Genetic diversity and population structure of critically endangered Dactylorhiza hatagirea (D. Don) Soo from North-Western Himalayas and implications for conservation.

Dactylorhiza hatagirea (D. Don) Soo is medicinally important herb, which is widely used in ayurveda, unani, and folk/traditional medicine system to cure diseases. Due to its immense ethno-botanical properties, the trade of D. hatagirea is estimated to be USD 1 billion/year in India. Unfortunately, due to overexploitation of the herb from the wild, has resulted in dwindling of its populations in their natural habitats, which has led to its critically endangered status. Molecular genetic studies are still scarce in D. hatagirea, therefore, in current study, genetic diversity and population structure analysis was carried out of 10 populations (48 individuals) collected from three cold desert regions (2527 m-3533 m amsl) of Himachal Pradesh. Mean observed heterozygosity (Ho) and expected heterozygosity (He) was recorded 0.185 and 0.158. The maximum values for Fst (fixation index) and Nm (gene flow) were recorded 0.945 at locus KSSR14 and 1.547 at locus KSSR 4 respectively. Mean genetic differentiation (Fst) coefficient was estimated to 0.542. Overall, low levels of genetic diversity was recorded in the populations of D. hatagirea, might be due to habitat specificity (alpine meadows ecosystem; humid laden undulating habitat), restricted distribution and high anthropogenic activities. However, two populations viz., Bathad and Rangrik were recorded with high diversity and largest number of private alleles, stipulates that these populations might have high evolutionary significance and response to selection. Dendrogram analysis revealed that the populations of D. hatagirea were clustered into four major clusters, which was supported by Bayesian based STRUCTURE predictions. Clustering pattern of majority individuals of different populations revealed consistency with their geographic origin. Outcomes of current study reveals the status of genetic diversity and population structure of endangered D. hatagirea, which can be futuristically utilised for appropriate planning of conservation strategies.

Management of Lateral Epicondylitis: A Prospective Comparative Study Comparing the Local Infiltrations of Leucocyte Enriched Platelet-Rich Plasma (L-aPRP), Glucocorticoid and Normal Saline.

Introduction: Lateral epicondylitis is a painful condition of the elbow, characterised by pain and tenderness with resisted wrist extension. This study was carried out to evaluate the comparative efficacy of the local infiltration of L-PRP, methylprednisolone and normal saline in patients with lateral epicondylitis. Materials and methods: Sixty adult patients, between the ages 30 to 50 years, diagnosed with lateral epicondylitis of more than 12 weeks, were enrolled in the prospective randomised study. Their medical history and previous conservative treatment were recorded; the clinical evaluation of the tendinitis was made with the visual analogue scale (VAS), the disabilities of the arm, shoulder, and hand (DASH) outcome scores, the modified elbow performance index (MEPS), the functional assessment by patient-rated tennis elbow evaluation (PRTEE), together with the laboratory investigations. The patients were randomised using the computer-generated alphabets into three groups of 20: group A received saline, group B received PRP, and group C received corticosteroids. Results: Patients were seen at 4, 8 and 12 weeks to evaluate the post-injection status. VAS, DASH, and PRTEE scores were significantly reduced, and MEPS was significantly improved in group B compared to group A and group C. Moreover, the reductions in VAS and PRTEE were significantly different in group C compared to group A. Conclusion: PRP leads to superior healing with long-term therapeutic advantages compared to corticosteroids though it takes a little longer to have its effect.

Myosteatosis, the More Significant Predictor of Outcome: An Analysis of the Impact of Myosteatosis, Sarcopenia, and Sarcopenic Obesity on Liver Transplant Outcomes in Johannesburg, South Africa.

