Trends of humoral immune responses to heterologous antigenic exposure due to vaccination & omicron SARS-CoV-2 infection: Implications for boosting

Background & objectives: Vaccination and natural infection can both augment the immune responses against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but how omicron infection has affected the vaccine-induced and hybrid immunity is not well studied in Indian population. The present study was aimed to assess the durability and change in responses of humoral immunity with age, prior natural infection, vaccine type and duration with a minimum gap of six months post-two doses with either ChAdOx1 nCov-19 or BBV152 prior- and post-emergence of the omicron variant. Methods: A total of 1300 participants were included in this observational study between November 2021 and May 2022. Participants had completed at least six months after vaccination (2 doses) with either ChAdOx1 nCoV-19 or an inactivated whole virus vaccine BBV152. They were grouped according to their age (? or ?60 yr) and prior exposure of SARS-CoV-2 infection. Five hundred and sixteen of these participants were followed up after emergence of the Omicron variant. The main outcome was durability and augmentation of the humoral immune response as determined by anti-receptor-binding domain (RBD) immunoglobulin G (IgG) concentrations, anti-nucleocapsid antibodies and anti-omicron RBD antibodies. Live virus neutralization assay was conducted for neutralizing antibodies against four variants – ancestral, delta and omicron and omicron sublineage BA.5. Results: Before the omicron surge, serum anti-RBD IgG antibodies were detected in 87 per cent participants after a median gap of eight months from the second vaccine dose, with a median titre of 114 [interquartile range (IQR) 32, 302] BAU/ml. The levels increased to 594 (252, 1230) BAU/ml post- omicron surge (P<0.001) with 97 per cent participants having detectable antibodies, although only 40 had symptomatic infection during the omicron surge irrespective of vaccine type and previous history of infection. Those with prior natural infection and vaccination had higher anti-RBD IgG titre at baseline, which increased further [352 (IQR 131, 869) to 816 (IQR 383, 2001) BAU/ml] (P<0.001). The antibody levels remained elevated after a mean time gap of 10 months, although there was a decline of 41 per cent. The geometric mean titre was 452.54, 172.80, 83.1 and 76.99 against the ancestral, delta, omicron and omicron BA.5 variants in the live virus neutralization assay. Interpretation & conclusions: Anti-RBD IgG antibodies were detected in 85 per cent of participants after a median gap of eight months following the second vaccine dose. Omicron infection probably resulted in a substantial proportion of asymptomatic infection in the first four months in our study population and boosted the vaccine-induced humoral immune response, which declined but still remained durable over 10 months

To evaluate the functional and anatomical outcomes for autologous retinal autograft with Finesse™ Flex Loop for failed macular holes

Purpose: To evaluate the functional and anatomical outcomes for autologous retinal autograft with Finesse™ Flex Loop for failed macular holes. Methods: This is a retrospective study analyzing medical records of consecutive patients with refractory macular hole (at least 1 prior surgery) and eyes with retinal detachment with coexisting macular holes (MH). Optical coherence tomography (OCT) of the macula was performed before and after surgery. The primary study outcome evaluated were the functional and anatomic macular hole closure, and secondary outcomes were improvement in visual acuity and restoration of the outer retinal bands, external limiting membrane, and ellipsoid zone in eyes with acquisition of autologous retinal transplant using Finesse Loop. Results: The study included eight eyes of eight patients; retinal autograft was performed in six (75.0%) and autologous retinal transplantation (ART) with rhegmatogenous retinal detachment (RRD) was performed in two (25.0%) eyes. The average MH basal diameter in the study was 1310.88 ± 138.63 ?m. The successful hole closure rate was observed to be 75% (6 eyes) and 100% retinal reattachment was observed in ART with RRD. Statistically significant (P = 0.001) improvement was noted for preop and postop visual acuity gain for ART acquisition and postop macular hole closure. The restoration of the external limiting membrane (ELM) and ellipsoid zone (EZ) was observed in 37.5% (3 eyes) of patients. Conclusion: Finesse™ Flex Loop can be used to harvest retinal tissue and it provides good anatomical and functional outcomes for failed macular hole.

