Development and validation of RP-HPLC method for pitavastatin calcium in bulk and formulation using experimental design

The optimization of HPLC method involves several variables whose influence has been widely studied. However, inmost of the cases, only process variables are taken into account. In this work, the influence of mixture compositionon peak quality parameters of Pitavastatin calcium in bulk and tablet dosage form has been studied using a mixturesimplex design. A simplex centroid design with axial points in a pseudo-component representation was generated fromthe pure mixture components. Twelve ternary mixture mobile phases corresponding to augmented design points weretested to separate the drug in sample. The statistical analysis was performed to generate the polynomial equation foreach response. The desirability approach was used to determine the optimal mobile phase composition. Furthermore,the method was validated as per the ICH guidelines using specificity, linearity, accuracy, precision, sensitivity, systemsuitability, and robustness. The results of experimental design were statistically tested for full and in portion to getbest fitted model which accurately describe changes in the proportion of these solvents in the mobile phase close to theregion of optimal peak quality. The method demonstrated optimum chromatographic separation with isocratic elutionof the mobile phase containing a mixture of acetonitrile-water (pH 3.0)-tetrahydrofuran (43:55:02, v/v/v) with a flowrate at 1.0 ml/minute. Design of experiment optimization strategy is a powerful tool to acquire the maximum qualitydata while performing minimum number of experiments. The mobile phase composition was successfully optimizedusing simplex centroid mixture design with desirability approach. Additionally, developed method can be appliedfor routine quantitative analysis of Pitavastatin calcium in bulk and tablet dosage form as it was found to be simple,sensitive, and robust.

Clinical and microbiological profile of febrile neutropenia in solid tumors and hematological malignancies at a tertiary cancer care center in South India.

BACKGROUND: Febrile neutropenia (FN) is a common but serious complication of chemotherapy in patients with solid tumors (ST) and hematological malignancies (HM). The epidemiology of FN keeps changing. OBJECTIVE: The objective was to study the epidemiology of FN in adult patients with ST and HM at Kidwai Memorial Institute of Oncology, Bangalore – A tertiary cancer care center. MATERIALS AND METHODS: Data of all episodes of FN that occurred during the period July 2011 to December 2011 were collected prospectively and analyzed. RESULTS: A total of 75 episodes of FN was observed during study period involving 55 patients. Febrile neutropenic episodes were more frequent in HM than in ST (57% vs. 43%). The rate of bloodstream infection was 14.7%. Gram‑negative organisms were the predominant isolates (56.25%). Overall mortality rate was 13.3%. Presence of medical co‑morbidity and positive culture predicted high mortality. Mortality rate did not differ significantly between HM and ST (14% vs. 12.5%; P = 1.0). Gram‑positive bacteremia was associated with greater mortality than Gram‑negative bacteremia (P = 0.02). CONCLUSION: Empiric antibiotic treatment for FN should be tailored to the locally prevalent pathogens and their susceptibility patterns.

A study of correlation between iron deficiency anaemia and serum lipid profile in Indian adults in BRIMS, Bidar.

