Immunotherapy With Low-Dose IL-2/CD25 Prevents ß-Cell Dysfunction and Dysglycemia in Prediabetic NOD Mice.

Low-dose IL-2 is a promising immunotherapy in clinical trials for treating type 1 diabetes. A new IL-2 analog, IL-2/CD25 fusion protein, has been shown to more efficiently delay or prevent diabetes in NOD mice by expanding the population of activated regulatory T cells. This therapy is intended for use before clinical diagnosis, in the early stages of type 1 diabetes progression. During this prediabetic period, there is a chronic decline in ß-cell function that has long-term implications for disease pathogenesis. Yet, to date, the effects of IL-2/CD25 on ß-cell function have not been evaluated. In this study, we treated prediabetic NOD mice with low-dose mouse IL-2/CD25 over 5 weeks and determined its impact on ß-cell function. This treatment limited the progressive impairment of glucose tolerance and insulin secretion typical of the later stages of prediabetes. Intracellular Ca2+ responses to glucose in ß-cells became more robust and synchronous, indicating that changing the local immune cell infiltrate with IL-2/CD25 preserved ß-cell function even after treatment cessation. Our study thus provides mechanistic insight and serves as a steppingstone for future research using low-dose IL-2/CD25 immunotherapy in patients. ARTICLE HIGHLIGHTS: Immunotherapies such as IL-2/CD25 are known to prevent or delay diabetes. However, their impact on individual ß-cell function is not yet understood. Female NOD mice progress from stage 1 to 2 pre-type 1 diabetes between 12 and 17 weeks. Treatment with mouse IL-2 (mIL-2)/CD25 prevents this progression even after treatment cessation. Individual ß-cell function (measured via intracellular Ca2+ responses to glucose) declines during the pathogenesis of type 1 diabetes. Treatment with mIL-2/CD25 therapy limits ß-cell dysfunction, and function continues to improve after treatment cessation. Insulin secretion is improved with mIL-2/CD25 therapy.

An Update on the Use of Wearable Devices in Men's Health.

Men's health represents an often-overlooked aspect of public health. Men have higher mortality rates worldwide and are more negatively affected by chronic conditions such as obesity and heart disease, as well as addiction to alcohol and tobacco. Men also have health issues such as prostate cancer and male sexual dysfunction which only affect them. Because of the skewed burden of morbidity and mortality on men, it is imperative from a public health perspective to make a concerted effort to specifically improve men's health. The use of wearable devices in medical practice presents a novel avenue to invest in men's health in a safe, easily scalable, and economic fashion. Wearable devices are now ubiquitous in society, and their use in the healthcare setting is only increasing with time. There are commercially available devices such as smart watches which are available to lay people and healthcare professionals alike to improve overall health and wellness, and there are also purpose-built wearable devices which are used to track or treat a specific disease. In our review of the literature, we found that while research in the field of wearable devices is still in its early stages, there is ample evidence that wearable devices can greatly improve men's health in the long-term.

Magnitude and pattern of anxiety levels with gender wise predilection of coping strategies amid resident doctors of emergency department.

Objectives: To screen and assess the severity level of anxiety, its influencing factors along with the gender-wise predilection of coping strategies among resident doctors working in accident and emergency departments. Methods: A transverse study was conducted amongst 260 resident doctors of accident and emergency department of different teaching hospitals of Karachi from October 2020 until March 2021. A demographic sheet containing questions related to factors, GAD-7 (Generalized Anxiety Disorder) and Brief COPE were used to measure the severity level of anxiety and coping strategies. Data was scored according to the standard scoring procedure for each subscale and for further statistical analysis SPSS Version 21 was used. Results: Out of all participants, 68.1% were <30 years of age, 63.1% were females while 54.2% were single. The findings of the study showed the prevalence of anxiety within the range of normal (38.1%), mild (35.0%), moderate (16.9%) and severe (10.0%). Gender (p= 0.001), marital status (p= 0.040) and job satisfaction (p= 0.007) among resident doctors were significantly associated with level of anxiety. Deemed to coping strategies, the most frequently were planning (n=145, 90.0%), acceptance (n=141, 87.6%), and religion (n=137, 85.1 %). All coping strategies were mostly opted by females except substance abuse. Conclusion: More than a half of the resident doctors manifested with mild to severe anxiety disorder, which highlights the need for psychological support programs to minimize anxiety levels and to ensure a healthy environment at workplace for the health practitioners.

