Molecular Characterization and Analysis of Risk Factors Associated with Brucellosis in Central Indian and Meghalaya Population.

Objective: In this study we evaluated the utility of Abortus Melitensis Ovis Suis Brucella PCR (AMOS PCR) for the molecular characterization of Brucella species and analyzed the associated risk factors for brucellosis in Central Indian and Meghalayan population. Methods: AMOS PCR was carried out in a total of 160 BSCP-31 PCR-positive DNA samples isolated previously from the blood of Central Indian (n = 90) and Meghalayan cohorts (n = 70). Clinical and associated risk factors recorded earlier were used to establish strain-specific disease outcomes in study cohorts. Results: Brucella melitensis was found to be the dominant strain in both Central Indian and Meghalayan cohorts (57.7% and 54.28%, respectively) followed by Brucella abortus (42.22% and 38.57%). Although rare, brucellosis cases in the Meghalayan population also showed the presence of Brucella suis (7.14%) and Brucella ovis (2.85%). Febrile illness was a major clinical risk factor in both study cohorts, while occupational risk factors like exposure to animals and raw milk consumption were major mediating factors for brucellosis in Central Indian cohorts. On the contrary, meat consumption was found to be significant predisposing factor for brucellosis in Meghalaya. Conclusion: Molecular characterization of Brucella species provides important public health data for mitigation, advocacy, and antimicrobial stewardship.

Diagnostic utility of LAMP PCR targeting bcsp-31 gene for human brucellosis infection.

PURPOSE: Human brucellosis is a neglected zoonotic disease of significant public health concern. Molecular diagnosis of brucella remains challenging in low resource settings, due to the high infrastructure and cost involved. Loop-mediated isothermal amplification (LAMP) is a rapid point of care polymerase chain reaction (PCR) with the utility of on-field molecular diagnosis and offers a convenient alternative to conventional PCR. In the present study, we developed and evaluated the diagnostic utility of in house LAMP PCR targeting the Brucella genus-specific bcsp-31 gene in patients having febrile illness. MATERIALS AND METHODS: The analytical sensitivity and specificity of bcsp-31 LAMP PCR was first evaluated using brucella (n â€‹= â€‹8) and non-brucella cultures (n â€‹= â€‹5), along with spiked clinical samples. The overall diagnostic utility of developed LAMP PCR was then further evaluated in 393 human samples suspected of brucellosis. RESULTS: The developed LAMP PCR could detect as low as 8 â€‹fg of DNA by visual detection within 35min. We report sensitivity and specificity of the developed LAMP PCR as 90.91% and 99.37%.The accuracy of the developed test assay was found to be 98.60%. In clinical samples, LAMP gave positivity of 20% with the concordance of 89% with conventional PCR. CONCLUSION: To conclude, a rapid, efficacious, sensitive LAMP PCR targeting the bcsp 31 gene was developed. The existing LAMP PCR can be used as a point of care screening test in various low resource endemic setting in lieu of conventional PCR for estimation of prevalence data, diagnosis and treatment of brucellosis.

Therapeutic Potential of Dillenia indica L. in Attenuating Hyperglycemia-Induced Oxidative Stress and Apoptosis in Alloxan-Administered Diabetic Mice.

BACKGROUND: Hyperglycemia-induced oxidative stress accelerates the process of apoptosis in tissues. Dilleniaindica (DI) is a medicinal plant, and its fruit contains many therapeutic properties. The therapeutic activity of the Methanolic Fruit Extract (MFE) of DI in attenuating oxidative stress and apoptosis in the liver and kidney tissues of alloxan-induced diabetic mice was analyzed in the present study. METHODS: High-Performance Thin Layer Chromatography (HPTLC) profiling of MFE was conducted. GLUT4 protein expression analysis and lipid peroxidation assays were conducted to check for MFE effect by administering in diabetic mice. An ultrastructural study was conducted for both the tissues. In apoptotic studies, the TUNEL assay and apoptotic protein expression analysis was conducted. RESULTS: High-Performance Thin Layer Chromatography (HPTLC) profiling of MFE showed the presence of two crucial antioxidants, ascorbic acid, and naringenin. In GLUT-4 protein expression analysis, MFE suppresses hyperglycemia by upregulating GLUT4 protein expression. Lipid peroxidation assay showed a decrease in malondialdehyde (MDA) upon MFE administration in diabetic mice. An ultrastructural study was conducted, and MFE was found to restore cellular alterations in diabetic tissues. In apoptotic studies, the TUNEL assay shows that MFE treatment showed fewer apoptotic cells than the diabetic group. The study also observed decreased caspase 3 protein expression and increased Bcl-2 protein expression. CONCLUSIONS: Therefore, it is inferred from the study that MFE can exert a protective effect by suppressing hyperglycemia and modulating oxidative stress and apoptosis in alloxan-administered diabetic mice.

