Association of Stressful Life Events with Dementia in North Indian Populations.

BACKGROUND: Chronic stress is known to be associated with dementia. AIM: This study looked for major stressors and their associations with dementia in the elderly demented population. METHODS: Case group consisted of 60 subjects (age group 55-75 years; M:F = 7:3) with postscreening for mild cognitive impairment (MCI) and mild Alzheimer's type dementia. Control group consisted of two subgroups: related (N = 60) and not-related (N = 60) subjects screened and confirmed not to be suffering from cognitive impairment. Sociodemographic variables were collected using a personal data sheet. The severity of cognitive impairment was assessed using the Clinical Dementia Rating (CDR) Scale. Subjects with a CDR score of range 0.5-1 were included in mild cognitive impairment and mild Alzheimer's type dementia. Stressful life events were evaluated using the Presumptive Stressful Life Event Scale (PSLES). A present stress level of caregivers was evaluated using Depression Anxiety Stress Scales (DASS). Assessed stress level scores were compared and validated with serum and plasma levels of cortisol and ACTH respectively. RESULTS: Different groups exhibited distinct PSLES and DASS scores with statistical significance. It was also observed that Cortisol and ACTH levels were elevated in case group as compared with that of both sub-group controls. CONCLUSIONS: The outcome measures of this study projected that neural degeneration leading to mild cognitive impairment and dementia may be due to accumulative cortisol hormone associated with stressful life events.

Accelerated identification of serine racemase inhibitor from Centella asiatica.

Serine racemase (SR) converts the free form of L-serine into D-serine (DS) in the mammalian brain. The DS functions as a co-agonist of N-methyl D-aspartate (NMDA) receptor. The over- activation of NMDA receptor leads to many neurological disorders like stroke, amyotrophic lateral sclerosis, Alzheimer's disease and an effective inhibitor of SR could be a corrective method for the receptor over-activation. We report for the first time here a rapid way of purifying and identifying an inhibitor from medicinal plants known to have the neuro-protective effect. We have purified SR inhibitor from the methanolic extract of Centella asiatica by affinity method. High resolution mass spectrometry and infrared spectroscopy were used to identify the ligand to be madecassoside. We have shown the madecassoside binding in silico and its inhibition of recombinant human serine racemase in vitro and ex vivo.

In vivo automated quantification of thermally damaged human tissue using polarization sensitive optical coherence tomography.

Over the decades numerous technologies have been performed for the quantification of skin injuries, but their poor sensitivity, specificity and accuracy limits their applications. Optical coherence tomography (OCT) can be potential tool for the identification but the modern high-speed OCT system acquires huge amount of data, which will be very time-consuming and tedious process for human interpretation. Our proposed method opens the possibility of fully automated quantitative analysis based on morphological features of thermally damaged tissue, which will become biomarker for the removal of non-viable skin. The proposed method is based on multi-level ensemble classifier by dissociating morphological features (A-line, B-scan, phase images) extracted from Polarization Sensitive Optical Coherence Tomography (PS-OCT) images. Our proposed classifier attained the average sensitivity, specificity and accuracy is 92.22%, 87.2% and 92.5%, respectively, in detecting the thermally damaged human skin. Moreover, we show that our classifier is one of the best possible classifier based on features extracted from PS-OCT images, which demonstrates the significance of PS-OCT data in detecting abnormality in human skin.

Estimation of Serum YKL-40 by Real-Time Surface Plasmon Resonance Technology in North-Indian Asthma Patients.

