Development of a novel serological assay for the detection of mpox infection in vaccinated populations.

In 2022 the World Health Organization declared a Public Health Emergency for an outbreak of mpox, the zoonotic Orthopoxvirus (OPV) affecting at least 104 nonendemic locations worldwide. Serologic detection of mpox infection is problematic, however, due to considerable antigenic and serologic cross-reactivity among OPVs and smallpox-vaccinated individuals. In this report, we developed a high-throughput multiplex microsphere immunoassay using a combination of mpox-specific peptides and cross-reactive OPV proteins that results in the specific serologic detection of mpox infection with 93% sensitivity and 98% specificity. The New York State Non-Vaccinia Orthopoxvirus Microsphere Immunoassay is an important tool to detect subclinical mpox infection and understand the extent of mpox spread in the community through retrospective analysis.

Development of a Novel Serological Assay for the Detection of Mpox Infection in Vaccinated Populations.

In 2022 the World Health Organization declared a Public Health Emergency for an outbreak of mpox, the zoonotic Orthopoxvirus (OPV) affecting at least 103 non-endemic locations world-wide. Serologic detection of mpox infection is problematic, however, due to considerable antigenic and serologic cross-reactivity among OPVs and smallpox-vaccinated individuals. In this report, we developed a high-throughput multiplex microsphere immunoassay (MIA) using a combination of mpox-specific peptides and cross-reactive OPV proteins that results in the specific serologic detection of mpox infection with 93% sensitivity and 98% specificity. The New York State Non-Vaccinia Orthopoxvirus Microsphere Immunoassay is an important diagnostic tool to detect subclinical mpox infection and understand the extent of mpox spread in the community through retrospective analysis.

Opportunities and challenges for the development of "core outcome sets" in neuro-oncology.

Core Outcome Sets (COS) define minimum outcomes to be measured and reported in clinical effectiveness trials for a particular health condition/health area. Despite recognition as critical to clinical research design for other health areas, none have been developed for neuro-oncology. COS development projects should carefully consider: scope (how the COS should be used), stakeholders involved in development (including patients as both research partners and participants), and consensus methodologies used (typically a Delphi survey and consensus meeting), as well as dissemination plans. Developing COS for neuro-oncology is potentially challenging due to extensive tumor subclassification (including molecular stratification), different symptoms related to anatomical tumor location, and variation in treatment options. Development of a COS specific to tumor subtype, in a specific location, for a particular intervention may be too narrow and would be unlikely to be used. Equally, a COS that is applicable across a wider area of neuro-oncology may be too broad and therefore lack specificity. This review describes why and how a COS may be developed, and discusses challenges for their development, specific to neuro-oncology. The COS under development are briefly described, including: adult glioma, incidental/untreated meningioma, meningioma requiring intervention, and adverse events from surgical intervention for pediatric brain tumors.

An Association of Pathogens and Biofilms with Alzheimer's Disease.

As one of the leading causes of dementia, Alzheimer's disease (AD) is a condition in which individuals experience progressive cognitive decline. Although it is known that beta-amyloid (Aß) deposits and neurofibrillary tangles (NFT) of tau fibrils are hallmark characteristics of AD, the exact causes of these pathologies are still mostly unknown. Evidence that infectious diseases may cause AD pathology has been accumulating for decades. The association between microbial pathogens and AD is widely studied, and there are noticeable correlations between some bacterial species and AD pathologies, especially spirochetes and some of the oral microbes. Borrelia burgdorferi has been seen to correlate with Aß plaques and NFTs in infected cells. Because of the evidence of spirochetes in AD patients, Treponema pallidum and other oral treponemes are speculated to be a potential cause of AD. T. pallidum has been seen to form aggregates in the brain when the disease disseminates to the brain that closely resemble the Aß plaques of AD patients. This review examines the evidence as to whether pathogens could be the cause of AD and its pathology. It offers novel speculations that treponemes may be able to induce or correlate with Alzheimer's disease.

Exposure to Repeated Mini-objective Structured Practical Examinations as A Formative Assessment Tool to the Students and Faculty in Physiology Practicals.