Seroepidemiological study of human brucellosis in the Northeast region of Meghalaya, India.

Introduction: Brucellosis is a neglected zoonotic disease of major public health concern. In India, the incidence of brucellosis remains vastly underreported due to its non-specific clinical presentation and sub-optimal sensitivity of existing gold standard tests. Studies in Northeast India have shown high incidences of brucellosis in livestock, but the region lacks data on human brucellosis despite its high associated risk. In the present study, we report the seroprevalence of human brucellosis and its associated risk factors in Meghalaya, Northeast India. Materials and Methods: A prospective observational study was conducted in East Khasi Hills and Ri.Bhoi districts of Meghalaya, from July 2018 to July 2020. A total of 1046 suspected patients with febrile illness along with associated risk factors were recruited through camps and various diagnostic laboratories in the defined region as per the pre.specified inclusion and exclusion criteria. Baseline, demographics, and clinical characteristics were recorded of all the consenting participants. Blood samples were analyzed for brucellosis-specific IgM antibodies through enzyme-linked immunosorbent assay (ELISA). Results and discussion: The overall seroprevalence of brucellosis was found to be 11.37% in Meghalaya. Among recruited participants, females were found to be more susceptible than males. Risk factors such as consumption of meat were found to be more significantly associated with brucellosis disease in the study region. Among the clinical presentations, pyrexia of unknown origin, myalgia, and chronic fatigue syndrome were found to be significantly associated with brucellosis disease in IgM.positive cases. Conclusion: Our result suggests further epidemiological investigations for human brucellosis in Northeast India toward improved advocacy for accurate diagnosis, and development of proper response mechanism in areas of high endemicity.

Bioactive Khadi Cotton Fabric by Functional Designing and Immobilization of Nanosilver Nanogels.

The antimicrobial finishing is the most suitable alternative for designing medical textiles for biomedical applications. The present investigation aims at the preparation of skin-contacting khadi cotton fabric that would prevent microbial infection and offer excellent skin compatibility. A simple approach has been followed for the preparation of bioactive nanogels for antimicrobial finishing of the khadi cotton fabric. Bioactive nanogels were synthesized by using aloe vera (AV) as a reducing agent for silver ions in the presence of polyvinyl alcohol (PVA). PVA stabilizes the growth of silver nanoparticles, which is influenced by the variation in the reaction time and the temperature. Nanogels were characterized by transmission electron microscopy and scanning electron microscopy analyses. The nanogels exhibited strong antimicrobial behavior against both Staphylococcus aureus and Escherichia coli, as confirmed by the colony count method. Almost 100% antibacterial behavior was observed for the nanosilver content of 10 mM. The nanogel-finished khadi fabric showed bactericidal properties against both S. aureus and E. coli. The nanogel-finished fabric exhibited high hydrophilicity allowing complete water droplet penetration within 10 s as compared to 136 s in virgin fabric. Moreover, the skin irritation study of the fabric on male Swiss albino mice did not show any appearance of dermal toxicity. These results demonstrated that the bioactive finished khadi fabric is appropriate as skin contacting material in human health care.

Chlorophyllin supplementation modulates hyperglycemia-induced oxidative stress and apoptosis in liver of streptozotocin-administered mice.

