AAV-mediated gene delivery of the calreticulin anti-angiogenic domain inhibits ocular neovascularization

Abstract

Ocular neovascularization is a common pathological feature in diabetic retinopathy and neovascular age-related macular degeneration that can lead to severe vision loss. We evaluated the therapeutic efficacy of a novel endogenous inhibitor of angiogenesis, the calreticulin anti-angiogenic domain (CAD180), and its functional 112-residue fragment, CAD-like peptide 112 (CAD112), delivered using a self-complementary adeno-associated virus serotype 2 (scAAV2) in rodent models of oxygen-induced retinopathy and laser-induced choroidal neovascularization. The expression of CAD180 and CAD112 was elevated in human umbilical vein endothelial cells transduced with scAAV2-CAD180 or scAAV2-CAD112, respectively, and both inhibited angiogenic activity in vitro. Intravitreal gene delivery of scAAV2-CAD180 or scAAV2-CAD112 significantly inhibited ischemia-induced retinal neovascularization in rat eyes (CAD180: 52.7% reduction; CAD112: 49.2% reduction) compared to scAAV2-mCherry, as measured in retinal flatmounts stained with isolectin B4. Moreover, the retinal structure and function were unaffected by scAAV2-CAD180 or scAAV2-CAD112, as measured by optical coherence tomography and electroretinography. Moreover, subretinal delivery of scAAV2-CAD180 or scAAV2-CAD112 significantly attenuated laser-induced choroidal neovascularization in mouse eyes compared to scAAV2-mCherry, as measured by fundus fluorescein angiography (CAD180: 62.4% reduction; CAD112: 57.5% reduction) and choroidal flatmounts (CAD180: 40.21% reduction; CAD112: 43.03% reduction). Gene delivery using scAAV2-CAD180 or scAAV2-CAD112 has significant potential as a therapeutic option for the management of ocular neovascularization.

Response to Comment on Mycobacterium chelonae Infection of the Buttocks Secondary to Lipofilling: A Case Report and Review of the Literature

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Prolapsed bladder following rupture of patent urachal cyst, mimicking bladder exstrophy: a case report and literature review

Abstract

A 26-year-old pregnant woman was diagnosed with fetal bladder prolapse following rupture of a patent urachus/urachal cyst, based on the finding of cyst disappearance with replacement with an infra-umbilical, extra-abdominal solid soft-tissue mass, mimicking bladder exstrophy. The neonatal findings confirmed the prenatal diagnosis. The baby was healthy and had a successful surgical correction. This report provides clues to differentiating ruptured bladder prolapse from bladder exstrophy as follows: (1) well-documented urachal cyst with urine-filled mass in the early gestation, (2) development of solid soft-tissue mass shortly after disappearance of the urachal cyst, and (3) no other structural abnormalities (bladder exstrophy is usually associated with abnormal genitalia, epispadias, or pubic diastasis). This study underlines the differentiation between the two entities because of the vast difference in prognosis, management, and proper counseling.

Minimally invasive esophagectomy in the prone position improves postoperative outcomes: role of C-reactive protein as an indicator of surgical invasiveness

Abstract

Background

The aim of the study was to assess serum C-reactive protein (CRP) level immediately after minimally invasive esophagectomy (MIE) as a surrogate of surgical invasiveness in patients who underwent esophagectomy.

Methods

In total, 104 patients were enrolled in the study: 37 patients underwent MIE in the left lateral decubitus position (MIE-LP) and 67 patients underwent MIE in the prone position (MIE-PP). Serum CRP levels were assessed on POD 1, 3, 5, and 7 after MIE, and were compared with surgical outcomes and duration of systemic inflammatory response syndrome (SIRS) to investigate less invasiveness of the MIE.

Results

Reduced serum CRP level on POD 1 was associated with PP during MIE (P < 0.001) and decreased blood loss (P = 0.03). MIE-PP was identified as a significant independent predictor of reduced CRP level on POD 1 (odds ratio 3.65, P = 0.042). CRP level on POD 7 was associated with gender (P = 0.02), position of MIE (P = 0.011), blood loss (P = 0.02), and respiratory complications and/or anastomotic leakage (P < 0.001). Postoperative respiratory and/or anastomotic complication was identified as a significant predictor of elevated serum CRP level on POD 7 (odds ratio 3.44, P = 0.048). Shorter duration of SIRS was shown in the patients with reduced serum CRP level on POD 1 and 7 (P = 0.03 and P < 0.001, respectively).

Conclusion

Serial assessments of serum CRP level immediately after MIE may be a possible indicator that can reflect surgical invasiveness and postoperative complications.

