Effect of temperature and chain length of added primary amines on the sphere to rod transition of aqueous ionic micelles

The importance of micelles as templates for nanomaterials is growing day by day. This resulted in an increasing interest for micelles in different sizes and shapes. Addition of n-amines to micellar solutions was found to bring change in the shape of the micelles from sphere to rod in aqueous ionic micellar solutions. The change in shape is qualitatively obtained from sudden change in the slope of pH versus amine concentration plots because the degree or protonation of n-alkylamines depends on the shape of micelles. In the present investigation, pH is measured at different temperatures to elucidate the influence of addition of n-amines on sphere-to-rod transition in aqueous micellar solutions. The surfactants employed in the present investigation are cetyltrimethyl ammonium bromide (CTAB), cetylpyridinium chloride (CPC), and sodium dodecyl sulphate (SDS).As the amine concentration is increased, the pH increases linearly at certain amine concentration and the slope of the resulting straight line changes on further addition of amine. It is noticed that increasing temperature requires more amine for structural transition of aqueous ionic micelles. It is also observed that the effectiveness of added amines leading to shape transition from sphere to rod is in the order of C8NH2>C7NH2> C6NH2.

Method Development And Validation Of Selective And Highly Sensitive Method For Determination Of Apixaban In Human Plasma Using Liquid Chromatography-Tandem Mass Spectrometry

Objective: The present research work aims to develop and validate a selective and highly sensitive method for the determination of apixaban in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods: 200 µl of sodium heparin plasma samples were acidified and clean-up was performed by using solid-phase extraction (SPE). Apixaban 13C D3 was used as an internal standard (deuterated) to lower the relative matrix effects and a single step SPE was employed for sample clean up. 10 µl of SPE eluent was loaded onto Hypersil Beta Basic C18, 100×4.6 mm, 5 µ column for highly selective chromatographic separation using an isocratic mobile phase. 2 mmol ammonium acetate in water and acetonitrile were delivered by using a quaternary low-pressure gradient pump without premixing at a minimum flow rate of 0.50 ml/min. Results: LC-MS/MS method was successfully developed and validated to demonstrate the lowest detection limit of 0.05 ng/ml and a linear dynamic range from 1-250 ng/ml with r2>0.99. Method development and validation results proved that the method is selective and highly sensitive for the determination of apixaban in human plasma using LC-MS/MS. Conclusion: Current method can be applied for both therapeutic drug monitoring (TDM) and pharmacokinetic (PK) study analysis.

Study of Ocular Manifestations among Beta Thalassemia Major Patients

Person with thalassemia major may have severe symptoms and may need regular blood transfusions. Iron overload is due to repeated blood transfusions and enhanced iron absorption by gastrointestinal tract and also creates negative impact on the organs function. We aimed to study the ocular changes in beta thalassemia major at a tertiary care hospital. Methods: In this prospective observational study, 234 patients diagnosed with beta thalassemia major, receiving multiple blood transfusions as a part of treatment were included. Patient brief history was taken, along with family history. Complete eye examination done. Ocular examination done by measuring visual acuity, refractive error assessment with autorefractometer, slit lamp examination, fundoscopy, perimetry, tonometry, color vision testing and tear break up time (TBUT) test. Results: A total of 234 beta thalassemia major patients of both sexes were evaluated, among them 132 (56.4%) were males and 102 (43.5%)were females. The mean age of thalassemic study population was 25.6±6.3. Most commonly observed were pinguecula (44%), visual field defects (40.5%), vascular tortuosity (39.3%), dry eye (33.3%), Refractive error (20.5%), Anterior segment involvement (18.8%), Cataract (13.2%), color vision defect (5.1%), normalization of optical vessels (4%). Conclusion: Regular Opthalmological examination helps to detect early changes due to disease and chelating agents. Issue of iron overload among thalassemic patients can be reduced by decreasing the need or the frequency of blood transfusions.

Obstacles Facing by Patients to Undergo Cataract Surgeries - A Hospital Based Study in and Around Ananthapuramu.