OBJECTIVES: In high-income countries, myosteatosis, sarcopenia, and obesity with sarcopenia (sarcopenic obesity) are associated with adverse outcomes after liver transplantation. In South Africa, an upper-middleincome country, we investigated the prevalence and impact of these muscle abnormalities on posttransplant outcomes in adult liver transplant recipients. MATERIALS AND METHODS: We reviewed 106 liver transplant recipients and measured muscle abnormalities on computed tomography using segmentation software. The parameters evaluated were myosteatosis by mean muscle attenuation, sarcopenia by skeletal muscle index at the third lumbar vertebra using validated cutoffs, and sarcopenic obesity as sarcopenia and a body mass index of ≥25 kg/m². The effects of these abnormalities on 1-year patient and graft survival (primary endpoint) and length of hospital and intensive care unit stay, costs, and 90-day and overall postoperative complications (secondary endpoints) were assessed. RESULTS: Most liver transplant recipients were male (n = 64, 60%). Alcoholic and/or nonalcoholic steatohepatitis were the most frequent indications for transplant (n = 38, 36%). Myosteatosis occurred in 76 patients (72%), 69 patients (65%) had sarcopenia, and 36 patients (34%) had sarcopenic obesity. One year after transplant, myosteatosis was associated with higher mortality (hazard ratio of 3.3; 95% confidence interval, 1.00-11.13; P = .049), greater risk of allograft failure (hazard ratio of 4.1; 95% confidence interval, 1.2-13.5; P = .021), and longer hospital and intensive care unit stays compared with those without myosteatosis. All patients with no body composition abnormalities were alive at 1 year compared with 69% with coexisting myosteatosis and sarcopenia. CONCLUSIONS: In our setting, liver transplant recipients with myosteatosis had a higher risk of death and allograft failure at 1 year compared with patients without body composition abnormalities.

Lactobacillus fermentum (MTCC-5898) alleviates Escherichia coli-induced inflammatory responses in intestinal epithelial cells by modulating immune genes and NF-κB signalling.

AIM: Dietary intervention using probiotic bacteria has emerged as a promising preventive strategy in addressing foodborne infections or gastrointestinal disorders. This study investigated the immunomodulatory effects of Lactobacillus fermentum (MTCC-5898) on Escherichia coli-induced inflammatory responses in intestinal epithelial cells. METHODS AND RESULTS: The immune response of intestinal cells (Caco-2) in the presence of probiotic Lact. fermentum was determined during exclusion, competition and displacement of E. coli as the inflammatory agent. To achieve this objective, transcriptional modulation of key immune-related genes (cytokines, pattern recognition receptors and NF-κB), release of cytokines and nuclear translocation of the NF-κB subunit p-65 were studied. Expression of pro-inflammatory cytokines IL-8, TNF-α, IFN-ϒ and IL-23 was high in E. coli-exposed intestinal cells. However, overexpression of these E. coli-induced pro-inflammatory cytokines was prevented by Lact. fermentum during exclusion and competition assays. It also modulated the transcriptional expression of regulatory cytokines (IL-10 and TGF-ß), pattern recognition receptors (TLR-2 and TLR-4) and genes associated with master inflammatory regulators (NF-κB and SIGIRR) to reduce E. coli-induced inflammation. The protective effect of Lact. fermentum was further confirmed by suppression of nuclear translocation of cytoplasmic NF-κB subunit (p-65). CONCLUSION: Lactobacillus fermentum alleviated E. coli-induced inflammatory responses by modulating the NF-κB signalling besides pro-inflammatory and regulatory cytokines expression. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus fermentum holds significant promise as a potent nutraceutical that prevents and manages inflammatory gut-associated dysfunctions.

Protective effects of casein-derived peptide VLPVPQK against hydrogen peroxide-induced dysfunction and cellular oxidative damage in rat osteoblastic cells.

Oxidative stress inhibits osteoblast differentiation and function that lead to the development of osteoporosis. Casein-derived peptide VLPVPQK (PEP), a potent antioxidant, was isolated from ß-casein of buffalo milk. We used an in vitro oxidative stress model induced by hydrogen peroxide (H2O2) in rat osteoblastic cells to investigate the protective effects of PEP against H2O2-induced dysfunction and oxidative damage. Cells were pretreated with PEP (50-200 ng/mL) for 2, 7 or 21 days followed by 0.3 mM H2O2 treatment for 24 h and then markers of osteogenic development, oxidative damage and apoptosis were examined. PEP significantly increased the viability and differentiation markers of osteoblast cells such as alkaline phosphatase and calcium mineralization. Moreover, PEP suppressed the production of reactive oxygen species (ROS), lipid peroxidation and ameliorated H2O2-induced reduction in glutathione, superoxide dismutase and catalase activities. In addition, PEP partially inhibited caspase-9 and-3 activities and reduced propidium iodide-positive cells. Altogether, our results demonstrated that PEP could protect rat osteoblast against H2O2-induced dysfunction and oxidative damage by reduction of ROS production, lipid peroxidation and increased antioxidant enzyme activities. Thus, our data suggest that PEP might be a valuable protective agent against oxidative stress-related diseases such as osteoporosis.