The study aimed to look at the influence of mini-Objective Structured Practical Examinations (OSPEs) as a formative assessment tool for students and teachers. All first year MBBS students of 2013-14 batch of St. John's Medical College (n=60) went through four mini-OSPE sessions consisting of 5 sets of 6 stations each during 3 months of hematology practicals and final OSPE after 2 months. Feedback of the students and faculty was obtained. The students were categorized into tertiles based on the cumulative scores obtained in the first three-OSPEs and final-OSPE session and were significantly associated from Chi square test. [c2(4, N=54) =15.2, p=0.004]. Low performers with high probability of performing poorly were recognized early in the term. High performers largely remained so and mid performers showed highly variable performance. The feedback obtained supported the role of OSPE as a formative assessment tool. Thus, OSPE could be used as a formative tool for students and teachers.

Effect of Curcumin in Experimental Peritonitis.

Despite medical advancements, the inflammatory cascade and oxidative stress worsen the prognosis in most cases of peritonitis. Curcumin has emerged as a potential antioxidant and anti-inflammatory agent in few of the acute inflammatory and infective conditions. We examined the effect of intraperitoneal injection of curcumin in endotoxin-induced peritonitis in rats. The blood and peritoneal fluid samples were collected at 3 and 24 h following the induction of peritonitis. Animals were sacrificed at 24 h and the organs preserved. The histopathological report of lung, liver, and intestines in the curcumin-treated rats showed maintenance of tissue architecture to a large extent compared to the control group which showed massive congestion, hemorrhage, and necrosis. The blood and peritoneal fluid total count and differential neutrophil counts were significantly higher at 24 h of induction of peritonitis. Serum amyloid assay and lipid peroxidation were significantly lower, and myeloperoxidase assay was higher in the curcumin-treated group at the end of 24 h; thus, curcumin probably demonstrated a neutrophil-mediated immunopotentiation and anti-inflammatory action thereby protecting the animal from endotoxemia-induced multi-organ damage.

Effects of yoga training and detraining on physical performance measures in prepubertal children--a randomized trial.

Purpose of the study was to evaluate the effect of yoga training and detraining on physical performance measures in pre-pubertal (7-9 year old) school going children. Subjects were randomized to two groups - yoga group and Physical exercise (PE) group after the baseline assessment. All the subjects were assessed for strength, endurance, whole body endurance through 20 meter shuttle and physical fitness, at 3 time points - Baseline, 3 months Post intervention and 3 months after detraining. The results suggest that the improvement in the physical performance is largely by the increase in the respiratory muscle strength in the yoga group. In conclusion, the study presents the efficacy of yoga to improve strength, endurance, whole body endurance and aerobic capacity with 3 months of training in the pediatric group. However, the effect of the training does not last after 3 months detraining.

Effect of yoga training and detraining on respiratory muscle strength in pre-pubertal children: A randomized trial.

OBJECTIVE: To evaluate the effect of yoga on forced vital capacity (FVC), forced expiratory volume in I(st) second (FEV1), peak expiratory flow rate (PEFR), FEVI/FVC ratio, and pulmonary pressures [maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP) at the end of 3 months yoga training and the detraining effect on the above parameters in 7-9-years-old school going children. MATERIALS AND METHODS: A total of 100 participants were recruited from a school in Bangalore. After baseline assessments, the participants were randomly allocated to either yoga or physical activity group. Intervention was given for 3 months, and measures of pulmonary function and pulmonary pressures were determined immediately post-intervention and at 3-months follow-up. RESULTS: Although significant increase was observed in FVC, FEV1, PEFR, FEV1/FVC, MIP, and MEP at post-intervention, there were no significant differences between the two study groups after adjusting for height and age post training . However, MIP increased significantly in both the groups post-intervention, but the yoga group performed significantly higher than the PE group. The effects of training did not fade off even after 3 months of detraining. In fact, the FVC and FEV1 continued to increase significantly. A trend of decrease was observed in PEFR, MIP, and MEP. However, the values did not regress to the baseline value. CONCLUSIONS: This study suggests that practice of yoga for a short duration (3 months) of time can significantly improve respiratory muscle strength in pediatric population.