Chlorophyllin is a water-soluble mixture of sodium-copper salts of chlorophyll with antioxidant and antimutagen properties. In this study, an attempt has been made to evaluate the effect of chlorophyllin on hyperglycemia-induced oxidative stress and apoptosis in liver of streptozotocin (STZ)-administered mice. In STZ-induced diabetes, two causative factors for pancreatic ß-cell deaths are DNA alkylation and profound reactive oxygen species (ROS) generation. In this study, chlorophyllin treatment was found to be able to modulate oxidative stress and apoptosis in liver of diabetic mice. Diabetic mice exhibited a significant reduction of ROS, malondialdehyde (MDA), and protein carbonyl levels upon treatment with the chlorophyllin. However, antioxidant enzymes, such as copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), and catalase (CAT) showed enhanced activity as well as expression in chlorophyllin-administered diabetic mice. The hepatoprotective effect of chlorophyllin was confirmed from the decreased activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The histological and ultrastructural studies revealed the ability of chlorophyllin to restore morphological and cellular alterations as observed in STZ-induced diabetic mice. The effect of chlorophyllin on apoptosis showed the downregulation of cysteine-dependent aspartate-specific protease (caspase) 3 and caspase 9, whereas upregulation of B-cell lymphoma-2 (Bcl-2) protein, and the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) assay demonstrated a few apoptotic cells. In conclusion, it can be stated that chlorophyllin treatment can exert hepatoprotective effect via modulating hyperglycemia-induced oxidative stress and apoptosis in STZ-administered diabetic mice. © 2018 BioFactors, 44(5):418-430, 2018.

Development of novel wound care systems based on nanosilver nanohydrogels of polymethacrylic acid with Aloe vera and curcumin.

This study is aimed at the development of a composite material for wound dressing containing nanosilver nanohydrogels (nSnH) along with Aloe vera and curcumin that promote antimicrobial nature, wound healing and infection control. Nanosliver nanohydrogels were synthesized by nanoemulsion polymerization of methacrylic acid (MAA) followed by subsequent crosslinking and silver reduction under irradiation. Both the polymerization and irradiation time had significant influence on the nanoparticle shape, size and its formation. Polyvinyl alcohol/polyethylene oxide/carboxymethyl cellulose matrix was used as gel system to blend with nSnH, A. vera, curcumin and coat it on the hydrolysed PET fabric to develop antimicrobial dressings. The cumulative release of silver from the dressing was found to be ~42% of the total loading after 48h. The antimicrobial activity of the dressings was studied against both Staphylococcus aureus and Escherichia coli. In vivo wound healing studies were carried out over a period of 16d on full-thickness skin wounds created on Swiss albino mice. Fast healing was observed in Gel/nSnH/Aloe treated wounds with minimum scarring, as compared to other groups. The histological studies showed A. vera based dressings to be the most optimum one. These results suggest that nSnH along with A. vera based dressing material could be promising candidates for wound dressings.

A study on evaluation of apoptosis and expression of bcl-2-related marker in wound healing of streptozotocin-induced diabetic rats.

Uncontrolled blood sugar is a major cause of vascular complications and delayed wound healing in diabetes mellitus. During wound healing process, normally, apoptosis is responsible for events such as removal of inflammatory cells and evolution of granulation tissue into scar which occur during the late phase of wound healing. Early apoptosis can lead to abnormal wound healing by removing granulation tissue including fibroblast, endothelial cell, and small vessels. To determine the role of apoptosis in association with hyperglycemia in diabetic wound healing, apoptosis-related intracellular marker such as expression of Bcl-2 protein by immunohistochemistry and normal histology has been studied. Histological findings show higher level of apoptosis and diminished granulation tissue formation in diabetic rats wounds along with minimal expression of Bcl-2 in diabetic rats wounds when compared with nondiabetic rats wounds. It can be concluded from this study that elevated blood sugar level may be associated with increased apoptosis and the least expression of Bcl-2 protein which might cause deregulation of the wound healing processes in streptozotocin-induced diabetic rats.

