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Abstract Functional genomics, a multidisciplinary subject, investigates the functions of genes and their products in biological systems to better understand diseases and find new drugs. Drug repurposing is an economically efficient approach that entails discovering novel therapeutic applications for already-available medications. Genomics enables the identification of illness and therapeutic molecular characteristics and interactions, which in turn facilitates the process of drug repurposing. Techniques like gene expression profiling and Mendelian randomization are helpful in identifying possible medication candidates. Progress in computer science allows for the investigation and modeling of gene expression networks that involve large amounts of data. The amalgamation of data concerning DNA, RNA, and protein functions bears similarity to pharmacogenomics, a crucial aspect in crafting cancer therapeutics. Functional genomics in drug discovery, particularly for cancer, is still not thoroughly investigated, despite the existence of a significant amount of literature on the subject. Next-generation sequencing and proteomics present highly intriguing opportunities. Publicly available databases and mining techniques facilitate the development of cancer treatments based on functional genomics. Broadening the exploration and utilization of functional genomics holds significant potential for advancing drug discovery and repurposing, particularly within the realm of oncology.

Worldwide, the harmful ion contamination of water has become a serious problem because of unregulated industry, energy production, and mining, which greatly increase the concentration of pollutants in water. The novel membranes through adsorbent self-assembly, such as protein amyloids, were explored for wastewater treatment. Herein, we report amyloid fibril (AF)-embedded bacterial cellulose nanohybrid membrane for efficient removal of heavy metal from industrial effluent. AFs are synthesized by heat treatment using bovine serum albumin and embedded with bacterial cellulose nanomembrane (BCN). The AF-embedded BCN (AF/BCN) was characterized using microscopy and spectroscopic methods. In addition, the well-ordered multi-layered AF/BCN filtration assembly was fabricated in the commercial cartridge and validated for the removal of heavy metals (Pb2+ and Hg2+) from wastewater and treatment of industrial wastewater sample containing heavy metals. Our multi-layered filtration assembly removed Hg2+ and Pb2+ with efficiency of 95 and 78.34%, respectively. A computational study using molecular docking has also been performed for the identification of metal entrapment sites. Moreover, our AF/BCN filtration assembly showed high regeneration capacity up to four cycles. The isotherm model also revealed a strong fit and good adsorption behaviour. This makes potential filtration assembly for low-cost, high-efficiency for the removal of heavy metal from wastewater. HIGHLIGHTS Computational approaches methods for the study of bovine serum albumin.; Synthesis of amyloid fibril, amyloid-embedded bacterial cellulose nanomembrane (BCN), and fabrication of amyloid-embedded BCN filtration assembly.; Heavy metal removal analysis from industrial wastewater.;

Abstract Cancer has been viewed as one of the deadliest diseases worldwide. Among various types of cancer, breast cancer is the most common type of cancer in women. Methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is a promising druggable target and is overexpressed in cancerous cells, like, breast cancer. We conducted structure-based modeling on the allosteric site of the enzyme. Targeting the allosteric site avoids the problem of drug resistance. Pharmacophore modeling, molecular docking, HYDE assessment, drug-likeness, ADMET predictions, simulations, and free-energy calculations were performed. The RMSD, RMSF, RoG, SASA, and Hydrogen-bonding studies showed that seven candidates displayed stable behaviour. As per the literature, average superimposed simulated structures revealed a similar protein conformational change in the αEʹ-βfʹ loop, causing its displacement away from the allosteric site. The MM-PBSA showed tight binding of six compounds with the allosteric pocket. The effect of inhibitors interacting in the allosteric site causes a decrease in the binding energy of J49 (active-site inhibitor), suggesting the effect of allosteric binding. The PCA and FEL analysis revealed the significance of the docked compounds in the stable behaviour of the complexes. The outcome can contribute to the development of potential natural products with drug-like properties that can inhibit the MTHFD2 enzyme.