Derivation and Characterization of Murine and Amphibian Müller Glia Cell Lines.

Purpose: Müller glia (MG) in the retina of Xenopus laevis (African clawed frog) reprogram to a transiently amplifying retinal progenitor state after an injury. These progenitors then give rise to new retinal neurons. In contrast, mammalian MG have a restricted neurogenic capacity and undergo reactive gliosis after injury. This study sought to establish MG cell lines from the regeneration-competent frog and the regeneration-deficient mouse. Methods: MG were isolated from postnatal day 5 GLAST-CreERT; Rbfl/fl mice and from adult (3-5 years post-metamorphic) X laevis. Serial adherent subculture resulted in spontaneously immortalized cells and the establishment of two MG cell lines: murine retinal glia 17 (RG17) and Xenopus glia 69 (XG69). They were characterized for MG gene and protein expression by qPCR, immunostaining, and Western blot. Purinergic signaling was assessed with calcium imaging. Pharmacological perturbations with 2'-3'-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) and KN-62 were performed on RG17 cells. Results: RG17 and XG69 cells express several MG markers and retain purinergic signaling. Pharmacological perturbations of intracellular calcium responses with BzATP and KN-62 suggest that the ionotropic purinergic receptor P2X7 is present and functional in RG17 cells. Stimulation of XG69 cells with adenosine triphosphate-induced calcium responses in a dose-dependent manner. Conclusions: We report the characterization of RG17 and XG69, two novel MG cell lines from species with significantly disparate retinal regenerative capabilities. Translational Relevance: RG17 and XG69 cell line models will aid comparative studies between species endowed with varied regenerative capacity and will facilitate the development of new cell-based strategies for treating retinal degenerative diseases.

The Potential Cutaneous Effects of Pornography Addiction: A Narrative Review.

Pornography addiction is an area of increasing concern, particularly due to the ubiquitous nature of pornographic material on the Internet. Even so, there is no formal Diagnostic and Statistical Manual of Mental Disorders (DSM-5) inclusion of compulsive pornography use as a behavioral addiction. Although the psychosocial impacts of pornography addiction have been studied, the risk of direct skin injuries and behavioral changes brought about by excessive pornography usage remain to be seen. Adult males constitute the majority of cases of engaging in risky and violent sexual behaviors with an unclear association with pornography consumption. Adult females may be likely to copy pubic hair trimming patterns analogous to those seen in pornographic content, even though attitudes towards labiaplasty are unclear. Finally, adolescents regularly exposed to pornography have been found to replicate sexual activity seen in pornographic material and have earlier sexual activity. In the literature evaluated, an association between pornographic material and direct cutaneous disease remains a major area of further research.

Association amid the comorbidity of Diabetes Mellitus in patients of Active Tuberculosis in Pakistan: A matched case control study.

OBJECTIVES: This study aims to test the association between diabetes and tuberculosis. METHODS: It is a matched case control study conducted in tertiary care hospitals in 2019-2020. Cases and controls were 144 each, selected on the basis of an odds ratio of 2 at 95% confidence interval with a significance level of 5%. Cases of pulmonary tuberculosis were selected through consecutive sampling technique, either visiting OPD or admitted in hospital. Controls were taken from the general population and frequency matching was done based on age, gender and socioeconomic status. Data was collected through structured questionnaire after taking written consent. Data was analyzed on SPSS version 23. Binary Logistic regression model was applied for finding association between the risk factors and the disease. P value <0.05 was considered statistically significant. RESULTS: Out of all cases and controls, 45% and 20% were diabetics respectively. The association between the risk factors and tuberculosis was estimated by univariate analysis, positive association was found between diabetes and tuberculosis (OR= 3.32), a high frequency of diabetes in cases as compared to controls were observed with a highly significant p- value (<.001). CONCLUSIONS: This study provides evidence for a strong positive association between tuberculosis and diabetes.