Comparison of patient satisfaction with red-free (green) versus yellow light using binocular indirect ophthalmoscope for retinal examination

Purpose: Comparison of patient satisfaction with red?free (green) versus yellow light using binocular indirect ophthalmoscope for retinal examination. Methods: This is an observational questionnaire?based study of 100 myopes in the age group of 18–40 years coming for a routine check?up or for refractive surgery workup. The examination was done using an indirect ophthalmoscope and a 20D lens with green or yellow light and was assigned in two groups randomly using the coin toss method, following which, a questionnaire was used to assess the following parameters: a) level of comfort, b) any complaints of discomfort during examination, d) preference of the used light source in future, e) grading of discomfort on a linear scale, and f) patient cooperation and duration of examination. Results: Patients were randomized for observation with IDO using either green light (n = 55) or yellow light (n = 45) filter. In the study, 46 patients (83.6%) were very comfortable and only 9 patients (16.4%) experienced mild discomfort when using red?free (green) light, while only 3 patients (6.7%) were very comfortable and 31 (68.9%) had mild discomfort when using yellow light. The complaints of watering with yellow and green light were noted in 36 patients (80.0%) and 15 patients (27.3%), pain in 13 patients (28.9%) and 3 patients (5.5%), light sensitivity in 29 patients (64.4%) and 4 patients (7.3%), respectively, all being significantly more in yellow light category (P < 0.001). The time of examination was significantly more in yellow light category with 83 ± 10.75 seconds (P < 0.001). Conclusion: Retinal examination using the green filter of indirect ophthalmoscope is more comfortable in examining the patients as compared to routine yellow light with decreased number of complaints, lesser examination time, and better patient cooperation.

Primary retroperitoneal extraovarian granulosa cell tumor

Extraovarian granulosa cell tumors (GCTs) develop from ectopic gonadal tissue situated along the embryonal route of the genital ridge. Primary retroperitoneal tumors are extremely rare, with an incidence of 02% -06% and 80-85% probability of malignancy. Only eight such case reports have been published previously. We herein, report a rare case of extraovarian retroperitoneal GCT in a 55-year-old woman who presented with intermittent left lumbar region pain of one-year duration. She had a history of hysterectomy and bilateral salpingo-oophorectomy 8 years ago for uterine leiomyoma. Laparotomy revealed a retroperitoneal mass measuring 8cm x 10cm x 20cm in size, solid cystic with areas of necrosis and hemorrhage. The gross features, classical histopathology, and positive immunostaining of the retroperitoneal mass with inhibin, calretinin, PR, WT1 and immunonegativity for EMA were characteristic of adult-type GCT. Excluding any previous history of primary ovarian GCT in this patient, a de-novo retroperitoneal diagnosis was established.

Tuberculosis Of Thyroid: A Case Report

Tuberculosis of thyroid is extremely rare, even in regions with high prevalence of tuberculosis. During last century case reports of TTB appeared in journals across the world and the total number of such cases is about a hundred; many of which are post–mortem. We are presenting one such case of Tuberculosis of thyroid and discussing its management.

Oxidative Stress is Independent Factor for End-stage Renal Disease in Type 2 Diabetes Mellitus Patients

An imbalance between oxidant and antioxidants is thought to precede the development of renal lesions. The aim of present study is to determine the relationship between oxidative markers and severity of microalbuminuria in patients with type 2 diabetes mellitus (T2DM). Atotal of 100 T2DM patients (50 males and 50 females) participated in this study. They were screened for microalbuminuria along with oxidative status in patients. Microalbuminuria was detected by measuring the albumin to creatinine ratio (ACR) in urine samples. Patients were divided into two groups; normoalbuminuria (n=36) and microalbuminuria (n=64) as per the ACR levels. No difference in the groups was observed in terms of age, sex, glycated hemoglobin (HbA1c) and blood pressure. The level of oxidative stress was significantly higher in microalbuminuria group of T2DM patients. A significant correlation was observed between ACR and lipid peroxidation (MDA) levels. We conclude that oxidative stress is one of the important mediators of end-stage renal disease (ESRD).

Early detection of multidrug resistant (MDR) Mycobacterium tuberculosis in a single tube with in-house designed fluorescence resonance energy transfer (FRET) probes using real-time PCR.