Both iron deficiency anemia and dyslipidaemia are widely prevalent public health problems, especially in the Indian population. Some link has been suggested between the two potentially morbid conditions but a sufficient Indian study could not be found in this regard. Methods: This study was planned to find the changes in serum lipid profile in adult Indian patients with iron deficiency anemia and the effect of oral iron therapy on them. 100 iron deficiency anemia and 70 age and sex matched healthy controls, in the age group 18-35 years were investigated for any possible changes in serum lipid profile i.e., triglycerides, total cholesterol, high density lipoprotein cholesterol, very low density lipoprotein cholesterol. The patients were followed up after 3 months of oral iron therapy. Results: The results are shown as mean± standard deviation. Triglycerides and very low density lipoprotein cholesterol levels were found to be significantly (P <0.001) elevated in the iron deficiency anemia group (151.87 ± 48.06 mg/dl and 30.40 ± 9.71 mg/dl) as compared to controls (109.99 ± 30.81 mg/dl and 21.96 ± 6.69 mg/dl), whereas level of low density lipoprotein cholesterol were found to be significantly (P = 0.02) lower in patients (90.96 ± 41.55 mg/dl) as compared to controls (105.24 ± 26.45 mg/dl). However, after treatment (in 43 patients) there was significant (P <0.001) reduction in the levels of triglycerides and very low density lipoprotein cholesterol (111.56 ± 26.87 mg/dl and 22.30 ± 5.36 mg/dl) when compared to their pretreatment level (154.70 ± 53.89 mg/dl and 30.93 ± 10.84 mg/dl), whereas low density lipoprotein cholesterol levels did not show any significant change. Conclusion: These findings indicate that iron deficiency anemia in Indian adults is attended by abnormal serum lipid profile, which responds significantly to iron therapy.

Anatomy and histophysiological changes in pituitary of calotes versicolor during beeding and nnbreeding sasons / Anatomía y cmbios hstofisiológicos en la glándula pituitaria de la calotes versicolor durante las temporadas de crianza y la no reproductiva

Calotes versicolor, collected during peak breeding and nonbreeding season were used for the study of anatomy and histophysiology of pituitary gland. In C. versicolor the pituitary gland is small compact mass attached to the hypothalamus by a delicate stalk. It is of two basic units, adenohypophysis and neurohypophysis. The adenohypophysis includes pars distalis (PD) and pars intermedia (PI). The pars tuberalis (PT) which is usually reduced in reptiles, is absent in C. versicolor. The neurohypophysis is composed of median eminence (MI) and neural lobe or pars nervosa (PN). The anterior PD is rich acidophils than the posterior PD during breeding season. These cells are differentiated as A1 and A2 cells. The A1 cells are erythrosinophilic and stained purple or red and A2 cells stained yellow with orange G. The identified A1 cells are regarded as lactophores and A2 cells are regarded as somatotropes. The secretion of these cells and cell number is reduced during nonbreeding season. The basophils of PD are PAS +, AF + and aniline blue + during breeding season. These cells are classified as B1 and B2 cells depending on their size and location. The large B2 cells are present in cords and B1 cells are located closely to these parts. The B1 and B2 cells are regarded as thyrotrophs and gonadotrophs respectively. During nonbreeding these cells show negative response to the above stain. The PI is large and labulated. The cells are AF+, erythrosine and secrete highly granular material during breeding season. During nonbreeding season they become AF-, erythrosine- and less secretory. The secretion present in neurosecretory cells of the neural lobe appears as large granules and they are AF+. These cells take deep blue stain indicating rich neurosecretory material during both breeding and nonbreeding season. The median eminence (MI) is located at the anterior part of neurohypophysis and sends portal capillaries to adenohypophysis.

Calotes versicolor, recolectadoss durante el peak de la temporada no reproductiva y la reproductiva fueron utilizados para el estudio de la anatomía e histofisiología de la glándula pituitaria. En C. versicolor la glándula pituitaria es una masa compacta y pequeña unida al hipotálamo por un tallo delgado. Se trata de dos unidades básicas, adenohipófisis y neurohipófisis. La adenohipófisis incluye la pars distalis (PD) y la pars intermedia (PI). La pars tuberalis (PT), usualmente reducida en reptiles, está ausente en C. versicolor. La neurohipófisis está compuesta por una eminencia media (EM) y el lóbulo neural o pars nerviosa (PN). La PD anterior es más rica en acidófilos que la PD posterior durante la temporada de reproducción. Estas células se diferencian como células A1 y A2. Las células A1 son eritrosinofílicas y se tiñen de color púrpura o rojo y A2 son teñidas de amarillo con anaranjado G. Las células identificadas A1 se consideran lactóforas y las células A2 se consideran somatotropas. La secreción de estas células y el número de células se reduce durante la temporada no reproductiva. Las basófilas de la PD son PAS+, AF+ y azul anilina + durante la temporada de cría. Estas células se clasifican como las células B1 y B2 en función de su tamaño y ubicación. Las grandes células B2 están presentes en cordones y B1 se encuentran localizadas cerca de estas partes. Las células B2 y B1 se consideran tirotrofos y gonadotrofos respectivamente. Durante la temporada no reproductiva de estas células muestran una respuesta negativa a la tinción anterior. La PI es grande y labulada. Las células son AF+, eritrosina+ y secretan material altamente granular, durante la temporada de cría. Durante la temporada no reproductiva se convierten en AF-, eritrosina- y de menos secreción. La secresión presente en las células neurosecretoras del lóbulo neural aparece en forma de gránulos grandes y es AF+. Estas células toman una profunda tinción azul indicando el rico material..

