Two NOTCH1 O-fucose sites have opposing functions in mouse retinal angiogenesis.

Previous in vitro studies demonstrated that Fringe glycosylation of the NOTCH1 extracellular domain at O-fucose residues in Epidermal Growth Factor-like Repeats (EGFs) 6 and 8 is a significant contributor to suppression of NOTCH1 activation by JAG1 or enhancement of NOTCH1 activation by DLL1, respectively. In this study, we sought to evaluate the significance of these glycosylation sites in a mammalian model by generating 2 C57BL/6J mouse lines carrying NOTCH1 point mutations, which eliminate O-fucosylation and Fringe activity at EGFs 6 (T232V) or 8 (T311V). We assessed changes to morphology during retinal angiogenesis, a process in which expression of Notch1, Jag1, Dll4, Lfng, Mfng, and Rfng genes coordinate cell-fate decisions to grow vessel networks. In the EGF6 O-fucose mutant (6f/6f) retinas, we observed reduced vessel density and branching, suggesting that this mutant is a Notch1 hypermorph. This finding agrees with prior cell-based studies showing that the 6f mutation increased JAG1 activation of NOTCH1 during co-expression with inhibitory Fringes. Although we predicted that the EGF8 O-fucose mutant (8f/8f) would not complete embryonic development due to the direct involvement of the O-fucose in engaging ligand, the 8f/8f mice were viable and fertile. In the 8f/8f retina, we measured increased vessel density consistent with established Notch1 hypomorphs. Overall, our data support the importance of NOTCH1 O-fucose residues for pathway function and confirms that single O-glycan sites are rich in signaling instructions for mammalian development.

Improving Mobility in Critically Ill Patients in a Tertiary Care ICU: Opportunities and Challenges.

BACKGROUND: Patients in the intensive care unit (ICU) are subjected to prolonged bed rest secondary to critical illness and related therapies. Data suggest that such bed rest can have adverse consequences on the post-discharge quality of life. There is limited data from India on mobilization practices. We undertook a quality improvement (QI) initiative to understand our mobilization practices, identify challenges, and test interventions. MATERIALS AND METHODS: We carried out a three-phase QI project, and the study was conducted in our 24-bedded ICU. Pre-intervention and post-intervention mobilization performance and scores were analyzed. We also recorded data on adverse events and barriers to mobilization. Descriptive statistics were used to report all the results. RESULTS: A total of 140 patients (1,033 patient days) and 207 patients (932 patient days) were included in our initial audit and post-implementation audit, respectively. In pre-implementation, 31.3% of patients were mobilized with an average mobility score of 2 and this improved to 57.9% with average mobility score of 3.4. Additionally, we demonstrated improvements in the mobility scores of our intubated patients (49.8% achieving a mobility score of 3-5 as compared to 16.7%). CONCLUSION: A multidisciplinary approach is feasible and resulted in significant improvements in early mobilization among critically ill adults. HOW TO CITE THIS ARTICLE: Mohan S, Patodia S, Kumaravel S, Venkataraman R, Vijayaraghavan BKT. Improving Mobility in Critically Ill Patients in a Tertiary Care ICU: Opportunities and Challenges. Indian J Crit Care Med 2021;25(1):34-42.

A comparative study of topical tacrolimus and topical triamcinolone acetonide in nodular scabies.

The aim of this study was to compare the efficacy and safety of 0.1% triamcinolone acetonide and 0.03% tacrolimus ointment for the treatment of nodular lesions of scabies. In this double-blind randomized controlled trial, 50 Indian men with postscabeitic persistent nodular lesions (labelled to have nodular scabies (NS)/postscabies prurigo (PSP)) over the external genitalia, were enrolled. They were randomized into two groups to receive either a mid-potency topical corticosteroid (TCS); triamcinolone acetonide 0.1% ointment twice daily, or topical calcineurin inhibitor (CNI); tacrolimus ointment 0.03% twice daily over the nodular lesions for 2 weeks. All patients were followed up on three visits: 1, 2, and 6 weeks, for assessment. Efficacy was evaluated by 5-point range investigator-assessed VAS, and a 4-point severity of pruritus scale (SPS) score. The mean VAS score was higher in triamcinolone group compared to tacrolimus group at both follow-ups, although statistically significant only at 2nd week visit. The fall in mean SPS at both follow-ups was also higher in the steroid group, but the difference was not statistically significant. While, the overall tolerance to either ointment was excellent, mild hypopigmentation, and epidermal atrophy were detected on dermoscopic evaluation of the treated areas at the 2nd week visit in a small subset of patients in the triamcinolone group. The frequency of relapse of lesions and/or pruritus after four weeks of the 2-week therapy was high in both the groups (higher in tarolimus group). Conclusively, although both treatment options provided safe and satisfactory short term improvement, the results from this study suggest an overall higher efficacy of a mid-potency TCS like triamcinolone 0.1% over topical CNI like tacrolimus 0.03%, in the short-term treatment of nodular lesions of NS/PSP. Topical CNIs may be considered for maintenance of effect, if prolonged therapy is warranted.