Association of subjective memory complaints amid patients of Diabetes Mellitus Type II and Hypertension.

BACKGROUND AND OBJECTIVE: Subjective memory complaints (SMCs) are suggested to predict dementia at a very early stage. However, association of SMCs with known risk factors of dementia namely diabetes mellitus Type-2 diabetes and hypertension (HTN) remain unexplored which is the main aim of this study. The objective of the study was to investigate the association of SMC with diabetes mellitus type 2 (DM2) and hypertension (HTN). METHODS: The associations of diabetes and hypertension, with SMCs has been tested in 500 participants from a tertiary care hospital of Karachi during 2017. Diagnosed cases of diabetes and hypertension were included through convenient sampling. Healthy attendants of patients were interviewed for the reference group. SMCs were assessed through a 14-item SMC questionnaire. RESULTS: Sample included 114 patients with only diabetes and hypertension each, 103 with both diabetes and hypertension and 169 healthy participants. Compared to healthy adults, persons with diabetes and hypertension had higher SMCs (difference, 0.88, 95% CI: 0.22, 1.54) (difference, 1.06, 95% CI: 0.40, 1.71) respectively, in fully adjusted models. CONCLUSIONS: Compared to healthy adults of working age-group, persons with diabetes and hypertension were more likely to have SMCs. Assessment and early detection of SMCs in persons with diabetes and hypertension might be informative to provide a window for effective interventions to maintain cognitive health.

Secretory Functions of Macrophages in the Human Pancreatic Islet Are Regulated by Endogenous Purinergic Signaling.

Endocrine cells of the pancreatic islet interact with their microenvironment to maintain tissue homeostasis. Communication with local macrophages is particularly important in this context, but the homeostatic functions of human islet macrophages are not known. In this study, we show that the human islet contains macrophages in perivascular regions that are the main local source of the anti-inflammatory cytokine interleukin-10 (IL-10) and the metalloproteinase MMP9. Macrophage production and secretion of these homeostatic factors are controlled by endogenous purinergic signals. In obese and diabetic states, macrophage expression of purinergic receptors MMP9 and IL-10 is reduced. We propose that in those states, exacerbated ß-cell activity due to increased insulin demand and increased cell death produce high levels of ATP that downregulate purinergic receptor expression. Loss of ATP sensing in macrophages may reduce their secretory capacity.

MRI and histological evaluation of pulsed focused ultrasound and microbubbles treatment effects in the brain.