Abstract Background The physical challenges faced by adolescents and young adults (AYA) after a cancer diagnosis may be different from those experienced by paediatric and older adult cancer patients. Patient‐reported outcome measures (PROMs) are valuable tools that can be useful in exploring the experiences of AYAs and identifying important issues, recurrent themes and areas to potentially improve quality of life. Objective We compared patient‐reported physical function outcomes between AYAs diagnosed with cancer and non‐cancer controls. Method This paper builds on a scoping review published in early 2023 and focuses on PROMs related to physical function. Results This systematic review includes 16 studies that measured and reported on physical function PROMs in AYA cancer survivors compared with their cancer‐free peers. Of these studies, 14 found that physical function in AYA survivors was significantly worse. This paper also includes a meta‐analysis conducted on 5 studies using the EORTC‐QLQ‐C30 to measure physical function, which found that physical function score was an average of 7.03 (95% CI: −10.21, −3.86) points lower in the AYA cancer group, compared to their cancer free‐peers, a difference that is clinically meaningful. Conclusions The results overwhelmingly demonstrate that AYAs post a cancer diagnosis have worse health‐related quality of life from a physical function perspective than their cancer‐free peers, providing a compelling argument for the need to address this issue. All but one of the studies were cross‐sectional, which highlights the need for further assessment of this group longitudinally throughout their cancer journey.

Introduction: The most important acquired brain injuries in very and extremely preterm infants born in developing Country are periventricular-Intraventricular haemorrhages and diffuse white matter injury. Antenatal corticosteroids play a crucial role in its development. A study was conducted to determine effects of antenatal steroids along with magnesium sulphate on Intraventricular Hemorrhage and Periventricular Leukomalacia in preterm neonates. Methodology: A retrospective prospective cohort study conducted among premature newborn babies admitted in the hospital. Based on the antenatal medication subjects were divided in to three groups: Group 1: No Steroid and No MgsO4 in antenatal period; Group 2: Received complete and incomplete course of antenatal steroid; and Group 3: Received complete course of antenatal steroid and MgSO4. The Occurrence of IVH and/or PVL in the neonates during hospital stay in all 3 groups. Results: Among the 144 cases, 8 patients had IVH and 1 had PVL. No intervention group (No antenatal steroids/ antenatal MgSo4) had highest chances of development of IVH/PVL. Significantly lower rate of IVH/PVL recorded in ‘Steroid and MgSO4’ group compared to only ‘Steroid’ group. The chances of development of RDS, BPD and NEC in no intervention group (No antenatal steroids and antenatal MgSo4) was significantly higher compared to steroid and and MgSO4 group and Steroid only group. Conclusion: This study confirmed that antenatal steroids when used along with antenatal magnesium sulphate to deliver preterm babies reduce IVH and white matter brain injury significantly.

Wnt signaling is a critical pathway involved in cell proliferation, differentiation and cellular homeostasis. To address treatment need of diseases associated with the dysregulated Wnt signaling like cancer, Alzheimer’s disease, Osteoporosis, Myocardial infarction etc.; small molecules that target the very first and unique component of the Wnt pathway, Porcupine enzyme, have been proven to be effective. Till date, none of the Porcupine inhibitor has reached to market. To design the novel Porcupine inhibitors, there was a need of crystal structure of Porcupine but in 2019, it was not resolved and deposited in the protein data bank. So, with an aim to predict its 3D structure, homology modeling study was performed using two distinct platforms; I-TASSER and Molsoft ICMPro so that we could use the best validated model for the designing of NCEs. Both the generated homology models were compared through Ramachandran plot, Protein health tool of Molsoft ICM and other tools available on metaserver, SAVES v6.0. Molsoft model was found better with 84.6 % residues in most favored region and only 0.3 % residues in disallowed region in Ramachandran plot in comparison to 75.9 % and 1.7 % residues respectively in I-TASSER model. The ICM model was further refined for 50 ns under MD simulation where it got stabilized after 36 ns. Quantification of protein was done by Radius of Gyration (ROG) where model showed larger ROG value of 23.25°A which indicated closely packed structure. Binding site in the predicted structure of Porcupine was identified using ICM binding pocket identifier and used for molecular docking of known Porcupine inhibitors like IWP-2, IWP-3, IWP-L6, and LKG974. Key binding site residues; Arg124, Phe257, Leu261, Val302, Trp305, Asn306, Ser310, Leu313, His341, Phen345, Ala349, Val350 etc. involved in the interaction were identified after MD simulation which can be useful in designing of novel porcupine inhibitors in future.