The exposure to water with cigarette residue changes the anti-predator response in female Swiss albino mice

Abstract

Recent studies have shown that cigarette consumption affects much more than human health. Smoked cigarette butt (SCB) disposal into the environment can bring little-known negative biological consequences to mammals, since it contains many organic and inorganic toxic chemical constituents. Thus, we aim at assessing whether the ingestion of water with leached SCB for 60 days by female Swiss mice changes their defensive behavioral response to potential predators (cats and snakes). We worked with the following groups of animals: control (pollutant-free water), water with environmental concentration of SCB (1.9 μg/L of nicotine), and concentration 1000 times higher (EC1000×). Our data show that the treatments did not cause locomotor, visual, auditory, and olfactory deficit in the animals. However, we observed that the animals exposed to the pollutants did not present behavioral differences in the test session with or without the snake. On the other hand, animals in all groups showed defensive behavior when the test was conducted with the cat in the apparatus. However, female mice presented weaker response than the control. Thus, our data point towards the potential neurotoxic damage caused to mice who have ingested water with SCB residues, even at low concentrations.

Acquired idiopathic generalized anhidriosis: Successful treatment with cyclosporine in two cases

Transcriptomic profiling of interacting nasal staphylococci species reveals global changes in gene and non-coding RNA expression

Abstract

Interspecies interactions between bacterial pathogens and the commensal microbiota can influence disease outcome. In the nasal cavities, Staphylococcus epidermidis has been shown to be a determining factor for Staphylococcus aureus colonization and biofilm formation. However, the interaction between S. epidermidis and S. aureus has mainly been described by phenotypic analysis, and little is known about how this interaction modulates gene expression.This study aimed to determine the interactome of nasal S. aureus and S. epidermidis isolates to understand the molecular effect of interaction. After whole-genome sequencing of two nasal staphylococcal isolates, an agar-based RNA sequencing setup was utilized to identify interaction-induced transcriptional alterations in surface-associated populations. Our results revealed differential expression of several virulence genes in both species. We also identified putative non-coding RNAs (ncRNAs) and, interestingly, detected a putative ncRNA transcribed antisense to esp, the serine protease of S. epidermidis, that has previously been shown to inhibit nasal colonization of S. aureus. In our study, the gene encoding Esp and the antisense ncRNA are both downregulated during interaction with S. aureus. Our findings contribute to a better understanding of pathogen physiology in the context of interactions with the commensal microbiota, and may provide targets for future therapeutics.

Amplicon-based next-generation sequencing of plasma cell-free DNA for detection of driver and resistance mutations in advanced non-small cell lung cancer

Abstract

Background

Genomic analysis of plasma cell-free DNA is transforming lung cancer care, however available assays are limited by cost, turnaround time, and imperfect accuracy. Here we study amplicon-based plasma next-generation sequencing (NGS), rather than hybrid-capture-based plasma NGS, hypothesizing this would allow sensitive detection and monitoring of driver and resistance mutations in advanced non-small cell lung cancer (NSCLC).

Methods

Plasma samples from patients with NSCLC and a known targetable genotype (EGFR, ALK/ROS1 and other rare genotypes) were collected while on therapy and analyzed, blinded to tumor genotype. Plasma NGS was performed using enhanced tagged amplicon sequencing of hotspots and coding regions from 36 genes, as well as intronic coverage for detection of ALK/ROS1 fusions. Diagnostic accuracy was compared to plasma ddPCR and tumor genotype.

Results

A total of 168 specimens from 46 patients were studied. Matched plasma NGS and ddPCR across 120 variants from 80 samples revealed high concordance of allelic fraction (R²=0.95). Pretreatment, sensitivity of plasma NGS for the detection of EGFR driver mutations was 100% (30/30), compared to 87% for ddPCR (26/30). A full spectrum of rare driver oncogenic mutations could be detected including sensitive detection of ALK/ROS1 fusions (8/9 detected, 89%). Studying 25 patients positive for EGFR T790M that developed resistance to osimertinib, 15 resistance mechanisms could be detected including tertiary EGFR mutations (C797S, Q791P) and mutations or amplifications of non-EGFR genes, some of which could be detected pre-treatment or months before progression.

Conclusions

This blinded analysis demonstrates the ability of amplicon-based plasma NGS to detect a full range of targetable genotypes in NSCLC, including fusion genes, with high accuracy. The ability of plasma NGS to detect a range of pre-existing and acquired resistance mechanisms highlights its potential value as an alternative to single mutation digital PCR-based plasma assays for personalizing treatment of TKI resistance in lung cancer.