Magnetic resonance imaging (MRI)-guided pulsed focused ultrasound (pFUS) combined with microbubbles (MB) contrast agent infusion has been shown to transiently disrupt the blood-brain barrier (BBBD), increasing the delivery of neurotherapeutics to treat central nervous system (CNS) diseases. pFUS interaction with the intravascular MB results in acoustic cavitation forces passing through the neurovascular unit (NVU), inducing BBBD detected on contrast-enhanced MRI. Multiple pFUS+MB exposures in Alzheimer's disease (AD) models are being investigated as a method to clear amyloid plaques by activated microglia or infiltrating immune cells. Since it has been reported that pFUS+MB can induce a sterile inflammatory response (SIR) [1-5] in the rat, the goal of this study was to investigate the potential long-term effects of SIR in the brain following single and six weekly sonications by serial high-resolution MRI and pathology. Methods: Female Sprague Dawley rats weighing 217±16.6 g prior to sonication received bromo-deoxyuridine (BrdU) to tag proliferating cells in the brain. pFUS was performed at 548 kHz, ultrasound burst 10 ms and initial peak negative pressure of 0.3 MPa (in water) for 120 s coupled with a slow infusion of ~460 µL/kg (5-8×107 MB) that started 30 s before and 30 s during sonication. Nine 2 mm focal regions in the left cortex and four regions over the right hippocampus were treated with pFUS+MB. Serial high-resolution brain MRIs at 3 T and 9.4 T were obtained following a single or during the course of six weekly pFUS+MB resulting in BBBD in the left cortex and the right hippocampus. Animals were monitored over 7 to 13 weeks and imaging results were compared to histology. Results: Fewer than half of the rats receiving a single pFUS+MB exposure displayed hypointense voxels on T2*-weighted (w) MRI at week 7 or 13 in the cortex or hippocampus without differences compared to the contralateral side on histograms of T2* maps. Single sonicated rats had evidence of limited microglia activation on pathology compared to the contralateral hemisphere. Six weekly pFUS+MB treatments resulted in pathological changes on T2*w images with multiple hypointense regions, cortical atrophy, along with 50% of rats having persistent BBBD and astrogliosis by MRI. Pathologic analysis of the multiple sonicated animals demonstrated the presence of metallophagocytic Prussian blue-positive cells in the parenchyma with significantly (p<0.05) increased areas of activated astrocytes and microglia, and high numbers of systemic infiltrating CD68+ macrophages along with BrdU+ cells compared to contralateral brain. In addition, multiple treatments caused an increase in the number of hyperphosphorylated Tau (pTau)-positive neurons containing neurofibrillary tangles (NFT) in the sonicated cortex but not in the hippocampus when compared to contralateral brain, which was confirmed by Western blot (WB) (p<0.04). Conclusions: The repeated SIR following multiple pFUS+MB treatments could contribute to changes on MR imaging including persistent BBBD, cortical atrophy, and hypointense voxels on T2w and T2*w images consistent with pathological injury. Moreover, areas of astrogliosis, activated microglia, along with higher numbers of CD68+ infiltrating macrophages and BrdU+ cells were detected in multiple sonicated areas of the cortex and hippocampus. Elevations in pTau and NFT were detected in neurons of the multiple sonicated cortex. Minimal changes on MRI and histology were observed in single pFUS+MB-treated rats at 7 and 13 weeks post sonication. In comparison, animals that received 6 weekly sonications demonstrated evidence on MRI and histology of vascular damage, inflammation and neurodegeneration associated with the NVU commonly observed in trauma. Further investigation is recommended of the long-term effects of multiple pFUS+MB in clinical trials.

First aid facilities in the school settings: Are schools able to manage adequately?

BACKGROUND AND OBJECTIVE: Children spend most of their time in schools and are vulnerable to injuries and mild ailments, hence requiring first-aid care. School teacher can provide immediate first-aid care in the absence of any health professional. This study assesses first-aid facilities within school premises and assessment of teachers on first aid training. METHODS: A cross sectional study was conducted from July-December 2017, participants were full time school teachers of both public and private sectors at both primary and secondary levels, having a minimum of one year experience. Questionnaire was filled on one to one basis by taking oral interview. RESULTS: Out of 209 teachers, 72.7% were from private sector. Stomachache was the most common medical incident (82.29%) requiring first-aid care in schools. First aid box was available in all schools but its contents were not satisfactory. Sick bay was not found in any school. 68.42% of teachers were not trained in first-aid management because of lack of opportunity, however 56% were willing to enroll in any first aid training and majority (91.38%) considered it essential for their professional life. CONCLUSION: First aid facilities at various schools of Karachi and availability of trained teachers who can provide first aid care is unsatisfactory.

Disrupting the blood-brain barrier by focused ultrasound induces sterile inflammation.

MRI-guided pulsed focused ultrasound (pFUS) combined with systemic infusion of ultrasound contrast agent microbubbles (MB) causes localized blood-brain barrier (BBB) disruption that is currently being advocated for increasing drug or gene delivery in neurological diseases. The mechanical acoustic cavitation effects of opening the BBB by low-intensity pFUS+MB, as evidenced by contrast-enhanced MRI, resulted in an immediate damage-associated molecular pattern (DAMP) response including elevations in heat-shock protein 70, IL-1, IL-18, and TNFα indicative of a sterile inflammatory response (SIR) in the parenchyma. Concurrent with DAMP presentation, significant elevations in proinflammatory, antiinflammatory, and trophic factors along with neurotrophic and neurogenesis factors were detected; these elevations lasted 24 h. Transcriptomic analysis of sonicated brain supported the proteomic findings and indicated that the SIR was facilitated through the induction of the NFκB pathway. Histological evaluation demonstrated increased albumin in the parenchyma that cleared by 24 h along with TUNEL+ neurons, activated astrocytes, microglia, and increased cell adhesion molecules in the vasculature. Infusion of fluorescent beads 3 d before pFUS+MB revealed the infiltration of CD68+ macrophages at 6 d postsonication, as is consistent with an innate immune response. pFUS+MB is being considered as part of a noninvasive adjuvant treatment for malignancy or neurodegenerative diseases. These results demonstrate that pFUS+MB induces an SIR compatible with ischemia or mild traumatic brain injury. Further investigation will be required before this approach can be widely implemented in clinical trials.