Synthesis of Silver Nanoparticles Using Buchu Plant Extracts and Their Analgesic Properties.

We herein report for the first time the synthesis and analgesic properties of silver nanoparticles (Ag-NPs) using buchu plant extract. The as-synthesised Ag-NPs at different temperatures were characterised by UV-Vis spectroscopy, Fourier transform infra-red spectroscopy (FTIR) and transmission transform microscopy (TEM) to confirm the formation of silver nanoparticles. Phytochemical screening of the ethanolic extract revealed the presence of glycosides, proteins, tannins, alkaloids, flavonoids and saponins. The absorption spectra showed that the synthesis is temperature and time dependent. The TEM analysis showed that the as-synthesised Ag-NPs are polydispersed and spherical in shape with average particle diameter of 19.95 ± 7.76 nm while the FTIR results confirmed the reduction and capping of the as-synthesised Ag-NPs by the phytochemicals present in the ethanolic extract. The analgesic study indicated that the combined effect of the plant extract and Ag-NPs is more effective in pain management than both the aspirin drug and the extract alone.

Efficacy of Oral Cinacalcet in Non-PTH Nonmalignant Hypercalcemia from Excess 1,25-Dihydroxyvitamin D.

Elevated 1,25-dihydroxyvitamin D (1,25(OH)2D) is a rare cause of non-parathyroid hormone (PTH)-mediated hypercalcemia seen in granulomatous disease, malignancy (most often lymphoma), or genetic mutations. Therapeutic options are limited. We report the case of a 67-year-old White man with nonmalignant, nongranulomatous, 1,25(OH)2D-mediated hypercalcemia treated successfully with cinacalcet. At presentation, he had hypercalcemia, hypercalciuria with recurrent nephrolithiasis, low PTH, elevated 1,25(OH)2D, and normal 25-hydroxyvitamin D. The 1,25(OH)2D levels were inappropriate in the setting of hypercalcemia with low PTH. Evaluations for sarcoidosis, tuberculosis, and malignancy were negative. Genetic testing showed biallelic variants in the CYP24A1 gene. Cinacalcet was trialed and showed normalization of calcium levels. On cinacalcet, biochemical indices showed a slight increase in 1,25(OH)2D and 24-hour urine calcium and mild decrease in PTH. He briefly experienced symptomatic hypocalcemia that resolved after reducing cinacalcet dose. Due to limited symptomatic benefit, he opted to stop cinacalcet. Additional follow-up showed intermittently elevated serum calcium levels after stopping cinacalcet, most recently 10.3 mg/dL. Cinacalcet may be a therapeutic option in nonmalignant, 1,25(OH)2D-mediated hypercalcemia. Further study is necessary to confirm efficacy, understand risks and benefits, and elucidate mechanism(s) of action.

Diabetes-associated Genetic Variation in MTNR1B and Its Effect on Islet Function.

Context: Multiple common genetic variants have been associated with type 2 diabetes, but the mechanism by which they predispose to diabetes is incompletely understood. One such example is variation in MTNR1B, which implicates melatonin and its receptor in the pathogenesis of type 2 diabetes. Objective: To characterize the effect of diabetes-associated genetic variation at rs10830963 in the MTNR1B locus on islet function in people without type 2 diabetes. Design: The association of genetic variation at rs10830963 with glucose, insulin, C-peptide, glucagon, and indices of insulin secretion and action were tested in a cohort of 294 individuals who had previously undergone an oral glucose tolerance test (OGTT). Insulin sensitivity, ß-cell responsivity to glucose, and Disposition Indices were measured using the oral minimal model. Setting: The Clinical Research and Translation Unit at Mayo Clinic, Rochester, MN. Participants: Two cohorts were utilized for this analysis: 1 cohort was recruited on the basis of prior participation in a population-based study in Olmsted County. The other cohort was recruited on the basis of TCF7L2 genotype at rs7903146 from the Mayo Biobank. Intervention: Two-hour, 7-sample OGTT. Main Outcome Measures: Fasting, nadir, and integrated glucagon concentrations. Results: One or 2 copies of the G-allele at rs10830963 were associated with increased postchallenge glucose and glucagon concentrations compared to subjects with the CC genotype. Conclusion: The effects of rs10830963 on glucose homeostasis and predisposition to type 2 diabetes are likely to be partially mediated through changes in α-cell function.

