Withania somnifera or Ashwagandha is a medicinal herb of Ayurveda. Though the extract and purified molecules, withanolides, from this plant have been shown to have different pharmacological activities, their effect on bone formation has not been studied. Here, we show that one of the withanolide, withaferin A (WFA) acts as a proteasomal inhibitor (PI) and binds to specific catalytic β subunit of the 20S proteasome. It exerts positive effect on osteoblast by increasing osteoblast proliferation and differentiation. WFA increased expression of osteoblast-specific transcription factor and mineralizing genes, promoted osteoblast survival and suppressed inflammatory cytokines. In osteoclast, WFA treatment decreased osteoclast number directly by decreasing expression of tartarate-resistant acid phosphatase and receptor activator of nuclear factor kappa-B (RANK) and indirectly by decreasing osteoprotegrin/RANK ligand ratio. Our data show that in vitro treatment of WFA to calvarial osteoblast cells decreased expression of E3 ubiquitin ligase, Smad ubiquitin regulatory factor 2 (Smurf2), preventing degradation of Runt-related transcription factor 2 (RunX2) and relevant Smad proteins, which are phosphorylated by bone morphogenetic protein 2. Increased Smurf2 expression due to exogenous treatment of tumor necrosis factor α (TNFα) to primary osteoblast cells was decreased by WFA treatment. This was corroborated by using small interfering RNA against Smurf2. Further, WFA also blocked nuclear factor kappa-B (NF-kB) signaling as assessed by tumor necrosis factor stimulated nuclear translocation of p65-subunit of NF-kB. Overall data show that in vitro proteasome inhibition by WFA simultaneously promoted osteoblastogenesis by stabilizing RunX2 and suppressed osteoclast differentiation, by inhibiting osteoclastogenesis. Oral administration of WFA to osteopenic ovariectomized mice increased osteoprogenitor cells in the bone marrow and increased expression of osteogenic genes. WFA supplementation improved trabecular micro-architecture of the long bones, increased biomechanical strength parameters of the vertebra and femur, decreased bone turnover markers (osteocalcin and TNFα) and expression of skeletal osteoclastogenic genes. It also increased new bone formation and expression of osteogenic genes in the femur bone as compared with vehicle groups (Sham) and ovariectomy (OVx), Bortezomib (known PI), injectible parathyroid hormone and alendronate (FDA approved drugs). WFA promoted the process of cortical bone regeneration at drill-holes site in the femur mid-diaphysis region and cortical gap was bridged with woven bone within 11 days of both estrogen sufficient and deficient (ovariectomized, Ovx) mice. Together our data suggest that WFA stimulates bone formation by abrogating proteasomal machinery and provides knowledge base for its clinical evaluation as a bone anabolic agent.
osteoporosis; proteasome; bone formation; withaferin; drill hole; bone healing; micro-CT
Nonunion of intertrochanteric fractures is uncommon because there is excellent blood supply and good cancellous bone in the intertrochanteric region of the femur. A diagnosis of primary intertrochanteric nonunion is made when at least 15 weeks after the fracture there is radiological evidence of a fracture line, with either no callus (atrophic) or with callus that does not bridge the fracture site (hypertrophic). There is only one published series that exclusively describes seven primary nonunions of intertrochanteric fractures. The aim of the present study was to analyze the results of internal fixation, valgization with 135° dynamic hip screw (DHS), and bone grafting in patients with primary nonunion of intertrochanteric fractures.
Materials and Methods:
Eighteen patients with primary intertrochanteric nonunion were included in the study; 16 were male and 2 were female. The age range was 30–70 years (mean: 46.9 years). The mean duration since index injury was 8.5 months (range: 4–18 months). As per the AO classification, the fractures were 31A 1.1 (n=1), 1.2 (n=1), 2.2 (n=3), 2.3 (n=9), and 3.3 (n=4). Three patients had hypermobile nonunion and 15 had stiff nonunion. The surgical principle was excision of pseudarthrosis, if present (n=3); freshening of the bone ends; stable fixation with 135° DHS, with good proximal purchase; bone grafting; and valgization.
Union was achieved in all patients at an average of 5.62 months (range: 4–7 months). The Harris hip score improved from 38 points preoperatively to 86 postoperatively at healing. The average limb shortening improved by 2 cm (range: 1.5 cm–3 cm). There was no infection and pain at the hip at final follow-up in any of the cases. All patients were subjectively satisfied with the outcome. All were capable of full weight bearing on their affected limb.