Effect of ethanol extract of whole plant of Trichosanthes cucumerina var. cucumerina L. on gonadotropins, ovarian follicular kinetics and estrous cycle for screening of antifertility activity in Albino Rats / Efectos del etanol extraído de la planta completa de Trichosanthes cucumerina var. cucumerina L. sobre gonadotropinas, cinética folicular ovárica y ciclo para screening de actividad antifertilidad en Ratas Albinas

Ethanol extract of whole plant of Trichosanthes cucumerina L. var. cucumerina was evaluated for antiovulatory activity in adult rats. The ethanol extract at the doses 200 and 400mg/kg body weight (orally) affected the normal estrous cycle showing a significant increase in estrus and metestrus phases and decrease in diestras and proestrus phases. The extract also significantly reduced the number of healthy follicles (Class I-Class VI) and corpora lutea and increased the number of regressing follicles (Stage IA, Stage IB, Stage IIA, and Stage IIB). The protein and glycogen content in the ovaries were significantly reduced in treated rats. The cholesterol level was significantly increased, whereas, the enzyme activities like 3b-HSD and 17b-HSD were significantly inhibited in the ovary of treated rats. Serum FSH and LH levels were significantly reduced in the treated groups were measured by RIA. In acute toxicity test, neither mortality nor change in the behavior or any other physiological activities in mice were observed in the treated groups. In chronic toxicity studies, no mortality was recorded and there were no significant differences in the body and organ weights were observed between controls and treated rats. Hematological analysis showed no significant differences in any of the parameters examined (RBC, WBC count and Hemoglobin estimation). These observations showed the antiovulatory activity of ethanol extract of whole plant of Trichosanthes cucumerina L. var. cucumerina in female albino rats.

El extracto de etanol de toda la planta de Trichosanthes cucumerina var. cucumerina (L.) se evaluó en cuanto a su actividad antiovulatoria en ratas adultas. El extracto de etanol en dosis de 200 y 400mg/kg de peso corporal (oral) afectó el ciclo normal estral, mostrando un aumentó significativo en las fases de estro y metaestro y la disminución de las fases de diestro y proestro. El extracto también redujo significativamente el número de folículos sanos (Clase I=Clase VI) y cuerpo lúteo y aumentó el número de folículos en regresión (etapa I, etapa IB, etapa II y etapa IIB). La proteína y el contenido de glucógeno en los ovarios se redujeron significativamente en las ratas tratadas. El nivel de colesterol aumentó significativamente, mientras que, actividad de las enzimas 3b-HSD y 17b-HSD se inhibió significativamente en el ovario de ratas tratadas. FSH sérico y los niveles de LH se redujeron significativamente en los grupos tratados y medidos por RÍA. En la prueba de toxicidad aguda, no hubo mortalidad ni cambio en el comportamiento fisiológico o de cualquier otra actividad en los grupos tratados de ratas. En estudios de toxicidad crónica, no se registró mortalidad y no hubo diferencias significativas en el peso corporal o el peso de los órganos entre los controles y las ratas tratadas. Los análisis hematológicos no mostraron diferencias significativas en ninguno de los parámetros examinados (eritrocitos, recuento de glóbulos blancos y estimación de hemoglobina). Estas observaciones mostraron la actividad antiovulatoria del extracto de etanol de toda la planta de Trichosanthes cucumerina var. cucumerina en ratas albinas hembras.

