Class switching is differentially regulated in RBC alloimmunization and vaccination.

BACKGROUND: Studies of human patients have shown that most anti-RBC alloantibodies are IgG1 or IgG3 subclasses, although it is unclear why transfused RBCs preferentially drive these subclasses over others. Though mouse models allow for the mechanistic exploration of class-switching, previous studies of RBC alloimmunization in mice have focused more on the total IgG response than the relative distribution, abundance, or mechanism of IgG subclass generation. Given this major gap, we compared the IgG subclass distribution generated in response to transfused RBCs relative to protein in alum vaccination, and determined the role of STAT6 in their generation. STUDY DESIGN AND METHODS: WT mice were either immunized with Alum/HEL-OVA or transfused with HOD RBCs and levels of anti-HEL IgG subtypes were measured using end-point dilution ELISAs. To study the role of STAT6 in IgG class-switching, we first generated and validated novel STAT6 KO mice using CRISPR/cas9 gene editing. STAT6 KO mice were then transfused with HOD RBCs or immunized with Alum/HEL-OVA, and IgG subclasses were quantified by ELISA. RESULTS: When compared with antibody responses to Alum/HEL-OVA, transfusion of HOD RBCs induced lower levels of IgG1, IgG2b, and IgG2c but similar levels of IgG3. Class switching to most IgG subtypes remained largely unaffected in STAT6 deficient mice in response to HOD RBC transfusion, with the one exception being IgG2b. In contrast, STAT6 deficient mice showed altered levels of all IgG subtypes following Alum vaccination. DISCUSSION: Our results show that anti-RBC class-switching occurs via alternate mechanisms when compared with the well-studied immunogen alum vaccination.

Role of Bilastine in Allergic Rhinitis: A Narrative Review.

Allergic rhinitis (AR) is considered a trivial disease and is often self-treated with over-the-counter drugs and home remedies. However, AR is a contributing risk factor for asthma associated with complications, including chronic cough, eosinophilic esophagitis, and otitis media with effusion. In AR, inflammation is primarily mediated by histamines. Guidelines advise using second-generation oral H1 antihistamines as the primary treatment for AR. Second-generation H1 antihistamines strongly prefer the H1 receptor, limiting their ability to enter the central nervous system. Thus, they have minimal adverse effects. Among these H1 antihistamines, bilastine is highly specific for H1 receptors with a slight affinity for other receptors. It has a rapid and prolonged action, which reduces the need for frequent dosing and has better compliance. In the long term, bilastine is well-tolerated with minimal adverse effects. It is not associated with drug interactions, so dosage adjustment is unnecessary. Bilastine does not penetrate the brain and is nonsedating at 80 mg once daily. The low possibility of drug-drug interactions and pharmacokinetics of bilastine makes it suitable for elderly patients, even with compromised hepatic and renal function, without dose adjustment. This review comprehensively discusses the guidelines and the role of bilastine in treating AR. How to cite this article: Tiwaskar M, Vora A, Tewary K, et al. Role of Bilastine in Allergic Rhinitis: A Narrative Review. J Assoc Physicians India 2023;71(11):58-61.

Role of Ambulatory Glucose Profile in Precision Medicine in Type 2 Diabetes Mellitus.