Union in primary nonunion of intertrochanteric fractures in physiologically young patients with a well-preserved femoral head and good bone stock can be achieved with internal fixation, valgization, and grafting procedures.
Bone grafting; intertrochanteric fractures; valgization
Neurological syndromes are not an uncommon presentation with insulinomas. Recurrent hypoglycemia associated with it can present with a variety of neurological symptoms that may include disturbances of consciousness, seizures, stroke-like presentation, movement disorder, dementia and chronic neuropathy. The myriad of presentations, resemblance with other neurological conditions and episodic nature often lead to misdiagnosis and a delay in definitive treatment. Rare cases of insulinoma presenting as combination of abnormal movements have been described. We report a patient who presented with both hypoglycemia induced symptomatic seizures and paroxysmal non-kinesiogenic dystonic choreoathetosis. Insulinoma is a potentially treatable disorder and early definitive intervention can prevent long term neurological disability in patients.
Hypoglycemia; insulinoma; movement disorder; seizures
Three Vibrio species from the resident microflora of gastrointestinal tract of freshwater carps and prawns were isolated and confirmed biochemically as V. fluvialis from Cyprinus carpio/Labeo rohita; V. parahaemolyticus from Macrobrachium rosenbergii and V. harveyi from Macrobrachium malcomsoni. The genetic relationship among these Vibrio species was carried out by polymerase chain reaction (PCR) amplification of 16S rRNA gene followed by restriction digestion with Hae III, Bam HI and Pst I. Dendogram based on ribotyping showed the isolated Vibrios were differentiated into three clusters. V. harveyi was closely related to V. vulnificus (reference Microbial type Culture Collection (MTCC) strain) and distantly related to V. parahaemolyticus as well as V. fluvialis.
Freshwater Vibrios; 16S rRNA gene; Ribotyping
Life style related behavioural risk factors are mainly implicated for increased burden of cardio- vascular diseases. Research related to these risk behaviours especially among medical students is essential, considering their role as future physicians and role models in public health intervention programmes.
To evaluate the burden of cardiovascular risk behaviours among students of a medical college of Delhi, India.
Materials and Methods:
A cross sectional study was carried out among undergraduate medical students of a medical college in Delhi. Self administered questionnaire was used to collect information on identification data and risk behaviours in relation to cardiovascular diseases. Binary logistic regression analysis was done to calculate adjusted odds ratio to assess association between risk behaviours and covariates.
The minimum recommendation of taking at least five servings per day of fruits and vegetables was complied only by 12% of students. Consumption of carbonated soft drinks either once or more on daily basis was present in 23.7% students and 32.0% reported frequent consumption of fast foods in past week. Consumption of alcohol was present in 28.8% students but only small proportion of students (7%) was current tobacco users. Large proportions of students (42.6%) were either not carrying out or were involved in only occasional physical activity in past week.
Unhealthy behavioural practises are present and may progress as student advance through medical college. Developing strategies targeting at these risk behaviours and determining factors is necessary to promote healthy life style among medical students.
Cardio vascular; risk behaviors; medical college; students
A simple, accurate and economic spectrophotometric method for the determination of aripiprazole in tablet formulation is proposed. In the present method acidic solution of the aripiprazole formed colored ion-association complexes with bromocresol green, soluble in chloroform. Yellowish orange chromogen showed λmax at 414 nm and obeyed Beer's law in the concentration range of 10-60 μg/ml. Statistical analysis and recovery studies validated the method. The proposed method is rapid, precise and accurate and can be applied for the routine estimation of aripiprazole in the laboratory.
Aripiprazole; UV/Vis spectrophotometry
Apabahuka is a disease that usually affects the Amsa sandhi (shoulder joint).It is produced by the Vata dosha. Even though the term Apabahuka is not mentioned in the nanatmaja Vata vyadhi, Acharya Sushruta and others have considered Apabahuka as a Vataja vikara. Amsa shosha (wasting of the shoulder) can be considered as the preliminary stage of the disease, where loss or dryness of sleshaka kapha from amsa sandhi occurs. For the present study, Marsha nasya with Laghumasa Taila was administered to 15 patients for seven days, and the following results were obtained. After treatment, 53.33% relief was found on Bahupraspandita hara, 26.66% on Shoola, 30.00% on Stambha, 60.00% on Atopa, and 37.50% on wasting of muscles. On the overall effect of therapy alone, one (6.60%) patient got marked improvement, eight (53.33%) got moderate improvement, four (26.66%) were improved, and two (13.33%) patients remained unchanged.