Effects of physical activity on pupil cycle time (PCT) in healthy Indian male.

Globally, physical inactivity is an important risk factor for the development of non-communicable disease consisting of coronary artery disease, as well as, other diseases including hypertension, diabetes, obesity, osteoporosis, and certain types of cancers. Parasympathetic nervous system (PNS) activity in the eye is determined by the pupil cycle time (PCT) can be comparable with cardiac parasympathetic response and thereby determine the morbidity and mortality among individuals. The PCT is measured by throwing white light on the edge of the pupil. Pupil cycling is a feature of pupillary reflex arc. The aim of this study is to establish the effect of physical activity on the PCT. The counting of PCT was done for 90 cycles and average one count is considered a single PCT. The physical activity level (PAL) was determined by administering a physical activity level questionnaire developed in the Division of Nutrition, St. John's Medical College, Bangalore. The PAL is classified as < 1.4 as sedentary, 1.55 to 1.75 moderately active, and > 1.75 heavily active. Thirty healthy male volunteers in the age group of 18-50 years and with BMI of 18.5 kg/m2-30 kg/m2 were studied. We obtained PCT of 962.00 ± 105.72 msec in sedentary, 896.77 ± 85.88 msec in moderately active and 889.45 ± 68.71 msec in heavily active individuals. Linear regression analysis shows there is statistically significant difference between the three different groups of physical activity level with a b value of 0 and R2 being 0.19. Increase in physical activity led to decrease in the PCT i.e. increase in the parasympathetic tone in the eye. Pupil cycle time (PCT) is a simple noninvasive tool to assess and differentiate the PNS function in different activity level of individual.

VO2 max in an Indian population: a study to understand the role of factors determining VO2 max.

VO2 max is the maximum amount of oxygen a person can consume and the value does not change despite an increase in workload. There is lack of data evaluating the impact of factors, which could affect VO2 max measurement, particularly in Indian population. The objectives of the present study were (i) to estimate VO2 max in a young healthy Indian population and to compare it with available prediction equations for Indian population (ii) to correlate time to achieve VO2 max with the relative VO2 max (iii) to assess the factors affecting the time to achieve VO2 max measurement (body composition and physical activity level). Twenty healthy adult males (18-30 years) underwent detailed anthropometry, physical activity level and modified Bruce protocol for VO2 max assessment. Breath by breath VO2, VCO2, oxygen saturation, heart rate, blood pressure were measured continuously and following exercise protocol. There was an internal validity between the estimated VO2 max and the maximum heart rate (MHR) (r = 0.51, P < 0.05). Respiratory rate and tidal volume significantly correlated with VO2 max P < 0.01). Linear regression analysis indicated physical activity level (PAL) was a strong predictor of time to reach VO2 max. Out of the 3 prediction equations computed to estimate VO2 max, 2 equations significantly overestimated VO2 max. In Conclusion, physical activity level emerged to be a strong predictor of time to VO2 max. Time to achieve VO2 max is an important factor to be considered when determining VO2 max. There was an overestimation in the VO2 max values derived from predicted equations.

Effect of short duration aerobic exercise training on reflection index, stiffness index and pulse wave velocity.

The study was aimed to establish the effect of aerobic exercise on reflection index (RI), stiffness index (SI) and brachial finger pulse wave velocity (BFPWV) in healthy subjects of Bangalore, India. Students of 18 to 25 years (males) were recruited according to the inclusion criteria. The subjects were trained on treadmill for a period of 8 weeks, and the above parameters were measured using BIOPAC software and doppler machine. ANOVA of repeated measures was done to see the effect of training on RI, SI and BFPWV with bonferroni correction. There was a significant decrease in above parameters (P<0.0001) compared to basal levels. There was significant correlation between BFPWV and SI (P<0.001). 8 weeks of training showed beneficial effect by reducing BFPWV, SI, and RI and it returned towards baseline after subsequent 4 weeks of detraining. We conclude that changes are because of the effect of aerobic exercise and are reversible.

Twist1 suppresses senescence programs and thereby accelerates and maintains mutant Kras-induced lung tumorigenesis.