Background: Cataract estimated to be the cause of blindness among 17.6 million people worldwide. A few studies have done in India about obstacles or barriers of patients to undergo cataract surgeries, so the present study has undertaken to know the reasons for not accepting cataract surgical services. Methods: Patients in the age group of above 45 years, vision <20/60 and presenting with features of cataract were included. BCVA (Best corrected Visual acuity) was measured using Snellen chart and Cataract was diagnosed using torch light and Direct Ophthalmoscopy. Patients knowledge about cataract, factors obstructing patients to undergo cataract surgeries, regarding service delivery, cost and affordability were assessed. Results: Out of 120 patients, 46 (38.3%) were aware that they have cataract. Among various obstacles, about 60% were able to manage daily work and worry about the cost of surgery. Obstacles including Busy with work, cans see clearly with other eye were seen in above 50% of patients. Out of 120 patients, 83 (69.1%) did not undergo cataract surgery because of insufficient family income, whereas 54 (45%) persons does not have accompany to attend for cataract surgery.Conclusion: Need of eye health campaigns, providing services like, free of cost, transport facilities and place of treatment, follow up services. Many Government Organizations and Hospitals are conducting free camps for cataract surgeries, other than these health education is necessary by explaining about the beneficiaries of cataract surgery and advise people to avail health benefits or free camps to undergo cataract surgery.

Novel mutations of the arylsulphatase B (ARSB) gene in Indian patients with mucopolysaccharidosis type VI.

Background & objectives: Mucopolysaccharidosis type VI (MPS VI) is a rare, autosomal recessive lysosomal storage disorder caused by deficient enzymatic activity of N-acetyl galactosamine-4-sulphatase resulting from mutations in the arylsulphatase B (ARSB) gene. The ARSB gene is located on chromosome 5q11-q13 and is composed of eight exons. More than hundred ARSB mutations have been reported so far, but the mutation spectrum of MPS VI in India is still unknown. Hence, the aim of the present study was to identify the mutational spectrum in patients with MPS VI in India and to study the genotype-phenotype association and functional outcomes of these mutations. Methods: Molecular characterization of the ARSB gene by Sanger sequencing was done for 15 patients (aged 15 months to 11 yr) who were enzymatically confirmed to have MPS VI. Age of onset, clinical progression and enzyme activity levels in each patient were studied to look for genotype-phenotype association. Haplotype analysis performed for unrelated patients with the recurring mutation W450C, was suggestive of a founder effect. Sequence and structural analyses of the ARSB protein using standard software were carried out to determine the impact of detected mutations on the function of the ARSB protein. Results: A total of 12 mutations were identified, of which nine were novel mutations namely, p.D53N, p.L98R, p.Y103SfsX9, p.W353X, p.H393R, p.F166fsX18, p.I220fsX5, p.W450L, and p.W450C, and three were known mutations (p.D54N, p.A237D and p.S320R). The nine novel sequence variants were confirmed not to be polymorphic variants by performing sequencing in 50 unaffected individuals from the same ethnic population. Interpretation & conclusions: Nine novel mutations were identified in MPS VI cases from India in the present study. The study also provides some insights into the genotype-phenotype association in MPS VI.

Synthesis, characterization and use of Poly (N-isopropylacrylamide-co-N-vinylcaprolactam) crosslinked thermoresponsive microspheres for control release of Ciproflaxin hydrochloride drug.

Poly (N-isopropylacrylamide-co-N-vinyl caprolactam) copolymer was synthesized as an interesting thermoresponsive material possessing a phase transition temperature around 320C in phosphate buffer, pH 7.4 (PB). Thermoresponsive Poly(N-isopropylacrylamide-co-Nvinylcaprolactam) designated as PNIPA-NVC microspheres crosslinked with N,N’–methylene bisacrylamide (NNMBA) have been prepared by dispersion polymerization using varying amounts of NIPA, NVC and NNMBA., ciproflaxin hydrochloride (CFH) an anti- bacterial drug, was loaded into the microspheres during in situ polymerization and in vitro release of CFH has been studied. The microspheres were characterized by Fourier Transform Infrared Spectroscopy (FTIR) Differential Scanning Calorimetry (DSC), X-Ray Diffractometry (X- RD) and Scanning Electron Microscopy (SEM). The in-vitro release of CFH drug from the microspheres was studied in pH 7.4 medium, at the temperatures 250C & 370C. The microspheres consisting of NIPA and NVC provide thermo responsive nature to the microspheres. The system developed in this study showed a thermoresponsive for the controlled release of CFH. FTIR spectroscopy explained the formation of copolymer. The DSC and XRD techniques indicated the uniform distribution of drug in the microspheres. SEM studies indicated surface morphology of the microspheres. Prolonged and controlled release of CFH was achieved in a controlled manner upto 12 h.

Malignant peripheral nerve sheath tumor with divergent differentiation.

A malignant peripheral nerve sheath tumor (MPNST) is an uncommon spindle cell sarcoma accounting for approximately 5% of all soft tissue sarcomas. A 55-year-old female with a right suprarenal tumor showed MPNST with additional foci of epithelioid, rhabdomyoblastic, osteogenic and lipogenic differentiation. Although the capacity of MPNST to undergo epithelioid, rhabdomyoblastic, osteogenic and very rarely lipogenic differentiation is reported in literature, the occurrence of all these differentiation in one case has not been described in literature before. To the best of our knowledge, this is only the second MPNST case with lipomatous differentiation.