Drug administration by oral delivery is the preferred route, regardless of some remaining challenges, such as short resident time and toxicity issues. One strategy to overcome these barriers is utilizing mucoadhesive vectors that can increase intestinal resident time and systemic uptake. In this study, biomimetic nanoparticles (NPs) were produced from 14 types of edible algae and evaluated for usage as oral DDSs by measuring their size, surface charge, morphology, encapsulation efficiency, mucoadhesion force, and cellular uptake into Caco-2 cells. The NPs composed of algal materials (aNPs) exhibited a spherical morphology with a size range of 126–606 nm and a surface charge of −9 to −38 mV. The mucoadhesive forces tested ex vivo against mice, pigs, and sheep intestines revealed significant variation between algae and animal models. Notably, Arthospira platensis (i.e., Spirulina) NPs (126 ± 2 nm, −38 ± 3 mV) consistently exhibited the highest mucoadhesive forces (up to 3127 ± 272 µN/mm²). Moreover, a correlation was found between high mucoadhesive force and high cellular uptake into Caco-2 cells, further supporting the potential of aNPs by indicating their ability to facilitate drug absorption into the human intestinal epithelium. The results presented herein serve as a proof of concept for the possibility of aNPs as oral drug delivery vehicles.

Human immunodeficiency virus (HIV)-exposed infants may be at increased risk of vaccine-preventable disease. This study was conducted as a post-licensure commitment in this population to evaluate the primary series, antibody persistence, and booster response to a licensed fully liquid hexavalent vaccine containing diphtheria (D), tetanus (T), acellular pertussis (aP), inactivated poliovirus (IPV), hepatitis B (HB), and Haemophilus influenzae type b antigens (PRP~T). This was a Phase III, open-label, randomized study conducted at a single center in the Republic of South Africa. The DTaP-IPV-HB-PRP~T vaccine was administered to HIV-exposed infected (Group A: N = 14) and HIV-exposed uninfected (Group B: N = 50) infants as a 6, 10, 14 week primary series with a toddler booster at 15–18 months of age. Immunogenicity of each antigen was measured using validated assays and vaccine reactogenicity was recorded using diary cards. The low number of HIV-exposed infected participants, due to widespread pre- and peri-natal retroviral treatment, meant that between-group comparisons should be treated with caution. In each group, primary series and booster immune seroprotection rates were strong, and pre-booster antibody persistence was good, although anti-HBs ≥10 mIU/mL in Group A was 78.6% post-primary series, 58.3% pre-booster, and 75.0% post-booster. There were no safety concerns. In conclusion, primary series and booster vaccination of the DTaP-IPV-HB-PRP~T vaccine were immunogenic and safe in HIV-exposed infected and uninfected infants. These results were comparable to historical data in healthy infants and toddlers.

Abstract The protein alpha-synuclein (α-syn) is unusual because, depending on its conformation and the type of cell in which it is expressed, it is pro-death or pro-survival, triggering neurodegeneration in Parkinson’s disease and enhancing cell survival of some melanomas. To probe the function of α-syn in melanoma, we used CRISPR/Cas9 to knockout SNCA, the gene that codes for α-syn, in SK-Mel-28 melanoma cells. The SNCA-knockout clones in culture exhibited a decrease in the transferrin receptor 1 (TfR1), an increase in ferritin, an increase of reactive oxygen species and proliferated slower than control cells. These SNCA-knockout clones grafted into SCID mice grew significantly slower than the SK-Mel-28 control cells that expressed α-syn. In the excised SNCA-knockout xenografts, TfR1 decreased 3.3-fold, ferritin increased 6.2-fold, the divalent metal ion transporter 1 (DMT1) increased threefold, and the iron exporter ferroportin (FPN1) decreased twofold relative to control xenografts. The excised SNCA-KO tumors exhibited significantly more ferric iron and TUNEL staining relative to the control melanoma xenografts. Collectively, depletion of α-syn in SK-Mel-28 cells dysregulates cellular iron metabolism, especially in xenografts, yielding melanoma cells that are deficient in TfR1 and FPN1, that accumulate ferric iron and ferritin, and that undergo apoptosis relative to control cells expressing α-syn.