Cross-talk between probiotic lactobacilli and host immune system.

The mechanism by which probiotic lactobacilli affect the immune system is strain specific. As the immune system is a multicompartmental system, each strain has its way to interact with it and induce a visible and quantifiable effect. This review summarizes the interplay existing between the host immune system and probiotic lactobacilli, that is, with emphasis on lactobacilli as a prototype probiotic genus. Several aspects including the bacterial-host cross-talk with the mucosal and systemic immune system are presented, as well as short sections on the competing effect towards pathogenic bacteria and their uses as delivery vehicle for antigens.

Evaluation of juxta-apical radiolucency in cone beam CT images.

OBJECTIVES: The aim of this study was to analyse the position and relationship of juxta-apical radiolucency (JAR) to the mandibular canal and buccal and/or lingual cortical plates using cone beam CT (CBCT). METHODS: A retrospective study was carried out to analyse the JAR on CBCT for 27 patients. These findings were compared with 27 age- and sex-matched patients without the presence of JAR, which acted as the control group. The CBCT images were analysed according to a checklist, to evaluate the position of the JAR and its relationship to the mandibular canal. Then, any thinning or perforation of either the buccal or lingual cortical plate due to JAR was noted, and a classification to quantify the thinning of cortical plates was proposed. The findings in the two groups were analysed using a paired comparison by McNemar test. RESULTS: A statistical increased thinning of cortical plates was seen in the JAR group compared with the control group, and most of the cases were in the J3 group. None of the patients in either the JAR or the control group showed perforation of the buccal and/or lingual cortical plate on CBCT images. CONCLUSIONS: A classification to quantify the thinning of cortical plates was proposed, which may be used for objective evaluation of the thinning of the cortical plates in future studies. The present study gives an insight into the relationship of the juxta-apical area with the mandibular canal and cortical plates in the mandible using CBCT.

Induction of immune tolerance to caseins and whey proteins by oral intubation in mouse allergy model.

This study was designed to evaluate the effect of oral tolerance of caseins (CSN) and whey proteins (WP) in alleviating the allergic response to cow's milk proteins in Swiss albino mice raised on a milk protein-free diet. Oral tolerance was induced by feeding mice with 20 mg of CSN or WP once in a day for 4 days consecutively before immunization with respective protein by intraperitoneal (i.p.) injections (20 µg 200 per µl of PBS) using 2% of alum Al(OH)3 as adjuvant. Three weeks later, oral tolerance induction was analysed in humoral and cellular compartments of CSN- and WP-fed versus saline-fed control mice groups by measuring seric and intestinal antibody responses, mRNA abundance in splenic tissue and cytokine secretion patterns. The specific serum immunoglobulin-E (IgE) levels were significantly suppressed (p < 0.05), while sIgA was enhanced in these groups when compared with their respective saline-fed mice. Moreover, the mRNA levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4) in both CSN- and WP-tolerized mice were found to be significantly decreased, while the abundance of interleukin-10 (IL-10) and transforming growth factor-ß (TGF-ß) was increased significantly, as compared to respective control groups. Finally, cytokine profiles indicated a reciprocal decrease in IL-4 and IFN-γ versus an increase in IL-10 secretions in supernatants of cultured splenocytes of tolerized mice. Taken together, these results clearly showed that oral administration of cows' milk caseins and whey proteins can induce significant hyposensitization in mice, with the participation of suppressor cytokines.

Effect of supplementation of vitamin E, copper and zinc on the in vitro phagocytic activity and lymphocyte proliferation index of peripartum Sahiwal (Bos indicus) cows.