KRAS mutant lung cancers are generally refractory to chemotherapy as well targeted agents. To date, the identification of drugs to therapeutically inhibit K-RAS have been unsuccessful, suggesting that other approaches are required. We demonstrate in both a novel transgenic mutant Kras lung cancer mouse model and in human lung tumors that the inhibition of Twist1 restores a senescence program inducing the loss of a neoplastic phenotype. The Twist1 gene encodes for a transcription factor that is essential during embryogenesis. Twist1 has been suggested to play an important role during tumor progression. However, there is no in vivo evidence that Twist1 plays a role in autochthonous tumorigenesis. Through two novel transgenic mouse models, we show that Twist1 cooperates with Kras(G12D) to markedly accelerate lung tumorigenesis by abrogating cellular senescence programs and promoting the progression from benign adenomas to adenocarcinomas. Moreover, the suppression of Twist1 to physiological levels is sufficient to cause Kras mutant lung tumors to undergo senescence and lose their neoplastic features. Finally, we analyzed more than 500 human tumors to demonstrate that TWIST1 is frequently overexpressed in primary human lung tumors. The suppression of TWIST1 in human lung cancer cells also induced cellular senescence. Hence, TWIST1 is a critical regulator of cellular senescence programs, and the suppression of TWIST1 in human tumors may be an effective example of pro-senescence therapy.

Bacopa monniera (L.) Wettst ameliorates behavioral alterations and oxidative markers in sodium valproate induced autism in rats.

Early prenatal or post natal exposure to environmental insults such as valproic acid (VPA), thalidomide and ethanol could induce behavioral alterations similar to autistic symptoms. Bacopa monniera, a renowned plant in ayurvedic medicine is useful in several neurological disorders. The purpose of the present study was to evaluate the effect of B. monniera on VPA induced autism. On 12.5 day of gestation the female pregnant rats were divided into control and VPA treated groups. They were administered saline/VPA (600 mg/kg, i.p.) respectively and allowed to raise their own litters. Group I-male pups of saline treated mothers. On postnatal day (PND) 21 VPA induced autistic male pups were divided into two groups (n = 6); Group II-received saline and Group III-received B. monniera (300 mg/kg/p.o.) from PND 21-35. Behavioral tests (nociception, locomotor activity, exploratory activity, anxiety and social behavior) were performed in both adolescence (PND 30-40) and adulthood (PND 90-110) period. At the end of behavioral testing animals were sacrificed, brain was isolated for biochemical estimations (serotonin, glutathione, catalase and nitric oxide) and histopathological examination. Induction of autism significantly affected normal behavior, increased oxidative stress and serotonin level, altered histoarchitecture of cerebellum (decreased number of purkinje cells, neuronal degeneration and chromatolysis) when compared with normal control group. Treatment with B. monniera significantly (p < 0.05) improved behavioral alterations, decreased oxidative stress markers and restored histoarchitecture of cerebellum. In conclusion, the present study suggests that B. monniera ameliorates the autistic symptoms possibly due to its anti-anxiety, antioxidant and neuro-protective activity.

Gender differences in age-related changes in cardiac autonomic nervous function.

Ageing is associated with changes in cardiac autonomic control as measured by Heart Rate Variability (HRV). Not many studies have explored the influence of gender on age-related changes in cardiac autonomic regulation. This study evaluated the gender differences in age-associated changes in cardiac autonomic nervous activity by assessing HRV using frequency domain analysis of short-term stationary R-R intervals. HRV was studied in healthy males and females ranging in age from 6 to 55 years. Total power and absolute power in High-Frequency (HF) and Low-Frequency (LF) components as well as HF in normalized unit declined significantly with ageing. The HF/LF ratio was significantly higher in the adolescent and adult females compared to male of these age groups. This study suggests that gender differences exist in age-related changes in HRV. The finding that gender differences are limited to adolescent and adult age groups may indicate a role for female sex hormones in cardiac autonomic modulation.

Probing the role of CXC motif in chemokine CXCL8 for high affinity binding and activation of CXCR1 and CXCR2 receptors.