Development of a novel rapid immunodiagnostic kit based on flagellar 40 kDa antigen epitope for the detection of typhoid fever in Indian patients.

To aid the clinical diagnosis of typhoid fever in India, where most hospitals and primary health centres have no facilities for culture, we report on the development of a novel and rapid immunodiagnostic kit for the direct detection of Salmonella Typhi--specific IgG antibodies against S. Typhi flagellar H antigen. The disease often does not show a specific clinical picture, and can be confused with other febrile illness such as malaria, dengue fever and Staphylococcus aureus. To overcome the problem of cross reactivity specific epitope of the flagellar H antigen was immobilised on the testing kit strip eliminating chances of cross reactivity and false positive results thereby increasing the specificity of the test. Since the immunodiagnostic kit, uses the flagellar H antigen from bacteria present in our country, the antibodies present in the serum of patients of our country will have maximum binding affinity, enhancing the sensitivity of our test kit. The immunodiagnostic kit on analysis gave a positive result with clinically diagnosed typhoid positive patient serum and negative results were obtained with the sera of clinically diagnosed malaria, abscess of Staphylococcus aureus and Visceral leishmaniasis (Kala-azar) patients.

Correlation between IL-7 and MCP-1 in diabetic chronic non healing ulcer patients at higher risk of coronary artery disease.

Cytokines play an extremely important role in the pathogenesis of coronary artery disease (CAD) in which interleukin (IL)-7 is a major regulator of T-cell homeostasis which is conced in the stimulation of leukocyte-endothelial cell adhesion during inflammatory events. Circulating IL-7 is associated with activation of monocyte and natural killer cells, leading to enhanced production of inflammatory cytokines and chemokines observed in atherosclerosis and acute coronary syndromes. Plasma levels of IL-7, hs-CRP and monocyte chemoattractant protein (MCP)-1 were measured by an immunoenzymatic ELISA technique. Ninety neuropathic diabetic foot patients were divided into two groups: group B [those without CAD (n=45)] and group C [those with higher risk of CAD (n=45)]. Thirty-five healthy subjects were included as control (group H). Plasma concentration of IL-7, MCP-1 and hs-CRP were significantly higher in group C as compare with group H and B. Plasma IL-7 levels also showed significant positive correlations with plasma levels of hs-CRP and MCP-1. Abnormalities in lipid profile were also observed. In conclusion the positive correlation between plasma concentration of IL-7, MCP-1 and hs-CRP in diabetic foot patients observed herein, suggests a plausible role for IL-7 in the promotion of clinical instability in coronary artery disease.

Acetabular reconstruction using fresh frozen bone allograft.

The success of total hip arthroplasty has led to a move toward operating on a wider and younger population. All implants have a finite useful life, and bone loss is associated with all major causes of failure. The use bone allograft, either morselized or structural has the aim of reconstituting the bone stock. We retrospectively reviewed the clinical and radiological results of 72 acetabular reconstructions both primary and revision, done at our institute between May 1999 and October 2004 with a minimum follow-up of one year. The Harris Hip Score (HHS) was calculated and follow-up radiographs were evaluated for graft incorporation; evidence of loosening and migration. Preoperatively 28 acetabular defects were type III (AAOS), 18 were type II, 16 were type I and 10 were type IV. 60 reconstructions were done using cementless acetabular prostheses, and the remaining twelve had cemented implants. Patients were followed up for an average of 30.69 months (range 12 - 64 months). The mean preoperative HHS was 36.13 points which improved to a mean of 81.6 points (range 61 to 91) at final follow-up. None of the patients required reoperation. Dislocation was the commonest complication (8 hips). Short and medium term results were encouraging with the use of this technique.

Comparison of total knee arthroplasty in stiff knees and knees with good preoperative range of motion.

