroke recovery process, with the goal to improve neuroregenerative treatment options for stroke patients. SIGNIFICANCE STATEMENT We show that the molecular changes in the spinal cord target tissue of the stroke-affected corticospinal tract are mainly defined by two phases: an early inflammatory phase during which microglia are specifically activated in the target area of reinnervating corticospinal motor neurons; and a late phase during which growth-promoting factors are upregulated which can influence the sprouting response, arborization, and synapse formation. By defining for the first time the endogenous molecular machinery in the stroke-denervated cervical spinal gray matter with a focus on promotors of axon growth through the growth-inhibitory adult CNS, this study will serve as a basis to address novel neuroregenerative treatment options for chronic stroke patients.

Pedunculopontine Glutamatergic Neurons Provide a Novel Source of Feedforward Inhibition in the Striatum by Selectively Targeting Interneurons The main excitatory inputs to the striatum arising from the cortex and the thalamus innervate both striatal spiny projection neurons and interneurons. These glutamatergic inputs to striatal GABAergic interneurons have been suggested to regulate the spike timing of striatal projection neurons via feedforward inhibition. Understanding how different excitatory inputs are integrated within the striatal circuitry and how they regulate striatal output is crucial for understanding basal ganglia function and related behaviors. Here, using VGLUT2 mice from both sexes, we report the existence of a glutamatergic projection from the mesencephalic locomotor region to the striatum that avoids the spiny neurons and selectively innervates interneurons. Specifically, optogenetic activation of glutamatergic axons from the pedunculopontine nucleus induced monosynaptic excitation in most recorded striatal cholinergic interneurons and GABAergic fast-spiking interneurons. Optogenetic stimulation in awake head-fixed mice consistently induced an increase in the firing rate of putative cholinergic interneurons and fast-spiking interneurons. In contrast, this stimulation did not induce excitatory responses in spiny neurons but rather disynaptic inhibitory responses ex vivo and a decrease in their firing rate in vivo, suggesting a feedforward mechanism mediating the inhibition of spiny projection neurons through the selective activation of striatal interneurons. Furthermore, unilateral stimulation of pedunculopontine nucleus glutamatergic axons in the striatum induced ipsilateral head rotations consistent with the inhibition of striatal output neurons. Our results demonstrate the existence of a unique interneuron-specific midbrain glutamatergic input to the striatum that exclusively recruits feedforward inhibition mechanisms. SIGNIFICANCE STATEMENT Glutamatergic inputs to the striatum have been shown to target both striatal projection neurons and interneurons and have been proposed to regulate spike timing of the projection neurons in part through feedforward inhibition. Here, we reveal the existence of a midbrain source of glutamatergic innervation to the striatum, originating in the pedunculopontine nucleus. Remarkably, this novel input selectively targets striatal interneurons, avoiding the projection neurons. Furthermore, we show that this selective innervation of interneurons can regulate the firing of the spiny projection neurons and inhibit the striatal output via feedforward inhibition. Together, our results describe a unique source of excitatory innervation to the striatum which selectively recruits feedforward inhibition of spiny neurons without any accompanying excitation.

Single-Cell Membrane Potential Fluctuations Evince Network Scale-Freeness and Quasicriticality What information single neurons receive about general neural circuit activity is a fundamental question for neuroscience. Somatic membrane potential (Vm) fluctuations are driven by the convergence of synaptic inputs from a diverse cross-section of upstream neurons. Furthermore, neural activity is often scale-free, implying that some measurements should be the same, whether taken at large or small scales. Together, convergence and scale-freeness support the hypothesis that single Vm recordings carry useful information about high-dimensional cortical activity. Conveniently, the theory of "critical branching networks" (one purported explanation for scale-freeness) provides testable predictions about scale-free measurements that are readily applied to Vm fluctuations. To investigate, we obtained whole-cell current-clamp recordings of pyramidal neurons in visual cortex of turtles with unknown genders. We isolated fluctuations in Vm below the firing threshold and analyzed them by adapting the definition of "neuronal avalanches" (i.e., spurts of population spiking). The Vm fluctuations which we analyzed were scale-free and consistent with critical branching. These findings recapitulated results from large-scale cortical population data obtained separately in complementary experiments using microelectrode arrays described previously (Shew et al., 2015). Simultaneously recorded single-unit local field potential did not provide a good match, demonstrating the specific utility of Vm. Modeling shows that estimation of dynamical network properties from neuronal inputs is most accurate when networks are structured as critical branching networks. In conclusion, these findings extend evidence of critical phenomena while also establishing subthreshold pyramidal neuron Vm fluctuations as an informative gauge of high-dimensional cortical population activity. SIGNIFICANCE STATEMENT The relationship between membrane potential (Vm) dynamics of single neurons and population dynamics is indispensable to understanding cortical circuits. Just as important to the biophysics of computation are emergent properties such as scale-freeness, where critical branching networks offer insight. This report makes progress on both fronts by comparing statistics from single-neuron whole-cell recordings with population statistics obtained with microelectrode arrays. Not only are fluctuations of somatic Vm scale-free, they match fluctuations of population activity. Thus, our results demonstrate appropriation of the brain's own subsampling method (convergence of synaptic inputs) while extending the range of fundamental evidence for critical phenomena in neural systems from the previously observed mesoscale (fMRI, LFP, population spiking) to the microscale, namely, Vm fluctuations.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com