Assessment of diabetes with daily blood glucose fluctuations including peaks and nadirs forms the crux of the modern management. Use of glycemic variability (GV) as a parameter to assess these fluctuations is emerging. It is important to determine the hyperglycemic and hypoglycemic episodes which are the culprits for increasing glycemic variation. Diabetes mellitus patients follow different clinical trajectories which can be traced by the ambulatory glucose profile (AGP) obtained from flash glucose monitoring system (FGMS). MATERIAL: This comparative observational study enrolled 106 adult (>18 years) type 2 diabetes patients with HbA1c<8%. Patients were divided into two groups (group A & group B) with 53 patients each. Group A included patients on OAD's (oral antidiabetic drug) with insulin and Group B included patients on OAD's without insulin. The patients were put on FGMS for 14 days and their AGP was analysed. Hyperglycemic episodes (level 1- >180 mg/dl, level 2- >250 mg/dl) and hypoglycemic episodes (level 1- 54-70 mg/dl, level 2- <54 mg/dl) were determined between the groups. OBSERVATION: Group A patients had significantly higher (29.99%) total number of hyperglycemic episodes (Level 1+ Level 2) as compared with group B (9.08%) (p <0.0001). Amongst group A, proportion of patients with total number of hyperglycemic episodes was significantly higher in insulin only subgroup (58.11%) followed by insulin +metformin+ 1 OAD (29.14%) & insulin+ metformin (26.82%) (p <0.0001). Amongst group B, total number of hyperglycemic episodes were found to be significantly higher with metformin only subgroup (10.19%) followed by metformin + 1 OAD (9.72%) & metformin + >1 OAD (8.1%) (p<0.0001). Amongst the add on OAD's, sulfonylurea contributed to 61.07% hyperglycemic episodes in group A & 11.63% in group B which was statistically more than DPP-4 inhibitors with 14.91% & 2.84% respectively (p <0.0001). Total number of hypoglycemic episodes seen in group A patients (8.66%) were significantly less as compared with group B (13.27%) (p<0.0001). Sulfonylurea contributed to 7.5% hypoglycemic episodes in group A & 13.2% in group B which was statistically more than DPP-4 inhibitors with 6.49% & 12.35% respectively when added to metformin (p<0.0001). CONCLUSION: Amongst the OAD's used in type 2 diabetes mellitus patients in this study, total number of hyperglycemic and hypoglycemic episodes were found to be more in patients taking sulfonylurea as compared with DPP4 inhibitors when used in combination with metformin with or without insulin.

Transient Elastography of Liver and Serum Hyaluronic Acid Levels In Adult Transfusion Dependent Thalassemia Patients.

Iron overload occurs as a result of multiple blood transfusions and increased iron absorption in thalassemia patients. Iron deposition in liver results in liver stiffness and fibrosis. Non invasive methods including imaging and serum biomarkers have been introduced for assessment of liver fibrosis. We aimed to study liver stiffness using transient elastography and serum hyaluronic acid levels and correlate them with serum ferritin levels in adult transfusion dependent beta thalassemia patients. MATERIAL: 70 transfusion dependent thalassemia patients of age ≥18 years, registered at Thalassemia Day Care Centre were subjected to investigations like CBC, Liver function tests, viral markers, serum ferritin, serum hyaluronic acid levels and transient elastography. Fibrosis indices like FIB-4, AAR and APRI were also calculated. 45 patients had T2*MRI reports with them; which were also included and analysed. Spearman coefficient r was used to test correlations between TE values and serum HA levels with other variables. OBSERVATION: 70 patients (41 male and 29 female) with mean age of 24.09±5.38 years and BMI 20.51 ±3.47 kg/m², were enrolled. Median values of hemoglobin, AST, ALT, TE, serum HA and serum ferritin were, 9.15 g/dl, 42 IU/L, 47.50 IU/L, 9.1 kPa, 284 ng/dl and 1841 ng/ml, respectively . TE values had significant positive correlation with serum ferritin (r=0.5, p < 0.001), ALT (r=0.59, p < 0.001), AST (r=0.58, p< 0.001), APRI (r=0.5, p<0.001) and FIB-4 (p=0.02), respectively and significant negative correlation with T2* MRI (ms) (r= -0.5, p<0.001). No significant correlation of HA was found with any variable. CONCLUSION: Transient elastography can be used as a non expensive, easily accessible and non invasive marker of liver iron overload. Further detailed studies are required to establish the role of serum Hyaluronic acid in thalassemia patients.

Splicing factor DHX15 affects tp53 and mdm2 expression via alternate splicing and promoter usage.