Improving Intravenous and Subcutaneous Insulin Overlap During Treatment of Diabetic Ketoacidosis: A Quality Improvement Project.

Objective: To reduce the frequency of insufficient overlap of intravenous (IV) and subcutaneous (SC) insulin during the treatment of diabetic ketoacidosis (DKA) as a quality improvement project. Patients and Methods: Rates of insufficient IV and SC insulin overlap (< 2-hour overlap, SC insulin given after IV insulin discontinuation, or no SC insulin given after IV insulin discontinuation) were assessed in adults with DKA treated with IV insulin at a large tertiary care referral center in Rochester, Minnesota, from July 1, 2021, to March 15, 2023. After a preintervention analysis period, an electronic medical record-based best practice advisory was introduced to notify hospital providers discontinuing IV insulin if SC long-acting insulin had not been given in the previous 2-6 hours. Demographic characteristics and clinical outcomes before and after intervention were compared. Results: A total of 352 patient encounters were included (251 in the preintervention phase and 101 in the postintervention phase). The rate of insufficient IV to SC insulin overlap decreased from (88 of 251) 35.1% before intervention to (20 of 101) 19.8% after intervention (P=.005). The rate of posttransition hypoglycemia (<70 mg/dL; to convert to mmol/L, multiply by 0.0259) decreased from (27 of 251) 10.7% to (4 of 101) 4% after intervention (P=.04). Rates of posttransition hyperglycemia (>250 mg/dL), rebound DKA, length of hospital stay, and duration of IV insulin therapy were similar before and after intervention. Conclusion: Using quality improvement methodology, the rates of insufficient IV to SC insulin overlap during treatment of DKA in a large tertiary care referral center were measured and reduced through an electronic medical record-based best practice advisory targeting hospital providers.

Determinants for Thyroid Hormone Replacement Therapy in Subclinical Hypothyroidism: A Multicenter Electronic Health Records-Based Study.

Background: The frequency and factors associated with thyroid hormone replacement therapy among patients with subclinical hypothyroidism (SCH) remain uncertain. Methods: In this electronic health records-based observational cohort study, we included adults diagnosed with SCH from four academic centers (the United States and Mexico) from January 1, 2016, to December 31, 2018. We aimed to identify the determinants of thyroid hormone replacement therapy for SCH and the frequency of treated SCH. Results: A total of 796 patients (65.2% women) had SCH, and 165 (20.7%) were treated with thyroid hormone replacement therapy. The treated group was younger [51.0 (standard deviation {SD} 18.3) vs. 55.3 (SD 18.2) years, p = 0.008] and had a higher proportion of women (72.7% vs. 63.2%, p = 0.03) compared with the untreated group. Only 46.7% of patients in the treated group and 65.6% in the untreated group had confirmatory thyroid function tests (TFTs) before the decision to start thyroid hormone replacement therapy was made. There was no difference in the frequency of thyroid autoimmunity evaluation, but a positive thyroid autoimmunity test was more frequent in the treated group compared with the untreated group (48.2% vs. 20.3%, p < 0.001). In a multivariable logistic regression model, female sex (odds ratio [OR] = 1.71 [CI 1.13-2.59], p = 0.01) and index thyrotropin (TSH) level (OR = 1.97 [CI 1.56-2.49], p < 0.001 for every SD [2.75 mIU/L] change) were associated with higher odds of treatment. Conclusions: Among patients with SCH, female sex and index TSH level were associated with higher odds of treatment. Moreover, in our population, the decision to treat or not to treat SCH was often based on only one set of abnormal TFTs, and thyroid autoimmunity assessment was underused.

Mini-Incision Parotidectomy-Our Technique.