This study compared the results of total knee arthroplasty (TKA) in patients with stiff knees and patients with good preoperative range of motion. Results following TKA were compared in 30 patients (52 knees) with severely degenerated knees with preoperative range of motion < 50 degrees (group 1) and 30 patients (50 knees) who were matched for age, gender, and diagnosis with preoperative range of motion > 90 degrees (group 2). Results were evaluated for range of motion and Knee Society score, as well as the need for extensile approaches, soft-tissue releases, additional bone cuts, and constrained prostheses. Radiographic results were evaluated for implant positioning, patellar height, and alteration in joint line. Findings indicated that although TKA in stiff knees can be successful, the results are inferior to those obtained in standard TKA. Careful preoperative planning and meticulous surgical technique are essential for good results.

Primary cementless total hip arthroplasty for bony ankylosis in patients with ankylosing spondylitis.

We retrospectively reviewed 54 patients (92 hips) who underwent cementless total hip arthroplasty for bony ankylosis in ankylosing spondylitis between September 1988 and 2002. The mean age of the patients was 25.5 years. The mean duration of follow-up was 8.5 years. The average preoperative Harris Hip Score of 49.5 improved to 82.6 postoperatively. Postoperatively, 10 hips had mild to moderate pain. Anterior dislocation occurred in 4 (4.3%) hips, and sciatic nerve palsy in 1 (1.1%) hip. Heterotopic ossification was seen in 12 patients; reankylosis rate was 0%. Thirteen (14%) arthroplasties were revised because of aseptic loosening. Kaplan-Meier survivorship analysis with revision as end point revealed 98.8% survival at 5 years and 85.8% survival at 8.5 years follow-up.

Management of extensor mechanism deficit as a consequence of patellar tendon loss in total knee arthroplasty: a new surgical technique.

Extensor mechanism disruption is an uncommon but devastating complication of total knee arthroplasty. A new technique of extensor mechanism reconstruction for patellar tendon loss, after total knee arthroplasty, with the help of extensor mechanism composite allograft is described. Four patients with chronic extensor mechanism-deficient total knee arthroplasty were undertaken for revision surgery along with reconstruction of extensor mechanism with an innovative technique using an extensor mechanism composite allograft consisting of a patella-patellar tendon-tibial tubercle. On final follow-up, none of the patients had extensor lag but for 10 degrees of extensor lag in 1 patient only. Providing an environment for bone-to-bone healing both proximally as well as distally and supervised postoperative rehabilitation led to encouraging results in the management of a failed extensor mechanism after total knee arthroplasty.

Fracture dislocation of the spine after total hip arthroplasty in a patient with ankylosing spondylitis with early pseudoarthrosis.

BACKGROUND CONTEXT: There are no studies suggesting the management of asymptomatic, early pseudoarthrosis with advanced hip and spine deformities in patients with ankylosing spondylitis. Literature advocates the correction of the hip deformity first and that of spine later. PURPOSE: To highlight the importance of asymptomatic, early pseudoarthrosis lesion in a patient of ankylosing spondylitis with advanced hip and spine deformities. STUDY SETTING: A 25-year-old female patient suffering from ankylosing spondylitis with 70 degrees of flexion deformity at right hip and rounded kyphosis of thoracolumbar spine was admitted for right total hip arthroplasty. METHODS: Cementless total hip arthroplasty and flexor tendon release were performed on right hip under general anesthesia and patient was nursed in supine position postoperatively. RESULTS: The patient developed cauda equina syndrome and loss of sensation below twelfth thoracic segment on the second postoperative day. Radiological evaluation showed a fracture dislocation of the second and third lumbar vertebrae. Retrospective re-examination of preoperative radiographs showed small erosion of the anteroinferior margin of the second lumbar vertebra. Patient's neurological status did not improve after decompression and posterior stabilization of fractured spine. CONCLUSION: Careful evaluation of the spine in patients with spondyloarthropathy with advanced hip and spine deformities should be done. Any lesion that can potentially destabilize the spine should be treated first before correcting hip deformity.

Conversion of failed hemiarthroplasty to total hip arthroplasty: a short to mid-term follow-up study.