All chemokines share a common structural scaffold that mediate a remarkable variety of functions from immune surveillance to organogenesis. Chemokines are classified as CXC or CC on the basis of conserved cysteines, and the two subclasses bind distinct sets of GPCR class of receptors and also have markedly different quaternary structures, suggesting that the CXC/CC motif plays a prominent role in both structure and function. For both classes, receptor activation involves interactions between chemokine N-loop and receptor N-domain residues (Site-I), and between chemokine N-terminal and receptor extracellular/transmembrane residues (Site-II). We engineered a CC variant (labeled as CC-CXCL8) of the chemokine CXCL8 by deleting residue X (CXC → CC), and found its structure is essentially similar to WT. In stark contrast, CC-CXCL8 bound poorly to its cognate receptors CXCR1 and CXCR2 (K(i) > 1 µm). Further, CC-CXCL8 failed to mobilize Ca(2+) in CXCR2-expressing HL-60 cells or recruit neutrophils in a mouse lung model. However, most interestingly, CC-CXCL8 mobilizes Ca(2+) in neutrophils and in CXCR1-expressing HL-60 cells. Compared with the WT, CC-CXCL8 binds CXCR1 N-domain with only ∼5-fold lower affinity indicating that the weak binding to intact CXCR1 must be due to its weak binding at Site-II. Nevertheless, this level of binding is sufficient for receptor activation indicating that affinity and activity are separable functions. We propose that the CXC motif functions as a conformational switch that couples Site-I and Site-II interactions for both receptors, and that this coupling is critical for high affinity binding but differentially regulates activation.

Comparison of cardiac autonomic activity between pre and post menopausal women using heart rate variability.

Ageing is associated with a decline in short-term indexes of heart rate variability (HRV). But there is little evidence regarding the extent to which age-related changes in HRV depend on simultaneous changes in levels of estrogen and body composition as it occurs from pre menopausal state to postmenopausal state. The purpose of this study was (i) to compare HRV between pre and postmenopausal women, (ii) to determine whether difference in age, estrogen level and body composition could account for the difference in HRV between these two groups. HRV was assessed using spectral analysis and estrogen level was estimated using radioimmunoassay technique. The body composition, in terms of percent fat, was assessed using measurement of skin fold thickness. Data was analyzed both before and after adjusting for age, estrogen level and body composition. It was found that the total power, high frequency (HF) and the low frequency (LF) power spectrum of HRV in absolute units were significantly lower (P < 0.001) in postmenopausal women compared to that of premenopausal women. Postmenopausal women had significantly lower HF (P < 0.01) and higher LF (P < 0.01) when expressed in normalized units. The ratio of LF/HF, the index of sympathovagal balance was significantly higher (P < 0.01) among postmenopausal women. Analysis after adjusting for age, revealed that age was one of the important confounder, responsible for the differences in all the components of power spectrum between the two study groups. Difference in estrogen level contributed for the difference in relative values of HF and LF components of HRV. Difference in body composition did not explain the difference in HRV between the groups. The study concludes that both ageing and declined estrogen levels are associated with the autonomic alterations seen among postmenopausal women.

Protection against radiation oxidative damage in mice by Triphala.

Protection against whole body gamma-irradiation (WBI) of Swiss mice orally fed with Triphala (TPL), an Ayurvedic formulation, in terms of mortality of irradiated animals as well as DNA damage at cellular level has been investigated. It was found that radiation induced mortality was reduced by 60% in mice fed with TPL (1g/kg body weight/day) orally for 7 days prior to WBI at 7.5 Gy followed by post-irradiation feeding for 7 days. An increase in xanthine oxidoreductase activity and decrease in superoxide dismutase activity was observed in the intestine of mice exposed to WBI, which, however, reverted back to those levels of sham-irradiated controls, when animals were fed with TPL for 7 days prior to irradiation. These data have suggested the prevention of oxidative damage caused by whole body radiation exposure after feeding of animals with TPL. To further understand the mechanisms involved, the magnitude of DNA damage was studied by single cell gel electrophoresis (SCGE) in blood leukocytes and splenocytes obtained from either control animals or those fed with TPL for 7 days followed by irradiation. Compared to irradiated animals without administering TPL, the mean tail length was reduced about three-fold in blood leukocytes of animals fed with TPL prior to irradiation. Although, similar protection was observed in splenocytes of TPL fed animals, the magnitude of prevention of DNA damage was significantly higher than that observed in leukocytes. It has been concluded that TPL protected whole body irradiated mice and TPL induced protection was mediated through inhibition of oxidative damage in cells and organs. TPL seems to have potential to develop into a novel herbal radio-protector for practical applications.