Rapid and correct diagnosis is crucial for the management of multidrug resistance (MDR) in Mycobacterium tuberculosis (MTB). The present study aims at rapid diagnosis for identification of multidrug resistance tuberculosis (MDR-TB) using real-time PCR. FRET hybridization probes targeting most prominent four selected codons for rpoB526 and 531 and for katG314 and 315 genes were designed and evaluated on 143 clinical MTB isolates and paired sputa for rapid detection of MDR-TB. The results of real-time PCR were compared with gold standard L-J proportion method and further validated by DNA sequencing. Of the 143 MTB positive cultures, 85 and 58 isolates were found to be ‘MDR’ and ‘pan susceptible’, respectively by proportion L-J method. The sensitivity of real-time PCR for the detection of rifampicin (RIF) and isoniazid (INH) were 85.88 and 94.11%, respectively, and the specificity of method was found to be 98.27%. DNA sequencing of 31 MTB isolates having distinct melting temperature (Tm) as compared to the standard drug susceptible H37Rv strain showed 100% concordance with real-time PCR results. DNA sequencing revealed the mutations at Ser531Leu, His526Asp of rpoB gene and Ser315Thr, Thr314Pro of katG gene in RIF and INH resistance cases. This real-time PCR assay that targets limited number of loci in a selected range ensures direct and rapid detection of MDR-TB in Indian settings. However, future studies for revalidation as well as refinement are required to break the limitations of MDR-TB detection.

Genetic diversity of Mycobacterium tuberculosis isolates from central India.

Background & objectives: There is a paucity of data available on genetic biodiversity of Mycobacterium tuberculosis isolates from central India. The present study was carried out on isolates of M. tuberculosis cultured from diagnostic clinical samples of patients from Bhopal, central India, using spoligotyping as a method of molecular typing. Methods: DNA was extracted from 340 isolates of M. tuberculosis from culture, confirmed as M. tuberculosis by molecular and biochemical methods and subjected to spoligotyping. The results were compared with the international SITVIT2 database. Results: Sixty five different spoligo international type (SIT) patterns were observed. A total of 239 (70.3%) isolates could be clustered into 25 SITs. The Central Asian (CAS) and East African Indian (EAI) families were found to be the two major circulating families in this region. SIT26/CAS1_DEL was identified as the most predominant type, followed by SIT11/EAI3_IND and SIT288/CAS2. Forty (11.8%) unique (non-clustered) and 61 (17.9%) orphan isolates were identified in the study. There was no significant association of clustering with clinical and demographic characteristics of patients. Interpretation & conclusions: Well established SITs were found to be predominant in our study. SIT26/CAS1_DEL was the most predominant type. However, the occurrence of a substantial number of orphan isolates may indicate the presence of active spatial and temporal evolutionary dynamics within the isolates of M. tuberculosis.

Successful management of difficult-to-treat irritable bowel syndrome incorporating a psychological approach.

Irritable bowel syndrome is a common gastrointestinal condition with underlying psychological factors. Its management can be challenging, sometimes necessitating a multidisciplinary team of gastroenterologists, psychiatrists and clinical psychologists. Non-pharmacological interventions are gaining attention for the management of chronic irritable bowel syndrome. We present a difficult-to-treat case of chronic irritable bowel syndrome, which was managed successfully with psychological interventions. Natl Med J India 2015;28:188–9

Targeting rubella for elimination.

Rubella is an acute, usually mild viral disease. However, when rubella infection occurs just before conception or during the first 8-10 weeks of gestation, it causes multiple fetal defects in up to 90% of cases, known as Congenital Rubella Syndrome (CRS). It may result in fetal wastage, stillbirths and sensorineural hearing deficit up to 20 weeks of gestation. Rubella vaccine (RA 27/3) is highly effective and has resulted in elimination of rubella and CRS from the western hemisphere and several European countries. Review of several studies documents the duration of protection over 10-21 years following one dose of RA27/3 vaccination, and persistent seropositivity in over 95% cases. Studies in India show seronegativity to rubella among adolescent girls to vary from 10% to 36%. Although due to early age of infection resulting in protection in the reproductive age group, incidence of rubella in India is not very high. However, due to severity of CRS coupled with introduction of RCV in private sector and in some of the states which is likely to lead to sub-optimal coverage and resulting higher risk of rubella during pregnancy in the coming decades, it is imperative to adopt the goal of rubella elimination. As in order to control measles, the country has adopted strategy of delivering second dose of measles through measles campaigns covering children 9 months to 10 years of age in 14 states, it is recommended to synergize efforts for elimination of rubella with these campaigns by replacing measles vaccine by MR or MMR vaccine. Other states which are to give second dose of measles through routine immunization will also have to adopt campaign mode in order to eliminate rubella from the country over 10-20 years. Subsequently, measles vaccine can be replaced by MR or MMR vaccine in the national schedule.