DHX15, a DEAH box containing RNA helicase, is a splicing factor required for the last step of splicing. Recent studies identified a recurrent mutational hotspot, R222G, in DHX15 in ∼ 6% of acute myeloid leukemia (AML) patients that carry the fusion protein RUNX1-RUNX1T1 produced by t (8;21) (q22;q22). Studies using yeast mutants showed that substitution of G for the residue equivalent to R222 leads to loss of its helicase function, suggesting that it is a loss-of-function mutation. To elucidate the role of DHX15 during development, we established the first vertebrate knockout model with CRISPR/Cas9 in zebrafish. Our data showed that dhx15 expression is enriched in the brain, eyes, pectoral fin primordia, liver and intestinal bulb during embryonic development. Dhx15 deficiency leads to pleiotropic morphological phenotypes in homozygous mutant embryos starting at 3 days post fertilization (dpf) that result in lethality by 7 dpf, revealing an essential role during embryonic development. RNA-seq analysis suggested important roles of Dhx15 in chromatin and nucleosome assembly and regulation of the Mdm2-p53 pathway. Interestingly, exons corresponding to the alternate transcriptional start sites for tp53 and mdm2 were preferentially expressed in the mutant embryos, leading to significant upregulation of their alternate isoforms, Δ113p53 (orthologous to Δ133p53 isoform in human) and mdm2-P2 (isoform using distal promoter P2), respectively. We speculate that these alterations in the Mdm2-p53 pathway contribute to the development of AML in patients with t(8;21) and somatically mutated DHX15.

Coagulation Profile and its Correlation with Severity of Liver Dysfunction and Gastrointestinal Bleed in Alcoholic Liver Disease Patients.

INTRODUCTION: The study aimed to determine coagulation factor abnormalities in alcoholic liver disease (ALD) and correlate these with severity of liver dysfunction (Child's class) and gastrointestinal (GI) bleeding. METHODS: 60 patients of ALD (alcohol intake >10years and clinical, biochemical or radiological evidence of chronic liver disease) were included. Patients with Hepatitis B, Hepatitis C, HIV infection, DIC, low platelet count due to other causes, or on drugs which affect coagulation profile were excluded. OBSERVATIONS: Age was 44.42 ± 10.26 years (100% males), 53% in Childs class C. Severity of liver dysfunction showed a significant association (p<0.05) with prolongation of prothrombin time (PT), activated partial thromboplastin time (aPTT) and thrombin time (TT), increasing factor VIII and D-Dimer level, low platelet counts, low protein S and factor VII activity; as well as decreasing fibrinogen levels, protein C and antithrombin (AT) III. GI bleed is associated significantly (p<0.05) with PT >20 sec and decreased plasma fibrinogen levels, while normal protein C, normal AT III, normal factor VII, normal factor VIII, normal TT, increased plasma fibrinogen levels, normal PT and normal platelet count appeared to be protective. CONCLUSIONS: Several coagulation parameters are altered in ALD variably. Alterations in PT, aPTT, TT, factor VIII, D-Dimer, fibrinogen, protein C and AT III levels can be used for grading severity of liver disease. Decreased fibrinogen, protein C activity, AT III activity, factor VII activity, and increased factor VIII activity, are associated with GI bleed.

Ethambutol Optic Neuropathy: Vigilance and Screening, the Keys to Prevent Blindness with the Revised Anti-tuberculous Therapy Regimen.

There has been change in the guidelines for the management of tuberculosis in India. The new guidelines advocate the daily use of Ethambutol for both intensive and continuation phase of the treatment. This may be a matter of concern as increased cumulative dose may lead to increase in incidence of toxic optic neuropathy due to ethambutol. Indian Neuro-Ophthalmology Society has taken cognizance of the issue and has come-up with guidelines for prevention and early detection of the toxic optic neuropathy.

Genotypic diversity in clinical and environmental isolates of Cryptococcus neoformans from India using multilocus microsatellite and multilocus sequence typing.