Mycobacterial culture of fine needle aspirate - a useful tool in diagnosing tuberculous lymphadenitis.

A prospective study was undertaken on suspected lymph node TB (LNTB) patients, to evaluate the diagnostic utility of mycobacterial culture of fine needle aspirate (FNA), in comparison with the cytological examination and acid fast staining. Eighty percent of 157 aspirates studied were positive by cytological examination; 18% by ZN smear and 45% were positive by culture. Twelve aspirates which were negative by cytological features yielded positive mycobacterial cultures; four out of these were from HIV positive patients. Our observations suggest that supplementing FNA cytology with mycobacterial culture would increase the sensitivity of diagnosing LNTB; in addition to giving a highly specific diagnosis.

A study of mycobacterial species causing lymphadenitis.

This prospective study evaluated the non-tuberculous mycobacteria (NTM) cases of lymphadenitis. A total of 76 isolates of mycobacteria were obtained from 200 lymph node aspirates suspected of tuberculosis, 74 of which were Mycobacterium tuberculosis, one was Mycobacterium fortuitum and one Mycobacterium kansasii. These results highlight the importance of NTM in HIV-negative patients as a case of lymphadenitis, and indicates the re-emergence of NTM as potential lymph node pathogens in this part of the country. Further studies on a larger scale are needed to delineate the association between NTM infections in HIV positive and negative subjects.

Morbidity pattern of hydatid disease (cystic echinococcosis) and lack of its knowledge in patients attending Mamata General Hospital, Khammam, Andhra Pradesh.

There is hearsay that prevalence of hydatid disease in Khammam and Nalgonda districts of Andhra Pradesh is high. We report here a preliminary study conducted to determine the magnitude of the problem of hydatid disease and the morbidity associated with it in patients attending MGH, KMM, A.P. (rural hospital). Eleven cases were identified during the period from November 2005 to May 2006 (seven months). Pain in abdomen, mass per abdomen, loss of appetite, pregnancy complicated by cystic echinococcosis (CE), and jaundice were the main clinical symptoms and signs. Ultrasonography, detection and removal of the cysts on the operation table, microscopic examination of the aspirated hydatid fluid were confirmatory. Ziehl-Neelsen stain of the aspirated fluid revealed acid-fast scolices. Interrogation of the patients and their family members (50) revealed that there was a total lack of knowledge of dog-tapeworm-caused infection in humans. They knew 'rabies' as the only disease man gets from dogs, and tapeworms are from pork and beef.

Road traffic accident (RTA) as major killer: a report on medico-legal autopsies in Bharatpur hospital.

Road traffic accidents are the major cause of medico-legal deaths. In present study out of 229 autopsy cases in one year, 110 deaths were due to road traffic accident (R.T.A.) which shows the magnitude of the problem in Chitwan and adjacent districts in Nepal.

Symbiotic characterization of isoleucine+valine and leucine auxotrophs of Sinorhizobium meliloti.

Ten isoleucine+valine and three leucine auxotrophs of Sinorhizobium meliloti Rmd201 were obtained by random mutagenesis with transposon Tn5 followed by screening of Tn5 derivatives on minimal medium supplemented with modified Holliday pools. Based on intermediate feeding, intermediate accumulation and cross-feeding studies, isoleucine+valine and leucine auxotrophs were designated as ilvB/ilvG, ilvC and ilvD, and leuC/leuD and leuB mutants, respectively. Symbiotic properties of all ilvD mutants with alfalfa plants were similar to those of the parental strain. The ilvB/ilvG and ilvC mutants were Nod-. Inoculation of alfalfa plants with ilvB/ilvG mutant did not result in root hair curling and infection thread formation. The ilvC mutants were capable of curling root hairs but did not induce infection thread formation. All leucine auxotrophs were Nod+ Fix-. Supplementation of leucine to the plant nutrient medium did not restore symbiotic effectiveness to the auxotrophs. Histological studies revealed that the nodules induced by the leucine auxotrophs did not develop fully like those induced by the parental strain. The nodules induced by leuB mutants were structurally more advanced than the leuC/leuD mutant induced nodules. These results indicate that ilvB/ilvG, ilvC and one or two leu genes of S. meliloti may have a role in symbiosis. The position of ilv genes on the chromosomal map of S. meliloti was found to be near ade-15 marker.

Symbiotic characteristics of cysteine and methionine auxotrophs of Sinorhizobium meliloti.