Context: Snakebite remains an underrated cause of accidental death in modern India, primarily in rural India, where people fail to reach out to modern medicine and fall victim to the handful of quacks using traditional healing methods. If promptly diagnosed and treated based on various clinical determinants like mode of presentation, time of medical intervention, recognition of the species, and analysis of a series of reliably identified bites, the treatment outcome would be more promising. We aimed to study snakebite patients' clinical profile and treatment outcome in a rural tertiary care setup. Materials and Method: This is a retrospective study in which the data evaluated from an epidemiological viewpoint; gender and age of the snake bite victim, time when bitten, interval between the bite and medical consultation, pattern of toxicity, and response to anti-snake venom (ASV). Results: Of a total of 200 patients bitten by a snake, 121 were males, with 77% adults. In nearly all cases, the type of snake was unknown; however, most of the bites were poisonous, showing one or the other type of toxicity. One hundred seventy-one patients survived the snake bite, and 29 succumbed. When Logistic regression was done with Death/discharge as the dependent variable and “Time to bite and reaching hospital, Age, Sex, number of ASV given, Ventilation needed or not, pack cell volume (PCV) numbers, Fresh Frozen Plasma (FFP) numbers, Dialysis and presence or absence of toxicity” as the independent variables, the model developed did not account for any respectable amount of variation in the outcome. The only variable found to be predicting the outcome significantly was FFP. Conclusion: It is often difficult to identify the type of snake, and thus polyvalent antisnake venom remains the only available treatment resource. Readily available treatment resources, timely intervention, appropriate referral, and close ICU will alleviate mortality.

Objectives The impending and increasing prevalence of diabetic retinopathy (DR) in India has necessitated a need for affordable and valid community outreach screening programme for DR, especially in rural and far to reach indigenous local communities. The present study is a pilot study aimed to compare non-mydriatic fundus photography with indirect ophthalmoscopy for its utilisation as a feasible and logistically convenient screening modality for DR in an older age, rural, tribal population in Western India.Design and setting This community-based, cross-sectional, prospective population study was a part of a module using Rapid Assessment of Avoidable Blindness and DR methodology in 8340 sampled participants with ≥50 years age. In this study, the diabetics identified were screened for DR using two methods: non-mydriatic fundus photography on the field by trained professionals, that were then graded by a retina specialist at the base hospital and indirect ophthalmoscopy by expert ophthalmologists in the field with masking of each other’s findings for its utility and comparison.Results The prevalence of DR, sight threatening DR and maculopathy using indirect ophthalmoscopy was found to be 12.1%, 2.1% and 6.6%, respectively. A fair agreement (κ=0.48 for DR and 0.59 for maculopathy) was observed between both the detection methods. The sensitivity and specificity of fundus photographic evaluation compared with indirect ophthalmoscopy were found to be 54.8% and 92.1% (for DR), 60.7% and 90.8% (for any DR) and 84.2% and 94.8% (for only maculopathy), respectively.Conclusion Non-mydriatic fundus photography has the potential to identify DR (any retinopathy or maculopathy) in community settings in Indian population. Its utility as an affordable and logistically convenient cum practical modality is demonstrable. The sensitivity of this screening modality can be further increased by investing in better resolution cameras, capturing quality images and training and validation of imagers.Trial registration number CTRI/2020/01/023025; Clinical Trial Registry, India (CTRI).

Abstract Background Hirudo granulosa—an Indian cattle leech, is frequently used for the treatment of psoriasis and eczema. During treatment which followed by leech biting, it transfers some amount of gut microbes along with saliva. This may result in bacterial infection at the treatment site. Antibiotics used as post-surgical hirudotherapy is the reason for drug resistance. Drug resistance is the result of the change in the genetic makeup of bacteria. Therefore, it is necessary to study antibiotic sensitivity of gut bacteria and characterized them genetically. Results Fourteen bacterial isolates were obtained from unused leech in which five were Gram-negative and the other nine were Gram-positive. Similarly from the used leech, thirteen were isolated in which five were Gram-negative and the other eight were Gram-positive. Biochemical analysis reveals that isolates from unused leech saliva belong to Pseudomonas, Micrococcus, Streptococcus, and Vibrio species, while the used leech salivary bacteria were the member of genus Pseudomonas, Comamonas, Escherichia, Citrobacter, Aeromonas, Providencia, Enterobacter, and Yersinia. Antibiotic sensitivity tests for isolates indicated that chloramphenicol (30 μg) and norfloxacin (10 μg) were effective for unused leech isolates while sparfloxacin (5 μg) and Cefaclor (30 μg) were effective against used leech salivary isolates. Random primer (OPL-14)-based random amplified polymorphic DNA (RAPD) fingerprint showed twenty amplified regions among all tested bacteria. Most of the bacteria contain the tested sequence except U2, T2, T9A, and T10 which showed no amplification indicated the absence of primers sequence. Conclusion Chloramphenicol, norfloxacin, sparfloxacin, and Cefaclor antibiotics alone or in combination were possibly used to treat post-therapy infections. Bacteria from treated and untreated leeches were clustered at nearby branch in neighbor end-joining phylogenetic tree, which indicates the similar (but not exact) genetic makeup. Therefore, it can be concluded that these antibiotics were possibly used against most of them.