Potential of traditional ayurvedic formulation, Triphala, as a novel anticancer drug.

The cytotoxic effects of aqueous extract of Triphala, an ayurvedic formulation, were investigated on human breast cancer cell line (MCF-7) and a transplantable mouse thymic lymphoma (barcl-95). The viability of treated cells was found to decrease with the increasing concentrations of Triphala. On the other hand, treatment of normal breast epithelial cells, MCF-10 F, human peripheral blood mononuclear cells, mouse liver and spleen cells, with similar concentrations of Triphala did not affect their cytotoxicity significantly. The drug treatment was found to induce apoptosis in MCF-7 and barcl-95 cells in vitro as determined by annexin-V fluorescence and proportion of apoptotic cells was found dependent on Triphala concentration. MCF-7 cells treated with Triphala when subjected to single cell gel electrophoresis, revealed a pattern of DNA damage, characteristic of apoptosis. Studies on Triphala treated MCF-7 and barcl-95 cells showed significant increase in intracellular reactive oxygen species (ROS) in a concentration dependent manner. ROS increase was, however, found to be insignificant in MCF-10 F as well as in murine spleen and liver normal cells. In vivo, direct oral feeding of Triphala to mice (40 mg/kg body weight) transplanted with barcl-95 produced significant reduction in tumor growth as evaluated by tumor volume measurement. It was also found that apoptosis was significantly higher in the excised tumor tissue of Triphala fed mice as compared to the control, suggesting the involvement of apoptosis in tumor growth reduction. These results suggest that Triphala possessed ability to induce cytotoxicity in tumor cells but spared the normal cells. The differential effect of Triphala on normal and tumor cells seems to be related to its ability to evoke differential response in intracellular ROS generation. The differential response of normal and tumor cells to Triphala in vitro and the substantial regression of transplanted tumor in mice fed with Triphala points to its potential use as an anticancer drug for clinical treatment.

Cytotoxic response of breast cancer cell lines, MCF 7 and T 47 D to triphala and its modification by antioxidants.

The cytotoxic effects of Triphala (TPL), an Indian Ayurvedic formulation with known anti-cancer properties, has been investigated on two human breast cancer cell lines differing in their p53 status. In vitro studies showed that MCF 7 with wild type p53 was more sensitive to TPL than T 47 D, which is p53 negative. TPL induced loss of cell viability was determined by MTT assay. After 72h incubation, the IC 50 values for MCF 7 was found to be approximately 8microg/ml and that for T 47 D was approximately 26microg/ml. Moreover, TPL inhibited the clonogenic growth of MCF 7 cells, which was significantly recovered by pifithrin-alpha, the p53 inhibitor. However, pifithrin-alpha, did not modify TPL induced cytotoxicity in T 47 D cells. Exogenous addition of antioxidants, glutathione (GSH) and N-Acetyl-Cysteine (NAC) inhibited the anti-proliferative ability of TPL in both MCF 7 and T47 D. Annexin-V and propidium iodide double staining of cells treated with TPL for 2h revealed that TPL induced significant apoptosis in both the cell lines in a dose dependant manner but magnitude of apoptosis was significantly higher in MCF 7 than in T 47-D cells. TPL was also found to induce dose and time dependent increase in intracellular reactive oxygen species in both the cell lines. Present results have demonstrated that MCF 7 and T 47 D cells exhibited differential sensitivity to TPL, which seems to be dependant on their p53 status. Inhibition of anti-proliferative ability of TPL by antioxidants suggests a role for TPL induced ROS in the induction of apoptosis. It is concluded that p53 status of cancer cells formed an important factor in predicting the response of cancer cells to prooxidant drugs.