BACKGROUND: Many studies reported for estimating serum YKL-40 using ELISA or RIA methods. This study introduces the plausible utilization of real-time surface plasmon resonance (SPR) technology in investigating the expression of serum YKL-40 protein levels and ELISA method for serum IgE in bronchial asthma. METHODS: A commercially available BIAcore 2000 instrument, based on SPR technology, was utilized for assessing serum YKL-40 levels in a control sample size of 45 and active sample size of 97. Antibody immobilization was optimized to obtain the best sensor performance and a sensitive analytic detection. A commercially available ELISA kit was utilized for detecting serum IgE to estimate allergic condition-associated asthma. RESULTS: The results of SPR technology could distinctly classify with highly statistical significance, the asthma severities by estimating the elevated levels of YKL-40 in blood sera of minute quantities (up to 0.33 ng/ml), and thus differentiates superior utility in comparison with ELISA method. No statistically significant correlation of YKL-40 and IgE was observed. CONCLUSIONS: Serum YKL-40 may be used as a protein marker in classifying asthma severity by applying SPR technology as a reliable, label-free, highly sensitive, and cost-effective tool.

Pharmacophore development and screening for discovery of potential inhibitors of ADAMTS-4 for osteoarthritis therapy.

In the development of osteoarthritis, aggrecan degrades prior to cartilage destruction. Aggrecanase-1 (ADAMTS-4) is considered to be the major enzyme responsible for cleaving the Glu373-Ala374 bond in the interglobular domain of aggrecan in humans. Therefore, inhibitors of ADAMTS-4 have therapeutic potential in the treatment of osteoarthritis. In the present work, we developed a chemical feature based pharmacophore model of ADAMTS-4 inhibitors using the HipHop module within the Catalyst program package in order to elucidate the structure-activity relationship and to carry out in-silico screening. The Maybridge database was screened using Hypo1 as a 3D query, and the best-fit hits that followed Lipinski's rule of five were subsequently screened to select the compounds. The hit compounds were then docked into the active site of ADAMTS-4, and interactions were visualized to determine the potential lead molecules. After subjecting all of the hits to various screening and filtering processes, 13 compounds were finally evaluated for their in vitro inhibitory activities. This study resulted in the identification of two lead compounds with potent inhibitory effects on ADAMTS-4 activity, with IC50 values of 0.042 µM and 0.028 µM, respectively. These results provide insight into the pharmacophoric requirements for the development of more potent ADAMTS-4 inhibitors. Graphical Abstract The aggrecan-degrading metalloprotease ADAMTS-4 has been identified as a novel therapeutic target for osteoarthritis. In this work, we used HipHop-based pharmacophore modeling and virtual screening of the Maybridge database to identify novel ADAMTS-4 inhibitors. These novel lead compounds act as potent and specific inhibitors for the ADAMTS-4 enzyme and could have therapeutic potential in the treatment of OA.

Association of polymorphisms in angiotensin-converting enzyme gene with gestational diabetes mellitus in Indian women.

BACKGROUND: Numerous genes have been reported in relation with gestational diabetes mellitus (GDM), but the findings were not consistently replicated across populations, or there have been no detailed studies on them. Previous literatures suggested that, out of all angiotensin converting enzyme (ACE) gene polymorphisms, only ACE insertion/deletion (I/D) gene polymorphism has a strong association with GDM in Asian Indian women. AIM: This study was devoted to evaluate the association of four single nucleotide polymorphisms (SNPs) ACE A240T, C1237T, G2350A and I/D with GDM and Type 2 diabetes mellitus. MATERIALS AND METHODS: This study recruited 105 GDM cases, 119 Type 2 diabetes mellitus subjects and 120 controls. PCR-RFLP was used for identifying genotypes of ACE A240T, C1237T and G2350A and PCR was performed in the case of ACE I/D. RESULTS: Significant associations of ACE SNP's, C1237T, and G2350A with GDM were observed. Haplotype analysis revealed the remarkably significant evidence of association with SNP combination ACE A240T, C1237T, G2350A, and I/D with GDM patients (P = 0.024). Individuals possessing haplotype "TTAI" (frequency 30% in GDM and 0 in controls) derived from these SNPs had 185 fold increased risk of developing GDM (95% of confidence interval: 11.13-3102.15), which was highest when compared with other 15 haplotypes. CONCLUSION: Shorter-range haplotypes were also significant, but the only consistently associated alleles were found to be in ACE C1237T, G2350A, and I/D. These results suggested that the variant in close proximity to ACE C1237T, G2350A and/or I/D modulates susceptibility to GDM and noninsulin dependent diabetes mellitus in Indian women.