Pheochromocytoma (PHEO) in multiple endocrine neoplasia type 1 (MEN1) is extremely rare. The incidence is reported as less than 2%. We report a case of a 76-year-old male with familial MEN1 who was found to have unilateral PHEO. Although the patient was normotensive and asymptomatic, routine screening imaging with CT demonstrated bilateral adrenal masses. The left adrenal mass grew from 2.5 to 3.9 cm over 4 years with attenuation values of 9 Hounsfield units (HU) pre-contrast and 15 HU post-contrast washout. Laboratory evaluation demonstrated an adrenergic biochemical phenotype. Both 18F-fluorodeoxyglucose (18F-FDG) PET/CT and 123I-metaiodobenzylguanidine (123I-mIBG) scintigraphy demonstrated bilateral adrenal uptake. In contrast, 18F-fluorodihydroxyphenylalanine (18F-FDOPA) PET/CT demonstrated unilateral left adrenal uptake (28.7 standardized uptake value (SUV)) and physiologic right adrenal uptake. The patient underwent an uneventful left adrenalectomy with pathology consistent for PHEO. Post-operatively, he had biochemical normalization. A review of the literature suggests that adrenal tumors >2 cm may be at higher risk for pheochromocytoma in patients with MEN1. Despite a lack of symptoms related to catecholamine excess, enlarging adrenal nodules should be biochemically screened for PHEO. 18F-FDOPA PET/CT may be beneficial for localization in these patients.

Eales' disease (ED), which is an idiopathic obliterative vasculitis, is a diagnosis of exclusion. The optic nerve involvement in ED is not very common. We report a case of ED in a 36-year-old male who initially presented as papillephlebitis. He presented with complaints of decreased vision in his right eye for 2 months. Fundus examination revealed that optic disc edema with hemorrhages, and he was started on oral corticosteroid after the exclusion of infectious etiology. Fundus examination after 2 months revealed thet resolution of optic disc edema, but active periphlebitis with multiple superficial retinal hemorrhages involving inferior and inferotemporal quadrant. Based on negative laboratory results and clinical findings a diagnosis of ED was considered. Regular monitoring of patients with papillephlebitis is recommended.

Novel severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) has claimed more than 3.3 million lives worldwide and still counting. As per the GISAID database, the genomics of SARS-CoV-2 has been extensively studied, with more than 500 genome submissions per day. Out of several hotspot mutations within the SARS-CoV-2 genome, recent research has focused mainly on the missense variants. Moreover, significantly less attention has been accorded to delineate the role of the untranslated regions (UTRs) of the SARS-CoV-2 genome in the disease progression and etiology. One of the most frequent 5’ UTR variants in the SARS-CoV-2 genome is the C241T, with a global frequency of more than 95 %. In the present study, the effect of the C241T mutation has been studied with respect to the changes in RNA structure and its interaction with the host replication factors MADP1 Zinc finger CCHC-type and RNA-binding motif 1 (hnRNP1). The results obtained from molecular docking and molecular dynamics simulation indicated weaker interaction of C241T mutant stem-loops with the host transcription factor MADP1, indicating a reduced replication efficiency. The results are also correlated with increased recovery rates and decreased death rates of global SARS-CoV-2 cases.