Prognostic and predictive role of neutrophil/lymphocytes ratio in metastatic colorectal cancer: a retrospective analysis of the TRIBE study by GONO.

Abstract

Background

Neutrophil/Lymphocyte ratio (NLR), defined as absolute neutrophils count divided by absolute lymphocytes count, has been reported as poor prognostic factor in several neoplastic diseases but only a few data are available about unresectable metastatic colorectal cancer (mCRC) patients (pts). The aim of our study was to evaluate the prognostic and predictive role of NLR in the TRIBE trial.

Patients and Methods

Pts enrolled in TRIBE trial were included. TRIBE is a multicentre phase III trial randomizing unresectable and previously untreated mCRC pts to receive FOLFOXIRI or FOLFIRI plus bevacizumab. A cut-off value of 3 was adopted to discriminate pts with low (NLR < 3) vs high (NLR ≥ 3) NLR, as primary analysis. As secondary analysis, NLR was treated as an ordinal variable with three levels based on terciles distribution.

Results

NLR at baseline was available for 413 patients. After multiple imputation at univariate analysis, patients with high NLR had significantly shorter PFS (HR: 1.27 [95%CI:1.05-1.55], p=0.017) and OS (HR: 1.56 [95%CI: 1.25-1.95], p<0.001) than patients with low NLR. In the multivariable model, NLR retained a significant association with OS (HR: 1.44 (95%CI: 1.14-1.82), p=0.014) but not with PFS (HR: 1.18 [95%CI: 0.95-1.46], p=0.375). No interaction effect between treatment arm and NLR was evident in terms of PFS (p for interaction=0.536) or OS (p for interaction=0.831). Patients with low (HR: 0.84 [95%CI: 0.64-1.08]) and high (HR: 0.73 [0.54-0.97]) NLR achieved similar PFS benefit from the triplet and consistent results were obtained in terms of OS (HR: 0.83 [95%CI: 0.62-1.12] for low NLR; HR: 0.82 [95%CI: 0.59-1.12] for high NLR).

Conclusion

This study confirmed the prognostic role of NLR in mCRC pts treated with bevacizumab plus chemotherapy in first line, showing the worse prognosis of pts with high NLR. The advantage of the triplet is independent of NLR at baseline.

Investigation of the effectiveness of therapeutic ultrasound with high-resolution ultrasonographic cross-sectional area measurement of cervical nerve roots in patients with chronic cervical radiculopathy: a prospective, controlled, single-blind study

Abstract

Purpose

The purpose of our study was to evaluate the effects of therapeutic ultrasound (US) on chronic cervical radiculopathy (CR) patients using both the clinical parameters and the cross-sectional area (CSA) values of the cervical nerve roots (NR) measured by high-resolution ultrasonography (HRUS).

Methods

Thirty-two patients with chronic CR were included in this prospective, controlled, and single-blind study. All of the patients received therapeutic US at continuous mode, 1-MHz frequency, and 1.5-W/cm² intensity for 10 sessions. The patients were assessed using VAS for pain, Modified Neck Disability Index, and Short-form 12, and cervical NR were examined with HRUS. The data were obtained before treatment (W0), the second week immediately after treatment (W2), and at the sixth week (W6).

Results

Twenty-nine patients and a total of 42 affected cervical NR and 42 unaffected cervical NR (control group) were evaluated. A significant improvement was observed for all clinical parameters and CSA values of affected cervical NR both at W2 and W6 compared to pre-treatment values (p < 0.05).

Conclusions

We found therapeutic US to be beneficial in improvement of pain, disability, and quality of life of patients with chronic CR. We suggest that CSA measurements may also contribute to both diagnosis and post-treatment evaluation in chronic CR.

Neuroendokrine Tumoren aus psychoonkologischer Sicht – ein Überblick

Zusammenfassung

In aller Regel besteht bei Patienten mit neuroendokrinen Tumoren (NET) ein langwieriger Verlauf, der bereits lange vor Diagnosestellung beginnt. Sie müssen mit viel Ungewissheit umgehen. Dennoch lassen sich allgemeine Aussagen zur Lebensqualität (LQ) und psychischen Verfassung von Patienten mit NET schwer treffen. Zum einen ist NET ein Oberbegriff für eine heterogene Gruppe von Tumoren, die sich in ihrem klinischen Erscheinungsbild stark unterscheiden. Somit gibt es zwangsläufig innerhalb der Gruppe von Patienten mit NET erhebliche Unterschiede in den Belastungen und der LQ. Zum anderen gibt es bislang in der Literatur nur wenige hochwertige Studien, die mittels validierter und NET-spezifischer Messverfahren die LQ bzw. Belastungen von Patienten mit NET analysieren. Um besonders belastete Patienten zu identifizieren, ist eine differenzierte Betrachtung der LQ von Patienten mit NET erforderlich. Nur dann lassen sich auch klinisch relevante Schlüsse ableiten. Für Patienten mit psychischen Belastungen gibt es verschiedene evidenzbasierte Behandlungsmöglichkeiten.