Twenty one cysteine and 13 methionine auxotrophs of Sinorhizobium meliloti Rmd201 were obtained by random mutagenesis with transposon Tn5. The cysteine auxotrophs were sulfite reductase mutants and each of these auxotrophs had a mutation in cysI/cysJ gene. The methionine auxotrophs were metA/metZ, metE and metF mutants. One hundred per cent co-transfer of Tn5-induced kanamycin resistance and auxotrophy from each Tn5-induced auxotrophic mutant indicated that each mutant cell most likely had a single Tn5 insertion. However, the presence of more than one Tn5 insertions in the auxotrophs used in our study cannot be ruled out. All cysteine and methionine auxotrophs induced nodules on alfalfa plants. The nodules induced by cysteine auxotrophs were fully effective like those of the parental strain-induced nodules, whereas the nodules induced by methionine auxotrophs were completely ineffective. The supplementation of methionine to the plant nutrient medium completely restored symbiotic effectiveness to the methionine auxotrophs. These results indicated that the alfalfa host provides cysteine but not methionine to rhizobia during symbiosis. Histological studies showed that the defective symbiosis of methionine auxotrophs with alfalfa plants was due to reduced number of infected nodule cells and incomplete transformation of bacteroids.

Ultrastructural studies on nodules induced by pyrimidine auxotrophs of Sinorhizobium meliloti.

Twenty three pyrimidine auxotrophs of Sinorhizobium meliloti Rmd201 were generated by random mutagenesis with transposon Tn5. On the basis of biochemical characters these auxotrophic mutants were classified into car, pyrC and pyrE/pyrF categories. All auxotrophs induced white nodules which were ineffective in nitrogen fixation. Light and electron microscopic studies revealed that the nodules induced by pyrC mutants were more developed than the nodules of car mutants. Similarly the nodules induced by pyrE/pyrF mutants had more advanced structural features than the nodules of pyrC mutants. The nodule development in case of pyrE/pyrF mutants was not to the extent observed in the parental strain. These results indicated that some of the intermediates and/or enzymes of pyrimidine biosynthetic pathway of S. meliloti play a key role in bacteroidal transformation and nodule development.

Isolation and symbiotic characterization of aromatic amino acid auxotrophs of Sinorhizobium meliloti.

Ten aromatic amino acid auxotrophs of Sinorhizobium meliloti (previously called Rhizobium meliloti) Rmd201 were generated by random mutagenesis with transposon Tn5 and their symbiotic properties were studied. Normal symbiotic activity, as indicated by morphological features, was observed in the tryptophan synthase mutants and the lone tyrosine mutant. The trpE and aro mutants fixed trace amounts of nitrogen whereas the phe mutant was completely ineffective in nitrogen fixation. Histology of the nodules induced by trpE and aro mutants exhibited striking similarities. Each of these nodules contained an extended infection zone and a poorly developed nitrogen fixation zone. Transmission electron microscopic studies revealed that the bacteroids in the extended infection zone of these nodules did not show maturation tendency. A leaky mutant, which has a mutation in trpC, trpD, or trpF gene, was partially effective in nitrogen fixation. The histology of the nodules induced by this strain was like that of the nodules induced by the parental strain but the inoculated plants were stunted. These studies demonstrated the involvement of anthranilic acid and at least one more intermediate of tryptophan biosynthetic pathway in bacteroidal maturation and nitrogen fixation in S. meliloti. The alfalfa plant host seems to provide tryptophan and tyrosine but not phenylalanine to bacteroids in nodules.

Hemostatic alterations in non-cirrhotic portal fibrosis, extrahepatic portal venous obstruction and Budd-Chiari syndrome.

We studied the coagulation function in ten patients each with non-cirrhotic portal fibrosis (NCPF), extrahepatic portal venous obstruction (EHPO) and Budd-Chiari syndrome (BCS), conditions where venous thrombosis in the hepatic vasculature is a common denominator. Prothrombin time, partial thromboplastin time with kaolin (PTTK) and thrombin time were normal in patients with NCPF and EHPO. However, in BCS the PTTK was prolonged, with a mean test/control ratio of 1.68 +/- 0.11. Fibrin degradation products were absent in all patients. Platelet aggregation tests showed hypoaggregability in all patients with NCPF. They were normal in patients with EHPO. However, two of ten BCS patients showed hyperaggregability, coinciding with a recent onset of illness in one patient. In conclusion, coagulation abnormalities appear unlikely to be the cause of thrombosis in patients with NCPF and EHPO. Further studies are required to substantiate the findings of hyperaggregability of platelets in BCS.