BACKGROUND: Cryptococcus neoformans is the leading cause of cryptococcal meningitis in HIV/AIDS patients. As infections in humans are predominantly caused by the inhalation of basidiospores from environmental sources, therefore, analysing the population structure of both clinical and environmental populations of C neoformans can increase our understanding of the molecular epidemiology of cryptococcosis. OBJECTIVE: To investigate the genotypic diversity and antifungal susceptibility profile of a large collection of C neoformans isolates (n = 523) from clinical and environmental sources in India between 2001 and 2014. MATERIALS AND METHODS: Cryptococcus neoformans isolates were genotyped by AFLP, microsatellite typing (MLMT) and MLST. In vitro antifungal susceptibility for standard antifungals was undertaken using CLSI M27-A3. RESULTS: All isolates were C neoformans, AFLP1/VNI and exhibited mating-type MATα. MLMT revealed that the majority of isolates belonged to microsatellite cluster (MC) MC3 (49%), followed by MC1 (35%), and the remaining isolates fell in 11 other MC types. Interestingly, two-thirds of clinical isolates were genotype MC3 and only 17% of them were MC1, whereas majority of environmental strains were MC1 (54%) followed by MC3 (16%). Overall, MLST assigned 5 sequence types (STs) among all isolates and ST93 was the most common (n = 76.7%), which was equally distributed in both HIV-positive and HIV-negative patients. Geometric mean MICs revealed that isolates in MC1 were significantly less (P < .05) susceptible to amphotericin B, 5-flucytosine, itraconazole, posaconazole and isavuconazole than isolates in MC3. CONCLUSIONS: The study shows a good correlation between MLMT and MLST genotyping methods. Further, environmental isolates were genetically more diverse than clinical isolates.

The Syndromic Spectrum of COVID-19 and Correlates of Admission Parameters with Severity-outcome Gradients: A Retrospective Study.

BACKGROUND: Clinical and laboratory features of COVID-19 may have regional variations. This study aimed to discern their association with severity of illness and mortality in tertiary setup of Delhi, India. METHODS: Retrospective data of hospitalised COVID-19 patients over 3 months (end March to June 2020) were evaluated for symptom profile, blood investigations and chest radiograph data and classified according to COVID-19 severity and as survivors and non-survivors. RESULTS: Average age (n=182) was 46.1 years, male to female ratio 1.4:1. Fever (51.1%), cough (49.4%) and breathlessness (48.3%) were the commonest symptoms, and frequency of all the three increased with severity of COVID-19. Fever duration, leucocytosis, neutrophilia, elevated blood urea, transaminitis and higher Brixia score on chest X-ray were also more in severe COVID-19 compared to mild and moderate categories. Higher age, more comorbidities, fever, breathlessness and chest pain; longer duration of fever, leucocytosis, neutrophilia, lymphopenia, high neutrophil to lymphocyte ratio, elevated serum urea, creatinine, transaminases and hyperglycemia, and higher radiographic Brixia score were observed in non-survivors compared to survivors. CONCLUSION: Greater prevalence of symptoms (alone and in combination) and derangements in blood biochemistry are seen in severe COVID-19 compared to mild or moderate cases, and also in non-survivors compared to survivors.

COVID-19 and Tuberculosis: A Meeting of Two Pandemics!

Coronavirus disease 2019 (COVID-19), causes serious respiratory illness manifesting as pneumonia, adult respiratory distress syndrome and respiratory failure. Amidst the rising number of cases and deaths, it is imperative to not forget Tuberculosis (TB) which is another pandemic existing since centuries. There could be dire consequences for tuberculosis patients globally especially in low and middle income countries with a high burden of disease and overwhelmed health care systems. Tuberculosis is still the leading infectious killer worldwide, and therefore, it is crucial to reflect on the interaction between the two diseases. Evidence suggests that both COVID-19 and tuberculosis have a synergistic relationship, boosting detrimental effect of each other, disrupting existing health care models, and also worsening the clinical outcomes in terms of morbidity and mortality. This review aims to draw attention towards this pertinent clinical issue, and tries to unravel the intricate relationship between COVID-19 and tuberculosis, as also the role of BCG vaccination to combat the COVID-19 pandemic.