Psychosoziale Aspekte von Harnblasenkarzinompatienten

Zusammenfassung

Tumorpatienten müssen sich mit dem Gefühl der Unsicherheit hinsichtlich des weiteren Lebens und Krankheitsverlaufs sowie mit unerwünschten Nebenwirkungen der Therapie auseinandersetzen. Niedrige Lebensqualität ist bei krebskranken Patienten ein unabhängiger prognostischer Faktor für geringere Überlebensraten. Psychosozialer Stress beeinflusst negativ das Immunsystem (Reduktion der NK-Zellen, Anstieg des Cortisolspiegels). Trotz verbesserter Diagnostik und Therapie bleibt das Blasenkarzinom durch hohe Rezidiv- und Progressionshäufigkeit gekennzeichnet. Für die Behandlung eines Harnblasentumors werden, je nach Tumorstadium, verschiedene therapeutische Maßnahmen empfohlen, die sich auf die Lebensqualität auswirken können. Studien über die psychosozialen Bedürfnisse von Patienten mit Harnblasenkarzinom sind vergleichsweise selten.

Physics’ silence on time

Abstract

In this paper I argue that physics is, always was, and probably always will be voiceless with respect to tense and passage, and that, therefore, if, as I believe, tense and passage are the essence of time, physics' contribution to our understanding of time can only be limited. The argument, in a nutshell, is that if "physics has no possibility of expression for the Now", to quote Einstein, then it cannot add anything to the study of tense and passage, and specifically, cannot add anything to the debate between deniers and affirmers of the existence or reality of tense and passage. Since relativity theory did not equip physics with a new language with which to speak of tense and passage, I draw the further conclusion that relativity theory has not generated the revolution to our conception of time that is attributed to it. In the last section I discuss the motivations behind the continued but misguided attempts to integrate tense into a relativistic setting, and assess the manners in which relativity theory has nevertheless enhanced, albeit indirectly, our understanding of tense and passage.

Etoposide as a single agent in the treatment of mycosis fungoides: A retrospective analysis

Abstract

Several chemotherapy agents have shown efficacy in the treatment of mycosis fungoides (MF). In the literature, there is limited data on the use of single agent etoposide for MF. We aimed to retrospectively review our experience with single agent etoposide in the treatment of advanced-stage or refractory early-stage MF with focus on analyzing its efficacy and safety. We included 13 MF patients who were treated with single agent etoposide. Patients were identified through the Cutaneous T Cell Lymphoma Database of Indiana University that involves patients treated from 2006 to 2016. Overall nine patients (69%) responded to treatment. No complete response was identified. Median time to response was 12.5 weeks (range: 6–25.4). Median duration of response was 43 weeks (range: 5–60) and median time to treatment failure was 31.3 weeks (range: 12.4–230). Hematological toxicity was observed in eight patients including two patients with grade 4 neutropenia and/or lymphopenia leading to sepsis. Higher doses of etoposide were significantly correlated with higher grades of anemia, neutropenia or lymphopenia (p < .05). Our study demonstrates that etoposide is an effective treatment for MF and may be considered in selected patients with progressive MF who have failed other treatments.

Correction to: Possessing epistemic reasons: the role of rational capacities

Abstract

In the original publication of the article, the last sentence in footnote 16 was incorrectly published as "Thanks to—for raising this issue." The corrected sentence should read as "Thanks to Daniel Star for raising this issue."

The Stress Factor of Social Media

The Pros and Cons of the PROs

Speckled lentiginous nevus: understanding the process of development and regression

Speckled lentiginous nevus (SLN) is characterized by a brownish macule studded with blackish, multiple melanocytic nevi, although the cutaneous manifestations may vary by cases, and change by age. Therefore, there has been disagreement as to whether SLN is a congenital or acquired disease 1, 2). In rare cases of SLN, halo phenomenon may occur around the pre-existing lesion, associated with the coexistence of distant vitiligo 3). Here, we report a patient whose café-au-lait spot at birth changed to SLN, followed by the occurrence of vitiligo and poliosis through the process of spontaneous regression phenomenon against the SLN. We examined T-cell clones infiltrating around the melanocytic nevi and in the patient's peripheral blood.

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