Iatrogenic foreign body in the urinary bladder.

A 60 years old man presented with a history of suprapubic pain, painful micturition, perineal pain, and acute retention of urine followed by catheterization. On abdominal ultrasound, there was a metallic needle found in the urinary bladder which was confirmed on pelvis X-ray. The needle was found to be a metallic piece of a resectoscope cutting loop which was removed through cystourethroscopy.

Stress-induced dissociations between intracellular calcium signaling and insulin secretion in pancreatic islets.

In healthy pancreatic islets, glucose-stimulated changes in intracellular calcium ([Ca(2+)]i) provide a reasonable reflection of the patterns and relative amounts of insulin secretion. We report that [Ca(2+)]i in islets under stress, however, dissociates with insulin release in different ways for different stressors. Islets were exposed for 48h to a variety of stressors: cytokines (low-grade inflammation), 28mM glucose (28G, glucotoxicity), free fatty acids (FFAs, lipotoxicity), thapsigargin (ER stress), or rotenone (mitochondrial stress). We then measured [Ca(2+)]i and insulin release in parallel studies. Islets exposed to all stressors except rotenone displayed significantly elevated [Ca(2+)]i in low glucose, however, increased insulin secretion was only observed for 28G due to increased nifedipine-sensitive calcium-channel flux. Following 3-11mM glucose stimulation, all stressors substantially reduced the peak glucose-stimulated [Ca(2+)]i response (first phase). Thapsigargin and cytokines also substantially impacted aspects of calcium influx and ER calcium handling. Stressors did not significantly impact insulin secretion in 11mM glucose for any stressor, although FFAs showed a borderline reduction, which contributed to a significant decrease in the stimulation index (11:3mM glucose) observed for FFAs and also for 28G. We also clamped [Ca(2+)]i using 30mM KCl+250µM diazoxide to test the amplifying pathway. Only rotenone-treated islets showed a robust increase in 3-11mM glucose-stimulated insulin secretion under clamped conditions, suggesting that low-level mitochondrial stress might activate the metabolic amplifying pathway. We conclude that different stressors dissociate [Ca(2+)]i from insulin secretion differently: ER stressors (thapsigargin, cytokines) primarily affect [Ca(2+)]i but not conventional insulin secretion and 'metabolic' stressors (FFAs, 28G, rotenone) impacted insulin secretion.

Periocular Scleromyxedema.

Scleromyxedema is characterized by cutaneous mucinosis and rarely presents to oculoplastic surgeons with bilateral upper and lower eyelid swelling. The authors present 2 case reports with a review of the literature and discuss the ophthalmic manifestations of scleromyxedema and the management of these cases. Both patients were in their early 50s and were fit and well prior to the presentation. They both presented with eyelid swelling and underlying nodularity of the subcutaneous tissue. The skin biopsy confirmed cutaneous manifestations of scleromyxedema. The systemic manifestations responded to intravenous and oral steroids and intravenous infusions of immunoglobulin. The ophthalmic manifestations were managed conservatively. Scleromyxedema is a rare connective tissue disease characterized by cutaneous mucinosis, extracutaneous manifestations, and monoclonal gammopathy. It rarely affects the eyelids and cornea. The authors would like to present 2 cases of periocular scleromyxedema.

Reflux of drug during intra-vitreal anti-VEGF therapies.