Determination of efficacy of reflexology in managing patients with diabetic neuropathy: a randomized controlled clinical trial.

Background. The restricted usage of existing pharmacological methods which do not seem to provide the treatment of diabetic neuropathy may lead to exploring the efficacy of a complementary therapy. In this context, this paper was devoted to evaluate the efficacy of foot reflexology. This health science works on the hypothesis that the dysfunctional states of body parts could be identified by observing certain skin features and be rectified by stimulating certain specific areas mapped on feet. Method. Subjects (N = 58) with diagnosed diabetic neuropathy were randomly distributed into reflexology and control groups in which both group patients were treated with ongoing pharmacological drugs. Reflexology group patients were additionally treated holistically with the hypothesis that this therapy would bring homeostasis among body organ functions. This was a caregiver-based study with a follow-up period of 6 months. The outcome measures were pain reduction, glycemic control, nerve conductivity, and thermal and vibration sensitivities. The skin features leading to the detection of the abnormal functional states of body parts were also recorded and analyzed. Results. Reflexology group showed more improvements in all outcome measures than those of control subjects with statistical significance. Conclusion. This study exhibited the efficient utility of reflexology therapy integrated with conventional medicines in managing diabetic neuropathy.

Serum based diagnosis of asthma using Raman spectroscopy: an early phase pilot study.

The currently prescribed tests for asthma diagnosis require compulsory patient compliance, and are usually not sensitive to mild asthma. Development of an objective test using minimally invasive samples for diagnosing and monitoring of the response of asthma may help better management of the disease. Raman spectroscopy (RS) has previously shown potential in several biomedical applications, including pharmacology and forensics. In this study, we have explored the feasibility of detecting asthma and determining treatment response in asthma patients, through RS of serum. Serum samples from 44 asthma subjects of different grades (mild, moderate, treated severe and untreated severe) and from 15 reference subjects were subjected to Raman spectroscopic analysis and YKL-40 measurements. The force expiratory volume in 1 second (FEV1) values were used as gold standard and the serum YKL-40 levels were used as an additional parameter for diagnosing the different grades of asthma. For spectral acquisition, serum was placed on a calcium fluoride (CaF2) window and spectra were recorded using Raman microprobe. Mean and difference spectra comparisons indicated significant differences between asthma and reference spectra. Differences like changes in protein structure, increase in DNA specific bands and increased glycosaminoglycans-like features were more prominent with increase in asthma severity. Multivariate tools using Principal-component-analysis (PCA) and Principal-component based-linear-discriminant analysis (PC-LDA) followed by Leave-one-out-cross-validation (LOOCV), were employed for data analyses. PCA and PC-LDA results indicate separation of all asthma groups from the reference group, with minor overlap (19.4%) between reference and mild groups. No overlap was observed between the treated severe and untreated severe groups, indicating that patient response to treatment could be determined. Overall promising results were obtained, and a large scale validation study on random subjects is warranted before the routine clinical usage of this technique.

Role of reflexology and antiepileptic drugs in managing intractable epilepsy--a randomized controlled trial.