Traditional parotidectomy incision was devised by Blair (1912) which was modified by Bailey (1941). Over the years various approaches and techniques have evolved to improve the aesthetic outcome and at the same time for complete disease clearance with reduced complications. In this study, we evaluated the feasibility of our mini-incision parotidectomy technique along with the surgical and quality of life (QOL) outcomes. This prospective case series was conducted at Apollo Hospitals, Bangalore over a period of 2 years (June 2018-August 2020) and includes 20 patients. The surgical outcomes were assessed in terms of feasibility of mini-incision technique with respect to levels of parotid involved and functional outcomes in terms of presence or absence of complications like facial palsy (temporary or permanent), seroma and Frey's syndrome. Patient related quality of life (QOL) outcomes were assessed in terms of post-operative pain score, patient comfort score and cosmetic score by using numerical rating scale-11 (NRS-11). The mini-incision parotidectomy technique is feasible for lesions in all the parotid levels and conversion or lengthening of incision was not needed in any of the cases. 2(10%) patients had temporary facial palsy (House-Brackman grade III) which was recovered within 3 weeks after surgery. One patient (5%) with adenoid cystic carcinoma had permanent facial palsy. Out of 20 patients 2(10%) had seroma and 1(5%) patient presented with Frey's syndrome. Mean post-operative pain score at 0, 6 and 24 h were 4.8, 3.4 and 1.8 out of 10 respectively. Mean comfort score was 9 out of 10 and mean cosmetic score was 9.5 out of 10. Mini-incision parotidectomy technique can render improved functional as well as aesthetic outcomes after parotidectomy without compromising the surgical clearance of the disease process.

Diabetic Ketoacidosis in Pregnancy: Clinical Risk Factors, Presentation, and Outcomes.

CONTEXT: Diabetic ketoacidosis (DKA) in pregnancy is an obstetric emergency with risk of maternofetal death. OBJECTIVE: This work aimed to evaluate DKA events in pregnant women admitted to our inpatient obstetric service, and to examine associated clinical risk factors, presentation, and pregnancy outcomes. METHODS: A retrospective cohort study was conducted at the Mayo Clinic, Rochester, Minnesota, USA, and included women aged 17 to 45 years who were treated for DKA during pregnancy between January 1, 2004 and December 31, 2021. Main outcome measures included maternal and fetal death along with a broad spectrum of maternal and fetal pregnancy outcomes. RESULTS: A total of 71 DKA events were identified in 58 pregnancies among 51 women, 48 (82.8%) of whom had type 1 diabetes. There were no maternal deaths, but fetal demise occurred in 10 (17.2%) pregnancies (6 miscarriages and 4 stillbirths). Maternal social stressors were frequently present (n = 30, 51.0%), and glycemic control was suboptimal (median first trimester glycated hemoglobin A1c = 9.0%). Preeclampsia was diagnosed in 17 (29.3%) pregnancies. Infants born to women with DKA were large for gestational age (n = 16, 33.3%), suffered from neonatal hypoglycemia (n = 29, 60.4%) and required intensive care unit admission (n = 25, 52.1%). CONCLUSION: DKA is associated with a high rate of maternofetal morbidity and fetal loss. Prenatal education strategies for women with diabetes mellitus should include a strong focus on DKA prevention, and clinicians and patients should have a high index of suspicion for DKA in all pregnant women who present with symptoms that could be attributed to this condition.

First Two-Year Observational Exploratory Real Life Clinical Phenotyping, and Societal Impact Study of Parkinson's Disease in Emiratis and Expatriate Population of United Arab Emirates 2019-2021: The EmPark Study.