Whole cell & culture filtrate proteins from prevalent genotypes of Mycobacterium tuberculosis provoke better antibody & T cell response than laboratory strain H37Rv.

Background & objectives: The immune responses to different antigens of Mycobacterium tuberculosis H37Rv vary from patient to patient with tuberculosis (TB). Therefore, significant difference might be documented between the H37Rv with long histories of passages and recent clinical isolates of M. tuberculosis. In the present study, immune response of TB patients and healthy controls against 39 clinical M. tuberculosis isolates was correlated with laboratory strain H37Rv. Methods: The antibody response was studied coating whole cell extracts and culture filtrate proteins of M. tuberculosis isolates and laboratory strain H37Rv by enzyme linked immunosorbent assay (ELISA). Lymphoproliferation was studied by incorporation of tritiated thymidine and cytokines (IFN-γ and IL-4) by using commercially available kits. Results: Sero-reactivity to whole cell extract (WCE) of 11 clinical isolates was higher with pooled serum and individual's serum from tuberculosis patients showed significant reactivity (P<0.05) to ten of these isolates using ELISA. Of the WCE of 39 clinical isolates, 10 were found to be potent inducer of lymphoproliferation as well as cytokine secretion (P<0.05) in peripheral blood mononuclear cells from PPD+ healthy controls. Six culture filtrate proteins (CFPs) from these selected clinical isolates were also better inducers of antibody and T-cell response. Interpretation & conclusion: Overall, our results revealed that the clinical isolates belonging to prevalent genotypes; CAS1_Del (ST-26), East African-Indian (ST-11) and Beijing family (ST-1) induced better antibody and T cell responses compared to H37Rv laboratory strain. Further studies need to be done to purify and identify the dominant protein (s) using whole cell extract and culture filtrates from these immunologically relevant clinical M. tuberculosis isolates, which will be worthwhile to find out pathogenic factors, potential diagnostic markers and protective molecules for tuberculosis.

Typhoid vaccine: A case for inclusion in national program.

Typhoid has been reported to be a common and significant cause of morbidity in pre-school and school-age children in the endemic countries like India. The incidence of typhoid has been reported to be as high as 27.3 per 1000 person-years in children less than 5 years of age. Serious complications occur in about 10% of cases requiring hospitalization. The mean cost of treatment per episode of blood culture-confirmed typhoid fever has been calculated as INR 3,597 (1996 prices) in an outdoor setting, whereas in case of hospitalization, the cost of illness increases by several folds (INR 18,131). Vi polysaccharide vaccine is safe, efficacious and affordable for use as a cost-effective public health tool to protect children from typhoid and related complications, when given at 2 and 5 years of age as a part of National Immunization Schedule.

Molecular typing of Mycobacterium leprae strains from northern India using short tandem repeats.

Background & objectives: Due to the inability to cultivate Mycobacterium leprae in vitro and most cases being paucibacillary, it has been difficult to apply classical genotyping methods to this organism. The objective of this study was therefore, to analyze the diversity among M. leprae strains from Uttar Pradesh, north India, by targeting ten short tandem repeats (STRs) as molecular markers. Methods: Ninety specimens including 20 biopsies and 70 slit scrappings were collected in TE buffer from leprosy patients, who attended the OPD of National JALMA Institute for Leprosy and Other Mycobacterial Diseases, Tajganj, Agra, and from villages of Model Rural Health Research Unit (MRHRU) at Ghatampur, Kanpur, Uttar Pradesh. DNA was extracted from these specimens and ten STRs loci were amplified by using published and in-house designed primers. The copy numbers were determined by electrophoretic mobility as well as sequence analysis. Phylogenetic analysis was done on variable number of tandem repeats (VNTRs) data sets using start software. Results: Diversity was observed in the cross-sectional survey of isolates obtained from 90 patients. Allelic index for different loci was found to vary from 0.7 to 0.8 except for rpoT for which allelic index was 0.186. Similarity in fingerprinting profiles observed in specimens from the cases from same house or nearby locations indicated a possible common source of infection. Such analysis was also found to be useful in discriminating the relapse from possible reinfection. Interpretation & conclusions: This study led to identification of STRs eliciting polymorphism in north Indian strains of M. leprae. The data suggest that these STRs can be used to study the sources and transmission chain in leprosy, which could be very important in monitoring of the disease dynamics in high endemic foci.