Pancreatic Injury in COVID-19 Patients.

BACKGROUND AND AIM: Coronavirus disease 2019 (COVID 2019) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may cause multisystem dysfunction. We studied pancreatic injury (serum amylase and serum lipase levels) in COVID-19 patients. METHODS: A retrospective study involving 42 COVID-19 patients (diagnosed by real-time PCR) admitted to a tertiary care hospital was conducted. Serum amylase and serum lipase levels were analysed in relation to severity of COVID-19 and mortality. RESULTS: Mean age of patients was 50 ± 16 years, with male to female ratio of 3.7:1. Serum amylase was elevated in 14 patients (33%). Serum lipase was elevated in 7 out of 29 patients (24.1%). Mortality was seen in 18 patients (42.8%). Serum amylase or lipase did not correlate with severity of COVID-19 or its mortality. However, both patients who had high lipase (>3times) died. CONCLUSION: The prevalence of hyperamylasemia in patients of COVID-19 was 33%, while that of elevated lipase was 24.1%. Pancreatic injury failed to show any statistically significant relation to severity or outcome of COVID-19.

API-ISG Consensus Guidelines for Management of Gastrooesophageal Reflux Disease.

Gastroesophageal reflux disease (GERD) is a common problem in the community. The Indian Society of Gastroenterology and Association of Physicians of India have developed this evidence-based practice guideline for management of GERD in adults. A modified Delphi process was used to develop this consensus containing 43 statements, which were generated by electronic voting iteration as well as face-to-face meeting, and review of the supporting literature primarily from India. These statements include 4 on epidemiology, 9 on clinical presentation, 11 on investigations, 18 on treatment (including medical, endoscopic, and surgical modalities), and one on complications of GERD. The statement was regarded as accepted when the proportion of those who voted either to accept completely or with minor reservation was 80% or higher. The prevalence of GERD in large population-based studies in India is approximately 10% and is probably increasing due to lifestyle changes and increase in obesity. The diagnosis of GERD in the community should be mainly based on presence of classical symptoms like heartburn and sour regurgitation, and empiric treatment with a proton pump inhibitor (PPI) or H2 receptor antagonist should be given. All PPIs in equipotent doses are similar in their efficacy in the management of symptoms. Patients in whom symptoms do not respond adequately to PPI are regarded as having PPIrefractory GERD. Invasive investigations should be limited to patients with alarm symptoms and those with refractory GERD.

Effect of inhaled corticosteroids on serum periostin levels in adult patients with mild-moderate asthma.

Background: Periostin is an extracellular matrix protein, and its expression is upregulated in airway epithelial cells in patients with bronchial asthma. Periostin may be a key molecule linked to type 2 T-helper cell inflammation. Inhaled corticosteroids (ICS) may suppress periostin-induced asthmatic inflammation. This study estimated the serum periostin levels in patients with asthma and evaluated the effect of ICS on the levels of periostin, peak expiratory flow rate (PEFR), and quality of life (QOL). Methods: The study design was prospective, open label, and observational. Forty adults with mild-to-moderate bronchial asthma started on ICS and matched healthy controls were enrolled. The patients were evaluated for their serum periostin, PEFR, and QOL by using asthma QOL questionnaire scores at baseline and after 4 weeks. Results: The mean ± standard deviation (SD) serum periostin concentration in patients with asthma was 90.36 ± 19.81 ng/mL, which was significantly higher (p < 0.01) compared with healthy controls (31.88 ± 8.71 ng/mL). ICS treatment for 4 weeks reduced the mean ± SD serum periostin levels to 58.78 ± 14.53 ng/mL. The mean ± SD PEFR increased significantly, from 225.25 ± 60.79 L/min to 292.5 ± 59.18 L/min (p < 0.01), after 4 weeks. Similarly, the mean ± SD when using asthma QOL questionnaire scores significantly increased, from 40.7 ± 7.84 at baseline to 59.33 ± 11.0 on day 28 after ICS therapy (p < 0.01). Conclusion: The serum periostin level, a marker of allergic inflammation and remodeling, increased in bronchial asthma and was reduced with ICS therapy. ICS improved QOL scores and PEFR in patients with asthma.