PURPOSE: To report reflux of anti-VEGF drug during intravitreal injections. METHOD: Review of electronic case notes of patients undergoing intra-vitreal anti-VEGF treatments. Prospective data collection was performed with a specific emphasis on presence or absence of reflux. RESULTS: 152 records of 102 eyes were available from an 8-month period. 119 Ranibizumab injections and 33 bevazicumab injections were considered. Reflux was noted in 48 injections (31%). For eyes with first injection of anti-VEGF agent, reflux was noted in 9/23 eyes. Eyes with at least one previous injection were observed to have reflux in 24/79 eyes. Presence of reflux was statistically tested against posterior vitreous detachment and phakic status and was found to be statistically insignificant. CONCLUSION: Intra-vitreal anti-VEGF injections with volumes of 0.05 ml appears to produce displacement of the conjunctiva with a transient fluid-filled bleb immediately after the injection in approximately 1/3 of eyes.

Macular infarction after inadvertent intracameral cefuroxime.

UNLABELLED: We present the case of a 70-year-old patient who had uneventful cataract surgery. Because of a procedural mishap, the patient received an excess dose of intracameral cefuroxime of approximately 62.5 mg. Two weeks postoperatively, evidence of macular infarction with cystoid macular edema was seen on fundus fluorescein angiography. The patient was treated with 4.0 mg of intravitreal triamcinolone, but the visual acuity failed to improve; the final pinhole acuity was 3/60. Based on the postoperative progress, we speculate that a delayed mechanism of vascular toxicity is implicated. The importance of systematic procedures to reduce the risk for mistakes is emphasized, especially for a procedure such as cataract surgery where efficiency is increasingly important. FINANCIAL DISCLOSURE: Neither author has a financial or proprietary interest in any material or method mentioned.

Congenital isolated absence of the inferior oblique muscle and compensatory aberrant insertion of the inferior and lateral recti.

Absence of various extra-ocular muscles has previously been described. However isolated absence of the inferior oblique has not yet been described. Our patient was found to have a small right esotropia and limitation of right eye abduction with an abnormal head posture. Ocular movements showed a marked adduction overshoot in elevation. Exploration of the musculature revealed an absent inferior oblique with abnormally inserted and tight inferior and lateral rectus muscles. Imaging, forced duction testing and surgical exploration is recomended in cases where the signs do not fit into a clear syndrome.

Primary intravitreal ranibizumab for myopic choroidial neovascularisation.

Myopic chorodial neovascularisation (mCNV) is an important cause of visual loss in high myopia with almost 10% of eyes with pathological myopia developing mCNV. Intravitreal anti-VEGF agents have recently been documented to be effective in mCNV. However, controversy exists regarding the optimal drug, its dose, and the frequency of administration. We performed a retrospective case review examining patients who had myopia of less than -6D and a chorodial neovascular membrane diagnosed on fundus fluorescein angiography (FFA) that were treated with 3 injections of ranibizumab (Lucentis) 5mg/0.05ml given at monthly intervals. The course was repeated if the ocular coherence tomography (OCT) continued to show intra-retinal oedema at review. We recorded logMAR visual acuities and central foveal thickness (CFT) at baseline and final review. Improvement in visual acuity was significantly improved (p = 0.049) by a mean of 0.24 logMAR (range 0 to 0.74). Every patient achieved at least visual stability. Mean CFT reduction was also significantly improved (p = 0.02) by a mean of 109 microns (range -8 to 198). Our series and current literature seem to support as primary treatment a standard dose of an anti-VEGF agent. Further, larger studies are required to clarify whether any particular injection strategy is clearly superior.

Effect of reflux of drug during intravitreal anti-VEGF therapies on foveal thickness.

PURPOSE: To report any variation in foveal thickness in eyes with and without reflux during anti-VEGF injection. METHOD: Review of electronic case notes and scans. RESULTS: 18 patients receiving bevazicumab were identified as having reflux (n = 3) or no reflux (n = 15). Pre-injection average central foveal thickness (CFT) was 439 microns (µ) and post-injection average CFT was 417µ. The change in mean CFT was -22 µ (range -330 to 336 µ). 70 patients receiving ranibizumab (lucentis) were identified as having reflux (n = 22) or no reflux (n = 48). Average pre-injection CFT was 336 µ and post-injection average CFT was 289 µ. The change in mean CFT was -48 µ (range -163 to 443 µ). The change in CFT between the group with reflux and no reflux in bevazicumab and ranibizumab injections was statistically insignificant. CONCLUSION: Reflux following injection of anti-VEGF agents does not appear to cause a sub-therapeutic effect.