Effect of FSH and LH on testis during nonbreeding season in Calotes versicolor (Daud.).

Histological and biochemical studies carried out on the male reproductive organs of the Indian garden lizard, Calotes versicolor after the treatment with pituitary gonadotrophins (FSH and LH), showed a significant increase in the weight, protein content and diameter of testis, but decrease in its cholesterol. The spermatogonia, spermatocytes and spermatids increased significantly in the germinal epithelium of seminiferous tubule, and spermatozoa appeared in its lumen. The Leydig and Sertoli cells were hypertrophied with increase in their nuclear diameter. The epidymal weight, diameter and protein content also increased after gonadotrophins treatment. There was a significant decrease in the testicular cholesterol indicating the utilization of cholesterol for steroid hormone synthesis. The combined gonadotrophin (FSH + LH) treatment was more effective than the individual gonadotrophin treatment.

Fusarium solani fungemia in a patient with acute lymphoblastic leukemia.

A patient with ALL on anticancer chemotherapy developed fever which was later attributed to be due to Fusarium fungemia. The details of the case & a review of literature follows.

Bacterial infection and endotoxin in female reproductive tract in rats: correlation with the developmental status of preimplantation embryos.

During mammalian preimplantation development, a substantial numbers of embryos are believed to be lost for reasons that are unclear. Using female rats, we investigated whether the developmental status of embryos is influenced by bacterial infection and endotoxin in the reproductive tract. From the vagina of cycling rats (n = 11), 21 bacterial isolates were identified; they were Streptococcus faecalis (S. faecalis; 38%), Escherichia coli (E. coli; 19%), Acinetobactor calcoaceticus (A. calcoaceticus; 14%), and coagulase negative staphylococcus (14%), Micrococcus sp. (5%), Bacillus subtilis (B. subtilis; 5%) and Proteus vulgaris (P. vulgaris; 5%). From the vagina of day 4 pregnant rats (n = 12), 26 isolates were identified; they were S. faecalis (23%), A. calcoaceticus (23%), E. coli (15%), Micrococcus sp. (15%), B. subtilis (8%), P. vulgaris (4%), Staphylococcus aureus (4%), beta-hemolytic streptococcus (4%) and Pseudomonas aeruginosa (4%). Gram negative bacteria found in the vagina of cycling and day 4 pregnant rats were 38% and 46%, respectively. In both, bacterial load was 10(3)-10(5) colony forming units and there was no association with the abnormality of the recovered embryos. However, in two day 4 pregnant animals, pathogenic bacteria (Staphylococcus aureus and beta-hemolytic streptococcus) were isolated and embryos recovered from them were degenerated and deformed. The vagina of day 9 pregnant animals (n = 7) were, however, sterile. Consistently, in all animals, the upper reproductive tract (uterus and oviduct) was devoid of any bacteria and no anaerobic bacteria were isolated from any part of the tract. The levels of endotoxin in the vagina of cycling and day 4 pregnant rats were 1.35 +/- 0.1 and 1.17 +/- 0.1 endotoxin units (EU), respectively. It was undetectable in the oviduct and uterus of all animals (n = 5) except one which showed high levels of endotoxin in uterus (4.5 EU) and oviduct (2.2 EU) and the animal also produced degenerated and deformed embryos. These results indicate that common bacterial flora of vagina may not affect embryo development and the presence of pathogenic bacteria in the vagina and/or endotoxin in reproductive tract could be detrimental to viability of gametes and preimplantation embryos in rats.