BACKGROUND: The conversion of hemiarthroplasty (unipolar or bipolar) of the hip to total hip replacement has been reported to be associated with very high rates of intra- and postoperative complications. We present a prospective analysis of the outcome of conversion surgery in patients with failed hemiarthroplasty. MATERIALS AND METHODS: Forty-four cases, 30 women and 14 men, average age 62 years (range 42-75 years) of failed hemiarthroplasty were converted to total hip replacement between January 1998 and December 2004. Groin pain was the main presenting complaint in the majority of the patients (24 out of 44). Six patients had infection and were operated with staged procedure. All acetabular and the majority (86.5%) of femoral components used in our series were uncemented. RESULTS: After an average follow-up of 6.4 years (range, two to nine years) Harris hip scores improved from 38 (range 15-62) preoperatively to 86 (range 38 to 100) and 22 (50%) patients were community ambulators without support while 17 (38%) needed minimal support of cane. Fifteen out of 18 (83%) patients who had isolated groin pain preoperatively experienced no pain postoperatively while three patients (17%) reported only partial improvement. Intraoperative and postoperative complications included iatrogenic fracture of the femur in two, femoral perforation in two, partial trochanteric avulsion in two, fracture of the acetabular floor in three hips, and postoperative dislocation in one. None of these complications resulted in a poor long-term outcome. The rate of loosening in our series was 2.3% (one out of 44) after a mean follow-up of 6.4 years with a mean survival of 97.4% at 72 months. CONCLUSION: Conversion of symptomatic hemiarthroplasty to total hip arthroplasty is a safe option that gives good functional results, with marginally higher rates of intra-operative complications. The patients should be warned of the possibility of incomplete relief of groin pain postoperatively.

Bilateral femoral neck stress fractures following total knee arthroplasty: a case report and review of literature.

Stress fracture of the femoral neck following total knee arthroplasty (TKA) is an uncommon complication with only 20 cases having been described in English literature so far. Stress fractures of femoral neck occurring simultaneously on both sides following a bilateral total knee replacement is an exceptional occurrence, which to the best of our knowledge, has not been described previously. We describe a patient suffering from osteoarthritis of both knees, who was treated with bilateral total knee arthroplasty simultaneously. He developed spontaneous fractures of femoral neck on both sides 9 months following the total knee replacement in the absence of any known risk factors for stress fractures. He was treated with bipolar hemi replacement arthroplasty for displaced fracture on one side and closed reduction/internal fixation on the undisplaced side. A high index of suspicion is needed to diagnose this unusual complication of total knee arthroplasty before the fracture displaces necessitating a prosthetic replacement.

Surgical management of intercondylar fractures of the humerus using triceps reflecting anconeus pedicle (TRAP) approach.

BACKGROUND: Operative fixation of intra-articular fractures of the distal humerus requires adequate exposure. The transolecranon approach is a commonly used approach. The olecranon osteotomy has potential complications related to prominence/migration of hardware, displacement/nonunion of osteotomy and triceps weakness. Triceps-reflecting anconeus pedicle (TRAP) approach avoids the olecranon osteotomy without compromising the operative exposure. We present outcome of fixation of displaced intra-articular distal humeral fractures with the use of TRAP approach. MATERIALS AND METHODS: We reviewed the functional and radiological results of 40 consecutive patients with intercondylar fractures of the humerus treated by internal fixation through TRAP approach. There were 28 males and 12 females and the average age was 32 ± 4.5 years. The right elbow was involved in 27 patients and the left elbow in 13 patients. The mechanism of injury was a fall in 20 patients, a motor-vehicle accident in 16 patients and direct trauma in four patients. RESULTS: At a minimum follow-up of 12 months (average 18 ± 4 months) 35 (87.5%) patients had good triceps strength. The average range of motion was 118.4 ± 7 degrees (range 80°-130°). The average time to union was 3.2 ± 1.6 months (range two to six months). No patient had triceps rupture, implant failure, neurovascular deficit or nonunion. Two patients needed removal of the implant because of subcutaneous prominence. CONCLUSIONS: The TRAP approach provides good visualization for fixation of intercondylar fractures of the humerus, without any noticeable untoward effect on triceps strength and postoperative rehabilitation; and one can avoid iatrogenic fracture of the olecranon and its associated complications.