Sickle cell disease is a progressively debilitating genetic condition that affects red blood cells and can result in a variety of serious medical complications, reduced life expectancy, and diminished quality of life. Medicaid nationwide covered 66 percent of sickle cell disease hospitalizations in 2004 and 58 percent of emergency department visits for the disease between 1999 and 2007. Using Medicaid data from four states with large populations that account for more than one-third of Medicaid program enrollment, we examined the characteristics of those with sickle cell disease. We found instances of mortality rates more than nine times the age-adjusted population average (in Texas, a mortality rate for Medicaid enrollees with SCD of 1.11 percent compared to 0.12 percent overall); rates of disability-related eligibility–which is associated with long-term Medicaid enrollment–of up to 69 percent; and half or more of affected enrollees having (all-cause) hospital stays, emergency department visits, and opioid prescription fills. With gene therapies on the horizon that will spur discussions of treatment coverage, costs, and outcomes for people with sickle cell disease, it is important for relevant stakeholders to understand the affected populations.

Background. An aortoenteric fistula (AEF) describes a communication of the aorta or aortic graft with an adjacent loop of the bowel. Aortic graft erosion is a rare complication of abdominal aortic aneurysm repair. We describe a case of a patient presenting with sepsis from Candida glabrata fungemia secondary to aortoenteric erosion without any symptoms or signs of gastrointestinal bleeding. This is a unique case of Candida glabrata fungemia from aortoenteric graft erosion. Case Summary. This patient is a 75-year-old male with a history of a prior aortobifemoral bypass graft in 2005. He presented with complaints of right paraspinal pain and chills. He had no symptoms of gastrointestinal bleeding or abdominal pain. His white blood cell count was 25,600/mcl (4,000–11,000/mcL) with left shift. The erythrocyte sedimentation rate was 11 mm/hr (0-38 mm/hr), and C-reactive protein was 95.5 mg/L (

Microbial enzymes are increasingly finding applications as therapeutics due to their targeted activity and minimal side effects. Serratiopeptidase, also known as a miracle enzyme, has already proved its potential as an anti-inflammatory, mucolytic, fibrinolytic, analgesic in many studies. A cost effective, bioreactor level production process has been described here comprising of the fed-batch fermentation to produce recombinant serratiopeptidase protein expressed as a fusion construct. High yield of cell mass as well as protein was obtained by the optimization of bioreactor parameters. The downstream solubilization and purification processes were also optimized to achieve maximum yield of pure, active serratiopeptidase protein. A final yield of 2.5 ± 0.764 g L−1 of protein was obtained, having 8382 ± 291 U mg−1 of specific caseinolytic activity. Additionally, a novel, unexpected self-proteolytic activity of the enzyme that cleaves the N-terminal 6× His-SUMO fusion tag along with the enzyme propeptide, thus yielding a mature serratiopeptidase, was also found.

Aim: The aim of the study is to find the correlation between modifiable risk factors such as body mass index (BMI), waist circumference (WC), and smoking with age-related macular degeneration (AMD). Materials and Methods: In this hospital-based, observational, case-control study, 100 patients presenting to our outpatient department over a period of 1 year were included in the study. The selected participants were grouped into the AMD group which included 50 eyes with nonneovascular AMD and neovascular AMD along with 50 eyes as a control group. All participants underwent comprehensive ophthalmological examination along with measurement of weight, height, and WC. Results: Fifty eyes of 50 patients diagnosed to have AMD and 50 eyes of 50 age-matched controls were included in this study. The mean age was 66 years (49–85 years). The mean BMI in patients with neovascular AMD was 27.74 kg/m2 ± 4.93 (n = 19). The mean BMI in patients with nonneovascular AMD was 25.58 kg/m2 ± 2.90 (n = 31), whereas mean BMI in the control group was 23.85 kg/m2 ± 3.31 (n = 50). There was a statistically significant difference between the two AMD groups and the control group with respect to BMI (ANOVA; F = 8.605; DF = 2; P < 0.001). The mean WC in patients with neovascular AMD was 104.63 cm ± 5.78 (n = 19). The mean WC in patients with nonneovascular AMD was 102.6 cm ± 6.1 (n = 31), whereas mean WC in the control group was 91.7 cm ± 6.7 (n = 50), and the difference was statistically significant (ANOVA; F = 42.70; DF = 2; P < 0.001). Statistically significant difference was observed between the smokers belonging to case and control groups (ANOVA; F = 13.967; DF = 2; P < 0.001). Conclusion: A statistically significant correlation exists between BMI, WC, smoking, and the severity of AMD.