Problems related to menstruation amongst adolescent girls.

OBJECTIVE: To study the types and frequency of problems related to menstruation in adolescent girls and the effect of these problems on daily routine. METHODS: Girls in the age group 13-19 years who had had menarche for at least one year at the time of study. 198 adolescent girls have been studied. Data was collected by personal interviews on a pre-tested, semi-structured questionnaire. The questions covered menstrual problems, regularity of menses in last three cycles of menstruation and the effect of these problems on the daily routine. Analysis was done using SPSS version 12. Percentages were calculated for drawing inferences. RESULTS: More than a third (35.9%) of the study subjects were in the age group 13-15 years followed by 17-19 years, 15-17 years respectively. Mean age of study participants was calculated to be 16.2 years. Dysmenorrhea (67.2%) was the commonest problem and (63.1%) had one or the other symptoms of Pre-menstrual syndrome (PMS). Other related problems were present in 55.1% of study subjects. Daily routine of 60% girls was affected due to prolonged bed rest, missed social activities/commitments, disturbed sleep and decreased appetite. 17.24% had to miss a class and 25% had to abstain from work. Mothers and friends were the most common source of information on the issue. CONCLUSION: Screen adolescent girls for menstruation related problems and provide them with counseling services and relevant information on possible treatment options. Besides, there is a need to emphasize on designing menstrual health programmes for adolescents.

Analysis of time-dependent recovery from beryllium toxicity following chelation therapy and antioxidant supplementation.

Efforts have been made to minimize the toxic effect caused by beryllium. Adult cyclic rats of Sprague Dawley strain were administered a bolus dose of 50mg/kg beryllium nitrate intramuscularly. The chelation therapy with glutathione (GSH), dimercapto propane sulfonic acid (DMPS)+ selenium (Se) and D-Penicillamine (DPA) + Se was given for 3 days followed by a rest of 1,3 and 7 days respectively. The results revealed a significant fall in the blood sugar level, serum alkaline phosphatase activity, serum proteins. A significant rise in the transaminases i.e. aspartate aminotranferase and alanine aminotranferase pattern is indicative of leakage of enzymes from liver resulting in alterations in the cell permeability. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. Results of the distribution studies by atomic absorption spectrophotometry reveal an increased concentration of beryllium in liver and kidney followed by lung and uterus. The relative ability of 3 chelating agents to act as antagonists for acute beryllium poisoning have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration-dependent during the entire period being highly significant after 7 days rest. From the biochemical assays, and distribution studies it can be assumed that DPA+Se was the most effective therapeutic agent followed by DMPS+Se and GSH. Thus it can be concluded that DPA+Se is a better therapeutic agent as compared to DMPS+Se and GSH.

Role of chelating agents and antioxidants in beryllium induced toxicity.

The present study was conducted to evaluate the therapeutic effectiveness of chelating agents [glutathione, 2,3 dimercapto propane sulfonic acid (DMPS) and D-penicillamine (DPA)] in combination with antioxidant (sodium selenite) in beryllium induced toxicity in female rats. A bolus dose of 50mg/kg-beryllium nitrate was administered singly followed by chelation therapy with GSH, DMPS + Se and DPA + Se at various durations of 1,3 and 7 days respectively. Results revealed a significant fall in the glycogen content, whereas, a marginal fall in the protein was also observed. The enzymatic activity of alkaline phosphatase and adenosine triphosphatase was depleted; on the contrary, there was a significant rise in the acid phosphatase and glucose-6-phosphatase pattern. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. The distribution of the metal by atomic absorption spectrophotometry revealed an increased concentration of beryllium in liver and kidney, followed by lung and uterus. The relative ability of three chelating agents to act as antagonists, for acute beryllium poisoning, have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration dependent during the entire period being highly significant at 7 days regimen. Biochemical and distribution studies reveal that DPA + Se was the most effective therapeutic agent followed by DMPS + Se and GSH.