To study the effect of vitamin E (VE), copper (Cu) and zinc (Zn) supplementation on the in vitro phagocytic activity (PA) and lymphocyte proliferation response (LPR) of blood neutrophils and lymphocytes, thirty Sahiwal pregnant cows (six in each group) in their late gestation at 30 days before the expected date of calving were selected from the NDRI experimental herd and supplemented with various micronutrients from 30 days before calving to 45 days after calving. Cows were supplemented individually with VE (1000 IU/cow/day), Cu (20 ppm/cow/day) and Zn (80 ppm/cow/day) and also with a combination of VE, Cu and Zn to study cumulative effect of all micronutrients. One group without any supplementation acted as a control. Blood neutrophils and lymphocytes were isolated and studied for their PA and LPR. Supplementation of micronutrients like VE, Cu, Zn and a combination of all these nutrients significantly (p < 0.01) increased the PA of experimental cows as compared to control (unsupplemented) cows during the pre-partum period. During post-partum, all the micronutrients (VE, Cu, Zn and their combination) showed a significant (p < 0.01) increase in the PA of experimental cows as compared to control cows. Of all the groups, significant (p < 0.01) and maximum PA was observed in the combination group followed by Zn-supplemented group during both the pre- and post-partum period. A significant (p < 0.01) increase in LPR of B lymphocytes was observed in combination-supplemented group during the pre-partum period and during both the pre- and post-partum period in the Cu-supplemented group.

In vitro phagocytic activity of milk neutrophils during lactation cycle in Murrah buffaloes of different parity.

Milk samples were collected from 34 lactating Murrah buffaloes on days 0, 5, 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, 270 and day 300 after calving. Milk somatic cell counts (SCC) were highest in multiparous buffaloes. Milk SCC were significantly lower in buffaloes of third and fourth parity during early lactation and than increased significantly (p < 0.01) by the end of lactation. Milk neutrophils were significantly lower in all the buffaloes during early lactation, but increased significantly (p < 0.01) afterwards. Milk lymphocytes were significantly higher during early lactation, but decreased significantly (p < 0.01) by the end of lactation. Phagocytic activity (PA) was highest in day 1 colostrum and then decreased significantly (p < 0.01) by the fourth milking in buffaloes of second, third and fourth parity. Phagocytic index (PI) was also highest in colostrums of primiparous buffaloes. Irrespective of parity, maximum PA and PI was observed during mid lactation. In terms of in vitro phagocytic activity, early lactation is the most critical period followed by late and mid lactation.

Skin conductance as a means to predict hypotension following spinal anaesthesia.

BACKGROUND AND OBJECTIVE: Hypotension following spinal anaesthesia (SA) is common, especially in the elderly. Elevated sympathetic tone has been shown to correlate with severe hypotension after SA. The aim of this prospective trial was to investigate skin conductance (SC), as a measure of sympathetic tone, to predict hypotension after SA. METHODS: After ethical approval and written informed consent, 30 patients undergoing SA were included. Baseline measurements of SC [number of fluctuations per second (reflecting the firing rate of skin sympathetic nerves), area under the curve (AUC) A and B (reflecting the magnitude of the sympathetic impulse)], blood pressure and heart rate were recorded. After administration of SA, all parameters were assessed every 2.5 min for a total of 15 min. Baseline readings of SC were compared with the lowest blood pressure within the study period. RESULTS: Data from 30 subjects [73 (8) years] were analysed. After SA, the mean arterial blood pressure declined an average of 21.3 (11.3) mmHg. A cut-off value of 0.35 microSs for baseline AUC B allowed prediction of more than mild hypotension (>15% from baseline) after SA with a sensitivity of 72.5% and a specificity of 77.5%. CONCLUSIONS: AUC B, as a parameter of SC, may predict severe arterial hypotension after SA in the elderly.

New parameters of skin conductance compared with Bispectral Index monitoring to assess emergence from total intravenous anaesthesia.

BACKGROUND: Arousal after total i.v. anaesthesia (TIVA) has been reported to be detectable by monitoring the number of fluctuations per second (NFSC), a parameter of skin conductance (SC). However, compared with monitoring of the bispectral index (BIS), the predictive probability of NFSC was significantly lower. The aim of this study was to determine the value of the two new, not yet published parameters of SC, area under the curve (AUC) methods A and B, for monitoring emergence from TIVA compared with monitoring of NFSC and BIS. METHODS: Twenty-five patients undergoing surgery were investigated. NFSC, AUC A, AUC B, BIS, and haemodynamic parameters (mean arterial pressure and heart rate) were recorded simultaneously. The performance of the monitoring devices in distinguishing between the clinical states 'steady-state anaesthesia', 'first clinical reaction', and 'extubation' were compared using the method of prediction probability (Pk) calculation. RESULTS: BIS showed the best performance in distinguishing between 'steady-state anaesthesia' vs 'first reaction' (Pk BIS 0.95; NFSC 0.73; AUC A 0.54; AUC B 0.62) and 'steady-state anaesthesia' vs 'extubation' (Pk BIS 0.99; NFSC 0.73; AUC A 0.71; AUC B 0.67). However, the time from first BIS>60/SC>0 to a first clinical reaction was significantly shorter for BIS (median BIS((R)) 180 s; NFSC 780 s; AUC A 750 s; AUC B 690 s; P < 0.001). CONCLUSIONS: AUC A and AUC B did not improve accuracy of SC monitoring in patients waking after TIVA.