BACKGROUND: Phenotypic differences in Parkinson's Disease (PD) among locals (Emiratis) and Expatriates (Expats) living in United Arab Emirates have not been described and could be important to unravel local aspects of clinical heterogenicity of PD pointing towards genetic and epigenetic variations. OBJECTIVE: To investigate the range and nature of motor and nonmotor clinical presentations of PD and its impact on time to diagnosis, local service provisions, and quality of life in Emiratis and Expats in UAE, as well as address the presence of current unmet needs on relation to care and etiopathogenesis of PD related to possible genetic and epigenetic factors. METHODS: a cross-sectional one point in time prospective, observational real-life study of 171 patients recruited from PD and Neurology clinics across United Arab Emirates from 2019-2021. Primary outcomes were sociodemographic data, motor and nonmotor symptoms (NMS), including cognition and sleep, and quality of life (QOL) assessments, Results: A total of 171 PD patients (52 Emiratis 119 Expats) were included with mean age (Emiratis 48.5 (13.1) Expats 64.15 (13.1)) and mean disease duration (Emiratis 4.8 (3.2) Expats 6.1 (2.9)). In the Emiratis, there was a significant mean delay in initiating treatment after diagnosis (Emiratis 1.2 (0.9) Expats 1.6 (1.1)), while from a clinical phenotyping aspect, there is a high percentage of akinesia 25 (48.1) or tremor dominant (22 (42.3)) phenotypes as opposed to mixed subtype 67 (56.3) in Expat cohorts; double tremor dominant, especially Emirati females (25%), had a predominant lower limb onset PD. Both Emirati (27.9 (24.0)) and Expat 29.4 (15.6) showed moderate NMS burden and the NMS profile is dominated by Sleep, Fatigue, Mood, Emotional well-being 3.0 (1.1) and Social Stigma 3.5 (0.9) aspects of PDQ8 SI measurements are predicted worse QOL in Emiratis, while lack of social support 2.3 (1.3) impaired QOL in Expat population. Awareness for advanced therapies was low and only 25% of Emiratis were aware of deep brain surgery (DBS), compared to 69% Expats. Only 2% of Emiratis, compared to 32% of Expats, heard of Apomorphine infusion (CSAI), and no (0%) Emiratis were aware of intrajejunal levodopa infusion (IJLI), compared to 13% of expats. CONCLUSION: Our pilot data suggest clinical phenotypic differences in presentation of PD in Emiratis population of UAE compared to expats. Worryingly, the data also show delayed treatment initiation, as well as widespread lack of knowledge of advanced therapies in the Emirati population.

Observations on Citrobacter species from a tertiary care health center with special reference to multi-drug resistance and presence of CTX-M gene.

BACKGROUND: Citrobacter is an important nosocomial pathogen and its multi-drug resistant (MDR) isolates are increasingly being reported across the globe. They are known to produce extended spectrum beta lactamase (ESBL) and harbor CTX-M gene. OBJECTIVE: The aim was to isolate Citrobacter sp. from clinical specimens, analyze their MDR status and look for the presence of CTX-M gene. MATERIALS AND METHODS: In this prospective study, Citrobacter isolates positive for ESBL on screening, were confirmed by combined disc method along with minimum inhibitory concentration (MIC) for cefotaxime. In selected cefotaxime resistant isolates, multiplex polymerase chain reaction was done for blaCTX-M gene. RESULTS: Of 146 Citrobacter sp. isolated, most (73%) were from admitted patients and hospital stay of >72 h and prior antibiotic intake were the most common associated factors. Maximum isolates were from pus (41.1%). Citrobacter freundii was the commonest species (49%) followed by Citrobacter koseri (28%); 79 were ESBL producers. Seventy were cefotaxime resistant as shown by MIC. blaCTX-M gene was detected in 15/40 of these isolates, all belonged to CTX-M group 1. CONCLUSION: Overall incidence of Citrobacter in our setup is low, but they were mostly MDR, and ESBL production was high, which is a cause of concern. blaCTX-M gene detection is important because of its rapid transmission to other bacterial species.

Completely green synthesis of dextrose reduced silver nanoparticles, its antimicrobial and sensing properties.

We herein report the green synthesis of highly monodispersed, water soluble, stable and smaller sized dextrose reduced gelatin capped-silver nanoparticles (Ag-NPs) via an eco-friendly, completely green method. The synthesis involves the use of silver nitrate, gelatin, dextrose and water as the silver precursor, stabilizing agent, reducing agent and solvent respectively. By varying the reaction time, the temporal evolution of the growth, optical, antimicrobial and sensing properties of the as-synthesised Ag-NPs were investigated. The nanoparticles were characterized using UV-vis absorption spectroscopy, Fourier transform infra-red spectroscopy (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM) and high resolution transmission electron microscopy (HR-TEM). The absorption maxima of the as-synthesized materials at different reaction time showed characteristic silver surface plasmon resonance (SPR) peak. The as-synthesised Ag-NPs show better antibacterial efficacy than the antibiotics; ciproflaxin and imipenem against Pseudomonas aeruginosa with minimum inhibition concentration (MIC) of 6 µg/mL, and better efficacy than imipenem against Escherichia coli with MIC of 10 µg/mL. The minimum bactericidal concentration (MBC) of the as-synthesised Ag-NPs is 12.5 µg/mL. The sensitivity of the dextrose reduced gelatin-capped Ag-NPs towards hydrogen peroxide indicated that the sensor has a very good sensitivity and a linear response over wide concentration range of 10(-1)-10(-6)M H2O2.