Profile of Community-Acquired Acute Kidney Injury Defined Using RIFLE Criteria Among Medical In-Patients: A Prospective Descriptive Single Centre Study.

AIM: To determine the proportion of patients who have Acute Kidney Injury (AKI), identify severity of AKI using RIFLE criteria and to identify associated factors with AKI. METHODS: One thousand consecutive medical in-patients were screened for AKI and severity assessed using RIFLE criteria in tertiary care hospital in Northern India. Patients with medical renal disease and obstructive uropathy were excluded. Serum creatinine of all patients were done on days 0, 3, 7 and 14. CKD cases were also excluded. AKI patients were followed at 4 weeks and 3 months. RESULTS: Amongst 1000 patients screened, 65 had AKI. 27(41.5%), 15(23.0%) and 23(35.38%) patients belonged to risk, injury and failure classes of AKI respectively as per RIFLE criteria, and there was incremental risk of mortality (25.92%, 46.33% and 86.95%, p<0.001). In-patients with pneumonia, chronic liver disease and acute gastroenteritis have greater odds of developing AKI, with chronic liver disease having a high mortality (90%). Hypotension (OR- 5.5:1, p=0.002) or leucocytosis at presentation (OR-2.8:1, p<0.001), smokers (OR-2.2:1, p=0.03) and alcoholics (OR-2.5:1, p=0.047) had greater odds of developing AKI. 33(50.7%) patients with AKI died and 27(41.5%) recovered before day 28. Five (7.7%) were seen in class L who had persistently elevated creatinine at day 90 i.e. progressed to ESRD, class E. CONCLUSION: The incidence of AKI among medical in-patients was 6.5%, with an incremental risk of mortality in risk, injury and failure classes. Pneumonia and acute gastroenteritis among infections and chronic liver disease have greater odds of developing AKI. Hypotension, leucocytosis, smoking, alcohol and aetiology are independent risk factors for AKI.

Self-reported Adherence to Medication among Patients with Type II Diabetes Mellitus attending a Tertiary Care Hospital of Delhi.

INTRODUCTION: Diabetes mellitus (DM) is becoming a potential epidemic in India with more than 62 million diagnosed diabetics and an increase of nearly 2 million per year. Poor adherence to medication regimens increases the probability of adverse outcomes in type 2 diabetes patients. Therefore, improving medication adherence is a growing priority to control this epidemic. Hence, this study was conducted to determine the level of adherence to medication in Type II diabetic patients and to study the various factors affecting adherence to medication and the relationship between the severity of diabetes with the adherence categories. METHODS: A cross-sectional study was conducted at medicine outpatient department (OPD) of a tertiary care hospital, New Delhi among 200 type 2 diabetic patients for duration of 2 months using a predesigned and pretested semi-structured interview schedule and diabetes medication adherence was assessed by Morisky's medication adherence scale questionnaire. RESULTS: Out of 200 participants, 32.5% were found to have high adherence while 34.5% and 33% had moderate and low adherence. Factors found to be associated with adherence were age, educational status, longer duration of disease and presence of glucometer. Almost four-fifths of the patients (79.5%) had poor plasma glucose control. CONCLUSION: There is a need to focus on improving adherence among type 2 diabetes patients and strengthening health care systems for regular supply of medicines and provide health education to the patients and their families emphasising the need of adherence to medications.

High-throughput gene targeting and phenotyping in zebrafish using CRISPR/Cas9.