Background: Analysis of plasma circulating cell-free DNA integrity (cfDI) has emerged as a promising tool in the diagnosis of malignant vs. benign tumors. There is limited data on the role of cfDI in thyroid cancer. The goal of this study was to analyze cfDI as a biomarker of malignancy in patients with cytologically indeterminate thyroid nodules.Methods: The cfDI was measured in the plasma of patients with cytologically indeterminate thyroid nodules. All patients underwent plasma collection within 24–72 h before surgical treatment for thyroid nodules. Additionally, samples were collected from seven patients via the vein draining the thyroid and peripheral vein during surgery. Quantitative real-time PCR was performed on the isolated cell-free DNA using two different primer sets (115 and 247 bp) to amplify consensus ALU sequences. The cfDI was calculated as the ratio of ALU247 to ALU115.Results: All data are given as median [25th−75th percentile]. The study group consisted of 67 patients with 100 nodules, 80.6% (54/67) women, aged 43 [33-60] years. There was no difference in cfDI between 29 patients with benign nodules (0.49 [0.41–0.59]) and 38 patients with malignant lesions (0.45 [0.36–0.57], p = 0.19). There was no difference in cfDI in the vein draining the thyroid (0.47 [0.24–1.05]) and peripheral vein (0.48 [0.36–0.56], p = 0.44). In comparison to thyroid cancer patients, patients with benign nodules were characterized by significantly higher concentrations of ALU115 (1,064 [529–2,960] vs. 411 [27–1,049] ng/ml; p = 0.002) and ALU247 (548 [276–1,894] vs. 170 [17-540] ng/ml; p = 0.0005), most likely because benign tumors were larger (3, [1.8–4.1 cm]) than malignant lesions (0.7 [0.23–1.45], p < 0.0001). Women had significantly lower cfDI (0.45 [0.27–0.54]) than men (0.56 [0.44–0.8], p = 0.011).Conclusion: The cfDI measured in the vein draining the thyroid is similar to the cfDI measured in the antecubital vein, validating cfDI measurements by peripheral liquid biopsy. Analysis of cfDI needs to be stratified by patients gender. In contrast to its diagnostic utility in aggressive cancers, cfDI has limited utility as a biomarker of malignancy in cytologically indeterminate thyroid nodules.

Retromer is a phylogenetically conserved, multisubunit coat complex that controls endosomal protein trafficking and sorting. Mutations in the retromer gene VPS35 cause late-onset Parkinson disease, suggesting that trafficking defects cause neurodegeneration. Sorting nexins assist retromer to guide cell surface proteins to their assigned destinations, and our interest here is sorting nexin 3 (Snx3). Snx3 binds to membranes via a phox homolog (PX) domain that binds phosphatidylinositol 3-phosphate (PI3P), and in human cells its cargo proteins are the transferrin and Wnt receptors and the divalent metal ion transporter, whereas in yeast the best characterized cargo is the iron permease Ftr1. We recently discovered that α-synuclein inhibits Snx3-retromer recycling of Ftr1 in an unexpected way: α-synuclein, which avidly binds to negatively charged lipids, blocks the association of Snx3 to early endosomes. Here, we discuss mechanisms by which α-synuclein can disrupt Snx3-retromer–mediated recycling.

Objectives: Chloride intracellular channel protein 4 (Clic4) is a ubiquitously expressed protein involved in multiple cellular processes including cell-cycle control, cell differentiation, and apoptosis. Here, we investigated the role of Clic4 in pancreatic β-cell apoptosis. Methods: We used βTC-tet cells and islets from β-cell specific Clic4 knockout mice (βClic4KO) and assessed cytokine-induced apoptosis, Bcl2 family protein expression and stability, and identified Clic4-interacting proteins by co-immunoprecipitation and mass spectrometry analysis. Results: We show that cytokines increased Clic4 expression in βTC-tet cells and in mouse islets and siRNA-mediated silencing of Clic4 expression in βTC-tet cells or its genetic inactivation in islets β-cells, reduced cytokine-induced apoptosis. This was associated with increased expression of Bcl-2 and increased expression and phosphorylation of Bad. Measurement of Bcl-2 and Bad half-lives in βTC-tet cells showed that Clic4 silencing increased the stability of these proteins. In primary islets β-cells, absence of Clic4 expression increased Bcl-2 and Bcl-xL expression as well as expression and phosphorylation of Bad. Mass-spectrometry analysis of proteins co-immunoprecipitated with Clic4 from βTC-tet cells showed no association of Clic4 with Bcl-2 family proteins. However, Clic4 co-purified with proteins from the proteasome suggesting a possible role for Clic4 in regulating protein degradation. Conclusions: Collectively, our data show that Clic4 is a cytokine-induced gene that sensitizes β-cells to apoptosis by reducing the steady state levels of Bcl-2, Bad and phosphorylated Bad.