Apabahuka; Marsha nasya; Laghumasa Taila; Vata vyadhi; Amsa shosha; Abhyanga; Swedana; Snehapana
Mycobacterium tuberculosis (MT) is a serious, but rare infectious complication after allogeneic bone marrow transplantation (BMT). We describe a case of fatal sepsis due to MT and Aspergillus flavus after allogeneic BMT for Aplastic Anemia. The diagnosis was made on bone marrow biopsy and asitic fluid culture. Broadspectrum antituberculous and Amphotericin B therapy was started immediately after diagnosis. The patient developed severe hypoxia and finally died of multi-organ failure. Rapid progression of mycobacterial infection as well as fungal infection should be considered in patients post BMT with unexplained fever, particularly in patients from endemic areas.
Allogenic BMT; Mycobacterium tuberculosis; Aspergillus flavus
A series of 1,2,4-dithiazole were synthesized from 1,2,4-thiadiazoles in the presence of CS2 and evaluated for their antimicrobial, anticonvulsant, analgesic and neurotoxicity potential. The compounds provided significant protection against maximal electroshock-induced seizures and seizures induced by 300 mg/kg of subcutaneous pentylenetetrazole administration. The designed compounds (3a-g) were screened in vitro for antibacterial activity against Staphylococcus aureus Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa and antifungal activity in fungal strains of Candida albicans and Aspergillus niger. Synthesized compounds exhibited moderate antibacterial and antifungal activity. N,N -Di-naphthalen-1-yl-N -(thioxo-5H -[1,2,4]dithiazol-3-yl)-guanidine and N,N -Bis-(4-fluoro-phenyl)-N -(5-thioxo-5H -[1,2,4]dithiazol-3-yl)-guanidine showed analgesic activity by tail flick method.
1,2,4-Thiadiazoles; 1,2,4-dithiazole; antimicrobial; anticonvulsant; analgesic; neurotoxicity
Certain 2-(1'-iminothioimido substituted)-1'-substituted phenybenzoic acids (P1-9) were synthesized by reaction of phthalic anhydride with benzotriazole, 2-mercapto benzothiazole and 2-p-amino phenyl benzimidazole, respectively (A1-3) followed by imine formation with Schiff bases of thiourea with salicylaldehyde, furfuraldehyde and 1-phenyl-3-methyl-5-pyrazolone. Antiulcer activity was evaluated using reduction in total acidity, free acidity and ulcer index as parameters. Compounds P3, P6, P7 and P9(100 mg/kg) showed significant (P< 0.001) antiulcer action compared to control and omeprazole (40 mg/kg).
Benzothiazole; benzotriazole; benzimidazole; total acidity; free acidity; ulcer index
New bidentate or tridentate Schiff bases and their
VO(II) and Co(II) complexes formed by the condensation of methyl
isobutyl ketone with nicotinamide (mna)/2-amino-4-chlorophenol
(map) and 2-hydroxy acetophenone with nicotinamide
(han)/isoniazide (hai). Physicochemical characterization has been
carried out to determine the structure of the complexes. The FAB
mass and thermal data show degradation pattern of the complexes.
XRD analysis reveals that all the studied complexes crystallize as
tetragonal crystal system. Some of the complexes have been
screened for their antimicrobial activity by the well diffusion
technique using DMSO as solvent on different species of pathogenic
bacteria/fungi, that is, E. coli, S. aureus, S. fecalis,
A. niger, T. polysporum, and their antimicrobial potency
have been discussed. It has been found that all the complexes are
antimicrobially active and show higher activity than the free
ligand. Metal chelation affects significantly the
antimicrobial/bioactive behavior of the organic ligands.