The use of CRISPR/Cas9 as a genome-editing tool in various model organisms has radically changed targeted mutagenesis. Here, we present a high-throughput targeted mutagenesis pipeline using CRISPR/Cas9 technology in zebrafish that will make possible both saturation mutagenesis of the genome and large-scale phenotyping efforts. We describe a cloning-free single-guide RNA (sgRNA) synthesis, coupled with streamlined mutant identification methods utilizing fluorescent PCR and multiplexed, high-throughput sequencing. We report germline transmission data from 162 loci targeting 83 genes in the zebrafish genome, in which we obtained a 99% success rate for generating mutations and an average germline transmission rate of 28%. We verified 678 unique alleles from 58 genes by high-throughput sequencing. We demonstrate that our method can be used for efficient multiplexed gene targeting. We also demonstrate that phenotyping can be done in the F1 generation by inbreeding two injected founder fish, significantly reducing animal husbandry and time. This study compares germline transmission data from CRISPR/Cas9 with those of TALENs and ZFNs and shows that efficiency of CRISPR/Cas9 is sixfold more efficient than other techniques. We show that the majority of published "rules" for efficient sgRNA design do not effectively predict germline transmission rates in zebrafish, with the exception of a GG or GA dinucleotide genomic match at the 5' end of the sgRNA. Finally, we show that predicted off-target mutagenesis is of low concern for in vivo genetic studies.

A multicentre study of antifungal susceptibility patterns among 350 Candida auris isolates (2009-17) in India: role of the ERG11 and FKS1 genes in azole and echinocandin resistance.

Background: Candida auris has emerged globally as an MDR nosocomial pathogen in ICU patients. Objectives: We studied the antifungal susceptibility of C. auris isolates (n = 350) from 10 hospitals in India collected over a period of 8 years. To investigate azole resistance, ERG11 gene sequencing and expression profiling was conducted. In addition, echinocandin resistance linked to mutations in the C. auris FKS1 gene was analysed. Methods: CLSI antifungal susceptibility testing of six azoles, amphotericin B, three echinocandins, terbinafine, 5-flucytosine and nystatin was conducted. Screening for amino acid substitutions in ERG11 and FKS1 was performed. Results: Overall, 90% of C. auris were fluconazole resistant (MICs 32 to ≥64 mg/L) and 2% and 8% were resistant to echinocandins (≥8 mg/L) and amphotericin B (≥2 mg/L), respectively. ERG11 sequences of C. auris exhibited amino acid substitutions Y132 and K143 in 77% (n = 34/44) of strains that were fluconazole resistant whereas WT genotypes, i.e. without substitutions at these positions, were observed in isolates with low fluconazole MICs (1-2 mg/L) suggesting that these substitutions confer a phenotype of resistance to fluconazole similar to that described for Candida albicans. No significant expression of ERG11 was observed, although expression was inducible in vitro with fluconazole exposure. Echinocandin resistance was linked to a novel mutation S639F in FKS1 hot spot region I. Conclusions: Overall, 25% and 13% of isolates were MDR and multi-azole resistant, respectively. The most common resistance combination was azoles and 5-flucytosine in 14% followed by azoles and amphotericin B in 7% and azoles and echinocandins in 2% of isolates.

Quantum Linear System Algorithm for Dense Matrices.

Solving linear systems of equations is a frequently encountered problem in machine learning and optimization. Given a matrix A and a vector b the task is to find the vector x such that Ax=b. We describe a quantum algorithm that achieves a sparsity-independent runtime scaling of O(κ^{2}sqrt[n]polylog(n)/ε) for an n×n dimensional A with bounded spectral norm, where κ denotes the condition number of A, and ε is the desired precision parameter. This amounts to a polynomial improvement over known quantum linear system algorithms when applied to dense matrices, and poses a new state of the art for solving dense linear systems on a quantum computer. Furthermore, an exponential improvement is achievable if the rank of A is polylogarithmic in the matrix dimension. Our algorithm is built upon a singular value estimation subroutine, which makes use of a memory architecture that allows for efficient preparation of quantum states that correspond to the rows of A and the vector of Euclidean norms of the rows of A.