Necrotizing fasciitis: a deadly infection.

Necrotizing fasciitis (NF) is a life-threatening condition, consisting of a soft-tissue infection with rapidly progressive, widespread fascial necrosis. NF may be caused by a wide variety of microbes. Indeed, NF may be an infection of one species of bacteria or may be polymicrobial. Prompt diagnosis and treatment are essential. Surgical debridement and antibiotic therapy are the primary treatment options.

Rocky Mountain spotted fever.

Rocky Mountain spotted fever (RMSF) is an unusual but important dermatological condition to identify without hesitation. The classic triad of headache, fever, and a rash that begins on the extremities and travels proximally to involve the trunk is found in a majority of patients. The cutaneous centripetal pattern is a result of cell to cell migration by the causative organism Rickettsia rickettsii. Such individuals should receive prompt antimicrobial therapy and supportive care to avoid serious and potentially fatal complications.

Pyoderma vegetans.

BACKGROUND: Pyoderma vegetans is a rare condition that is clinically characterized by large verrucous plaques with elevated borders and multiple pustules. The etiology of this disorder remains unknown. OBJECTIVES: We describe a 24-year-old woman with rapidly evolving pyoderma vegetans. Our patient had the unique additional findings of a highly elevated serum IgE level and a history of hidradenitis suppurativa. CONCLUSIONS: Pyoderma vegetans is diagnosed on clinical and histological criteria. Differentiation must be made from disorders such as pyoderma gangrenosum, Sweet's syndrome, and deep fungal infections. We illustrate a case of pyoderma vegetans and review the literature on this rare disorder. Clinical and histological criteria for diagnosis are presented, as well as differentiation from some mimicking disorders.

Tuberculous gumma (cutaneous metastatic tuberculous abscess) with underlying lymphoma.

Cutaneous tuberculosis is an infrequent first sign of disseminated tuberculosis. We describe a patient with 2 cutaneous ulcerations that grew Mycobacterium tuberculosis. Despite an initial response to antimycobacterial therapy, the fever relapsed. After several months, biopsy of a single cervical lymph node showed a T cell-rich B cell lymphoma. Our patient had metastatic tuberculous abscesses (tuberculous gummas), which are secondary to disseminated tuberculosis, and an underlying occult lymphoma, both believed to be sequentially presenting as a fever of unknown origin.

Bacillary angiomatosis in an HIV seronegative patient on systemic steroid therapy.

Bacillary angiomatosis is an unusual systemic vascular proliferation seen predominantly in patients with the acquired immunodeficiency syndrome. These vascular lesions are due to infection with a Bartonella species, most commonly B. henselae, but sometimes B. quintana. It is treatable and often curable, but without therapy may be life-threatening. Clinically, the disorder often resembles several different vascular disorders, particularly pyogenic granuloma and Kaposi's sarcoma. We now report a clinically typical patient with bacillary angiomatosis who was HIV seronegative, but who had idiopathic thrombocytopenic purpura, was status-post splenectomy and to whom long-term systemic prednisone had been administered.

Overwhelming postsplenectomy infection.

The incidence of overwhelming sepsis in asplenic patients is low. It carries, however, an unacceptably high morbidity rate and mortality rate. The risk of development is related to the age of the patient as well as the duration since and the reason for splenectomy. It may also occur in functionally hyposplenic states. The rapidly lethal clinical course demands intense efforts toward prevention by both physician and patient, rapid recognition within the initial medical setting, and aggressive intervention with broad-spectrum antibiotics and other supportive measures.