BACKGROUND: This report is based on the results of a randomized parallel controlled trial conducted to determine the efficacy of reflexology therapy in managing intractable epilepsy. METHODS: Subjects who failed epilepsy surgery or were not candidates for epilepsy surgery or were non-responders of antiepileptic drugs (AEDs) took part in this study. The trial was completed by 77 subjects randomly assigned to 2 arms: control (AEDs) and reflexology (AEDs + reflexology therapy). The hypothesis was that hand reflexology therapy could produce results similar to those of vagus nerve stimulation, and foot reflexology therapy could maintain homeostasis in the functional status of individual body parts. Reflexology therapy was applied by family members. The follow-up period was 1.5 years. Quality of life in epilepsy patients was assessed with the QOLIE-31 instrument. RESULTS: In the reflexology group, the median baseline seizure frequency decreased from 9.5 (range 2-120) to 2 (range 0-110) with statistical significance (p < 0.001). In the control arm, the decrease was less than 25% with a baseline value of 16 (range 2-150). The pretherapy QOLIE-31 scores in the control group and the reflexology group were 41.05 ± 7 and 43.6 ± 8, respectively. Posttherapy data were 49.07 ± 6 and 65.4 ± 9, respectively (p < 0.002). The reflexology method allowed detection of knee pain in 85% of the reflexology group patients (p < 0.001), and 85.3% of patients derived 81% relief from it (p < 0.001). 4 reflexology group patients reported nausea/vomiting (n = 1), change in voice (n = 2), and hoarseness (n = 1). CONCLUSION: Reflexology therapy together with AEDs may help reducing seizure frequency and improving quality of life in individuals with epilepsy.

Noninvasive characterisation of foot reflexology areas by swept source-optical coherence tomography in patients with low back pain.

Objective. When exploring the scientific basis of reflexology techniques, elucidation of the surface and subsurface features of reflexology areas (RAs) is crucial. In this study, the subcutaneous features of RAs related to the lumbar vertebrae were evaluated by swept source-optical coherence tomography (SS-OCT) in subjects with and without low back pain (LBP). Methods. Volunteers without LBP (n = 6 (male : female = 1 : 1)) and subjects with LBP (n = 15 (male : female = 2 : 3)) were clinically examined in terms of skin colour (visual perception), localised tenderness (visual analogue scale) and structural as well as optical attributes as per SS-OCT. From each subject, 6 optical tomograms were recorded from equidistant transverse planes along the longitudinal axis of the RAs, and from each tomogram, 25 different spatial locations were considered for recording SS-OCT image attributes. The images were analysed with respect to the optical intensity distributions and thicknesses of different skin layers by using AxioVision Rel. 4.8.2 software. The SS-OCT images could be categorised into 4 pathological grades (i.e., 0, 1, 2, and 3) according to distinctness in the visible skin layers. Results. Three specific grades for abnormalities in SS-OCT images were identified considering gradual loss of distinctness and increase in luminosity of skin layers. Almost 90.05% subjects were of mixed type having predominance in certain grades. Conclusion. The skin SS-OCT system demonstrated a definite association of the surface features of healthy/unhealthy RAs with cutaneous features and the clinical status of the lumbar vertebrae.

Serum cartilage oligomeric matrix protein (COMP) in knee osteoarthritis: a novel diagnostic and prognostic biomarker.

A case-control study was conducted to estimate the association of cartilage oligomeric matrix protein (COMP) with knee osteoarthritis (OA) and to examine the potential utility of COMP as a diagnostic and prognostic biomarker in early knee OA. The COMP levels were estimated in the blood sera of 150 subjects belonging to study group (n = 100) and control one (n = 50). Patients with confirmed clinical isolated knee OA diagnosed through American College of Rheumatology criteria were included and were without any other cause of knee pain. ELISA was used to determine the levels of COMP, interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α). The median (range) serum COMP levels were observed to be 1117.21 ng/ml (125.03-4209.75 ng/ml) in OA patients and 338.62 ng/ml (118-589 ng/ml) in control subjects with p < 0.001. The COMP levels of study group were negatively correlated (correlation factor -0.88) with disease duration and positively correlated with age, BMI, pain score and IL-1ß with correlation factors 0.86, 0.63, 0.76, and 0.79, respectively with p < 0.001. Gender differentiation was found in study group with 52% higher COMP level in males as compared to that of females. There was no significant correlation of COMP levels with radiological grading, erythrocyte sedimentation rate (ESR), hemoglobin (Hb), and TNF-α. The serum COMP levels may be used as a diagnostic OA marker along with prognostic value in determining the patients at risk of rapidly progressing this debilitating joint disease. The serum COMP level remains significantly high in first 3 years of disease duration.