Environmental distribution of Cryptococcus species and some other yeast-like fungi in India.

A study of environmental distribution revealed the occurrence of Cryptococcus neoformans and C. gattii in 9% and 3%, respectively, of 611 samples investigated. C. neoformans showed the highest isolation frequency from tree trunk hollows in Delhi (31%), whereas C. gattii occurred in 12% of the samples in Delhi and 5% in Rajasthan. In addition, Cryptococcus laurentii (=Papiliotrema laurentii), C. rajasthanensis (=Papiliotrema rajasthanensis), C. podzolicus (=Saitozyma podzolica) and C. flavescens (=Papiliotrema flavescens) occurred in 0.5% each. The recovery of C. flavescens and C. podzolicus was new findings for India. One more noteworthy finding was isolation of a new yeast, recently classified as Saitozyma cassiae sp. Novo. The previous strain of this yeast came from tree bark debris in South India. Our isolates came from decayed wood inside a trunk hollow of an Acacia tree in, Bharatpur Bird Sanctuary, Rajasthan. The isolations of novel strains of Cutaneotrichosporon moniliiforme from decayed wood of a Pinus tree was another significant finding. Phenotypically, they differed from T. moniliforme by being encapsulated cells, had melanin-like pigment production and were unable to assimilate d-manitol and d-melezitose. AFLP analysis showed a distinctive banding profile vis-a-vis the reference strains of T. moniliiforme and Cryptotrichosporon anacardii.

Comparison of EUCAST and CLSI Reference Microdilution MICs of Eight Antifungal Compounds for Candida auris and Associated Tentative Epidemiological Cutoff Values.

Candida auris is an emerging multidrug-resistant yeast. So far, all but two susceptibility testing studies have examined ≤50 isolates, mostly with the CLSI method. We investigated CLSI and EUCAST MICs for 123 C. auris isolates and eight antifungals and evaluated various methods for epidemiological cutoff (ECOFF) determinations. MICs (in milligrams per liter) were determined using CLSI method M27-A3, and the EUCAST E.Def 7.3. ANOVA analysis of variance with Bonferroni's multiple-comparison test and Pearson analysis were used on log2 MICs (significance at P values of <0.05). The percent agreement (within ±0 to ±2 2-fold dilutions) between the methods was calculated. ECOFFs were determined visually, statistically (using the ECOFF Finder program and MicDat1.23 software with 95% to 99% endpoints), and via the derivatization method (dECOFFs). The CLSI and EUCAST MIC distributions were wide, with several peaks for all compounds except amphotericin B, suggesting possible acquired resistance. Modal MIC, geometric MIC, MIC50, and MIC90 values were ≤1 2-fold dilutions apart, and no significant differences were found. The quantitative agreement was best for amphotericin B (80%/97% within ±1/±2 dilutions) and lowest for isavuconazole and anidulafungin (58%/76% to 75% within ±1/±2 dilutions). We found that 90.2%/100% of the isolates were amphotericin B susceptible based on CLSI/EUCAST methods, respectively (i.e., with MICs of ≤1 mg/liter), and 100%/97.6% were fluconazole nonsusceptible by CLSI/EUCAST (MICs > 2). The ECOFFs (in milligrams per liter) were similar across the three different methods for itraconazole (ranges for CLSI/EUCAST, 0.25 to 0.5/0.5 to 1), posaconazole (0.125/0.125 to 0.25), amphotericin B (0.25 to 0.5/1 to 2), micafungin (0.25 to 0.5), and anidulafungin (0.25 to 0.5/0.25 to 1). In contrast, the estimated ECOFFs were dependent on the method applied for voriconazole (1 to 32) and isavuconazole (0.125 to 4). CLSI and EUCAST MICs were remarkably similar and confirmed uniform fluconazole resistance and variable acquired resistance to the other agents.