Antifungal activity of 2-hydroxy 4,4'6'trimethoxy chalcone individually was tested against spore germination of ten fungi of different genera. Efficacy of the chemical was also tested against conidial germination and other growth parameters of Erysiphe pisi on excised pea leaves. 2-Hydroxy 4,4'6'trimethoxy chalcone inhibited spore germination at all the concentrations. Maximum inhibition was observed at 2000 ppm where more than 78 per cent inhibition of spore germination was observed in Ustilago cynodontis, Alternaria brassicicola, A. solani and Aspergillus flavus. It also reduced conidial germination of E. pisi significantly, when applied as pre-inoculation treatment.
2-Hydroxy 4,4'6'Trimethoxy chalcone; Spore germination inhibition
Rhinosporidiosis is a chronic infestation by the fungus Rhinosporidium seeberi, which predominantly affects the mucus membranes of the nose and nasopharynx. We report a case of rhinosporidiosis with presentation as an oropharyngeal mass and a discusion about its endoscopic removal.
Endoscopic removal of rhinosporidiosis; nasal/oropharyngeal polypoidal mass; rhinosporidiosis
Cytokines play a key role in immune responses and inflammation. IL-1Ra is a naturally occurring structural variant of IL-1 that competitively inhibits receptor binding of IL-1. We have investigated the polymorphism in intron-2 of the interleukin-1 receptor antagonist gene in North Indian population. This genetic variation has been of great interest due to its possible association with a variety of human diseases primarily of epithelial and endothelial cell origin such as urolithiasis etc. Allele frequencies of the IL-1Ra polymorphism vary among different populations but there is no data till date reported from India. The present study was carried out to determine the IL-1Ra gene Polymorphism in 165 normal unrelated individuals from North India. We obtained an allelic frequency of 63.94, 30.61, 4.55, 0.90 for A, B, C and D allele and percentage of genotypes AA, BB, CC, DD, A/B, A/C, A/D and B/C were 49.7, 18.2, 2.42, 0.60, 24.2, 3.63, 0.60, 0.60 respectively. Our results suggested that the frequency and distribution of this polymorphism in India is substantially different from other populations and ethnic groups.
IL-1 Ra Polymorphism; VNTR; Single nucleotide polymorphisms (SNPs)
Bullet; parapharyngeal space
The purpose of this study was to prepare conventional and sterically stabilized liposomes containing leuprolide acetate in an attempt to prolong the biological half life of the drug, to reduce the uptake by reticuloendothelial system (RES), and to reduce the injection frequency of intravenously administered peptide drugs. The conventional and sterically stabilized liposomes containing leuprolide acetate were prepared by reverse phase evaporation method and characterized for entrapment efficiency and particle size. Radiolabeling of leuprolide acetate and its liposomes was performed by direct labeling with reduced technetium-99m. Its biodistribution and imaging characteristics were studied in ehrlich ascites tumor (EAT)-bearing mice after labeling with technetium-99m. The systemic pharmacokinetic studies were performed in rabbits. A high uptake by tumor was observed by sterically stabilized liposome containing leuprolide acetate compared with free drug and conventional liposomes. The liver/tumor uptake ratio of free drug, conventional (LL), and sterically stabilized liposomes (SLL5000 and SLL2000) was found to be 20, 7.99, 1.63, and 1.23, respectively, which showed the increased accumulation of sterically stabilized liposomes in tumor compared with the free drug and conventional liposomes at 24 hours postinjection. Liver uptake of sterically stabilized liposomes was still 7-fold less than the conventional liposomes. The marked accumulation of liposomes in the tumor-bearing mice was also documented by gamma scintigraphic studies. The findings demonstrate the distribution of these liposomes within solid tumor and prove that the sterically stabilized liposomes experience increased tumor uptake and prolonged circulation half life. Hence these findings will be relevant for the optimal design of long circulating liposomes for the peptide drugs and for targeting of liposomes toward tumor.
sterically stabilized liposomes; leuprolide acetate; technetium-99m; biodistribution; gamma imaging
Bullet; Parapharyngeal space
A 12-year-old girl presented with complaint of passing very ‘scanty’ amount of urine, approximately 50 ml every alternate day for the last five years. She had been admitted for investigations several times in different hospitals in the past. On evaluation, she was found to have no abnormality related to the urinary tract. Detailed psychiatric assessment revealed the factitious nature of her symptoms, suggesting the diagnosis of factitious disorder with physical symptoms.
Factitious disorder; Munchausen's syndrome; oliguria; oligoanuria