Optical, spectroscopic, and Doppler evaluation of "normal" and "abnormal" reflexology areas in lumbar vertebral pathology: a case study.

Scientific validation of reflexology requires an in-depth and noninvasive evaluation of "reflexology/reflex areas" in health and disease. The present paper reports the differential properties of "normal" and "abnormal" reflexology areas related to the lumbar vertebrae in a subject suffering from low back pain. The pathology is supported by radiological evidence. The reflexology target regions were clinically assessed with respect to colour and tenderness in response to finger pressure. Grey scale luminosity and pain intensity, as assessed by visual analogue scale scores, differentiated "normal" from "abnormal" skin. Skin swept source-optical coherence tomography recorded their structural differences. Infrared thermography revealed temperature variations. A laser Doppler study using a combined microcirculation and transcutaneous oxygen monitoring system indicated alterations in blood flow and oxygen perfusion. Raman spectroscopy showed differences in chemical signatures between these areas. The present findings may indicate a potential correlation between the reflexology areas and subsurface pathological changes, showing an association with the healthy or unhealthy status of the lumbar vertebrae.

ADAMTS-4 and ADAMTS-5: key enzymes in osteoarthritis.

Osteoarthritis (OA) is a progressive disease of the joints characterized by degradation of articular cartilage. Although disease initiation may be multi-factorial, the cartilage destruction appears to be a result of uncontrolled proteolytic extracellular matrix destruction. A major component of the cartilage extracellular matrix is aggrecan, a proteoglycan that imparts compressive resistance to the tissue. Aggrecanase-mediated aggrecan degradation is a significant event in early stage OA. The relative contribution of individual ADAMTS-4 and ADAMTS-5 proteinases to cartilage destruction during OA has not been resolved completely. This review reveals that both ADAMTS-4/ADAMTS-5 are responsible for aggrecan degradation in a human model of OA, and is expected to list down the rational strategies which are being focussed for therapeutic intervention in OA.

Prognostic significance of cyclooxygenase-2 and response to chemotherapy in invasive ductal breast carcinoma patients by real time surface plasmon resonance analysis.

Cyclooxygenase-2 (COX-2), an inducible enzyme, has been implicated in the progression and angiogenesis of breast cancer. The aim of the study is to quantify the concentration of COX-2 and its association with clinico-pathological parameters and response to treatment in patients with invasive ductal carcinoma receiving both neo-adjuvant and adjuvant chemotherapy. The level of COX-2 was estimated using a novel biosensor-based surface plasmon resonance technique in serum of 84 patients with breast cancer (48 patients of neo-adjuvant chemotherapy and 36 patients of adjuvant chemotherapy) and 40 age- and gender-matched normal individuals. A significant increase in COX-2 level was observed in patients compared with normal individuals (p>0.0001). The COX-2 level in serum was found to be significantly higher in patients with lymph node involvement (p<0.0061). 68% (33/48) of the patients receiving neo-adjuvant chemotherapy showed significantly (p<0.0025) reduced COX-2 levels. This study shows significant decrease of COX-2 level in patients with breast cancer treated with both neo-adjuvant and adjuvant chemotherapy. Estimation of COX-2 level in serum may serve as a tumor biomarker in patients with breast cancer.

Isolation and characterization of novel protein with anti-fungal and anti-inflammatory properties from Aloe vera leaf gel.

The Aloe protein of 14 kDa from the Aloe vera leaf gel was isolated by an ion exchange chromatography using DEAE-cellulose and CM-cellulose column. The purified Aloe protein exhibited a potent anti-fungal activity against Candida paraprilosis, Candida krusei and Candida albicans. In addition, the purified Aloe protein also showed an anti-inflammatory property against pure lipoxygenase and cyclooxygenase-2 with 84% and 73% inhibition, respectively, and was verified by binding with these proteins by real time method by the phenomenon of surface plasmon resonance. This Aloe protein is a novel protein possessing antifungal and anti-inflammatory properties and thus sets a platform to be used as a medicinal plant product.