Introduction: Proximal Femoral Nailing (PFN) in Intertrochanteric Fractures (IF) is becoming the choice of implant due to better biomechanics and prevention of varus collapse associated with Dynamic Hip Screw (DHS). Technical difficulties and implant related complications are yet to be addressed. Aim: To understand the technical difficulties involved in PFN and role of provisional fixation of fracture by Steinmann-pin. Materials and Methods: In this study, 55 patients presented to a tertiary trauma center in India with trochanteric fractures from April 2010 to March 2012 were included and treated with PFN. All patients were followed-up for two years and final outcome assessment included shortening, neck shaft angle and Harris Hip Score was done. Results: In all except one, neck shaft angle greater than 130° was achieved and also maintained in the final follow up (Mean 131.1°). All fractures were united with mean shortening of 3.6 mm and average Harris Hip Score of 91 after two years. There were five complications which included one shortening, two varus collapses, one backed out screws and one reverse Z effect. Conclusion: Though PFN is technically challenging, with proper technique, gives excellent results with negligible varus collapse even in unstable fractures. Three most important technical aspects are achieving good non-varus reduction, inserting nail correctly and accurate placement of lag screws. The technique of provisional fixation of fracture fragments by Steinmann-pin significantly helps in achieving these and reduces the risk of implant failure.

We have been investigating the role that phosphatidylethanolamine (PE) and phosphatidylcholine (PC) content plays in modulating the solubility of the Parkinson's disease protein alpha-synuclein (α-syn) using Saccharomyces cerevisiae and Caenorhabditis elegans. One enzyme that synthesizes PE is the conserved enzyme phosphatidylserine decarboxylase (Psd1/yeast; PSD-1/worms), which is lodged in the inner mitochondrial membrane. We previously found that decreasing the level of PE due to knockdown of Psd1/psd-1 affects the homeostasis of α-syn in vivo. In S. cerevisiae, the co-occurrence of low PE and α-syn in psd1Δ cells triggers mitochondrial defects, stress in the endoplasmic reticulum, misprocessing of glycosylphosphatidylinositol-anchored proteins, and a 3-fold increase in the level of α-syn. The goal of this study was to identify drugs that rescue this phenotype. We screened the Prestwick library of 1121 Food and Drug Administration-approved drugs using psd1Δ + α-syn cells and identified cyclosporin A, meclofenoxate hydrochloride, and sulfaphenazole as putative protective compounds. The protective activity of these drugs was corroborated using C. elegans in which α-syn is expressed specifically in the dopaminergic neurons, with psd-1 depleted by RNAi. Worm populations were examined for dopaminergic neuron survival following psd-1 knockdown. Exposure to cyclosporine, meclofenoxate, and sulfaphenazole significantly enhanced survival at day 7 in α-syn-expressing worm populations whereby 50-55% of the populations displayed normal neurons, compared to only 10-15% of untreated animals. We also found that all three drugs rescued worms expressing α-syn in dopaminergic neurons that were deficient in the phospholipid cardiolipin following cardiolipin synthase (crls-1) depletion by RNAi. We discuss how these drugs might block α-syn pathology in dopaminergic neurons.

Cardiovascular disease is the leading cause of mortality in Cuba. Lifestyle risk factors for coronary heart disease (CHD) in Cubans have not been compared to risk factors in Cuban Americans. Articles spanning the last 20 years were reviewed. The data on Cuban Americans are largely based on the Hispanic Health and Nutrition Examination Survey (HHANES), 1982–1984, while more recent data on epidemiological trends in Cuba are available. The prevalence of obesity and type 2 diabetes mellitus remains greater in Cuban Americans than in Cubans. However, dietary preferences, low physical activity, and tobacco use are contributing to the rising rates of obesity, type 2 diabetes mellitus, and CHD in Cuba, putting Cubans at increased cardiovascular risk. Comprehensive national strategies for cardiovascular prevention that address these modifiable lifestyle risk factors are necessary to address the increasing threat to public health in Cuba.