Azadirachta indica (Meliaceae) and its derived products have shown a variety of insecticidal properties. The present paper discusses the larvicidal activity of neem-based biopesticide for the control of mosquitoes.Mosquitoes transmit serious human diseases, causing millions of deaths every year. Use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of botanical origin have been reported as useful for control of mosquitoes. rd and early 4th instar larvae of different genera of mosquitoes. The larvae were exposed to different concentrations (0.5–5.0 ppm) of the formulation along with untreated control. Larvicidal activity of the formulation was also evaluated in field against Anopheles, Culex, and Aedes mosquitoes. The formulation was diluted with equal volumes of water and applied @ 140 mg a.i./m2 to different mosquito breeding sites with the help of pre calibrated knapsack sprayer. Larval density was determined at pre and post application of the formulation using a standard dipper.Larvicidal efficacy of an emulsified concentrate of neem oil formulation  developed by BMR & Company, Pune, India, was evaluated against late 350) of the formulation against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was found to be 1.6, 1.8 and 1.7 ppm respectively. LC50 values of the formulation stored at 26°C, 40°C and 45°C for 48 hours against Ae. aegypti were 1.7, 1.7, 1.8 ppm while LC90 values were 3.7, 3.7 and 3.8 ppm respectively. Further no significant difference in LC50 and LC90 values of the formulation was observed against Ae. aegypti during 18 months storage period at room temperature. An application of the formulation at the rate of 140 mg a.i./m2 in different breeding sites under natural field conditions provided 98.1% reduction of Anopheles larvae on day 1; thereafter 100% reduction was recorded up to week 1 and more than 80% reduction up to week 3, while percent reduction against Culex larvae was 95.5% on day 1, and thereafter 80% reduction was achieved up to week 3. The formulation also showed 95.1% and, 99.7% reduction of Aedes larvae on day 1 and day 2 respectively; thereafter 100% larval control was observed up to day 7.Median lethal concentration (LCThe neem oil formulation was found effective in controlling mosquito larvae in different breeding sites under natural field conditions. As neem trees are widely distributed in India, their formulations may prove to be an effective and eco-friendly larvicide, which could be used as an alternative for malaria control. Mosquitoes transmit serious human diseases like malaria, filariasis, Japanese encephalitis, dengue haemorrhagic fever and yellow fever causing millions of deaths every year . ExtensiAzadirachta indica) native of India, belonging to family Meliaceae are fast growing evergreen trees ranging in height from 12 – 24 m. They are widespread in tropical and subtropical regions of the world, including semi-arid and wet- tropical regions [Neem trees, , sorbitan dioleate (surfactant) and epichlorohydrin (used as a stabiliser to protect the degradation of the formulation under exposure to sun light.), developed by BMR & Company, Pune, India was evaluated against late 3rd and early 4th instar larvae of Anopheles stephensi, a primary vector of urban malaria,Culex quinquefasciatus a common vector of filariasis, and Aedes aegypti a common vector of dengue, dengue haemorrhagic fever and yellow fever. The larvae were obtained from laboratory-established colony as described earlier [50)/90% lethal concentration (LC90) of the formulation.Larvicidal bioassay of the formulation was performed on late 3 earlier . Twenty- earlier  and Log Ae. aegypti at different concentration (0.5–5.0 ppm) as per the method reported above. Five replicates of each concentration were run under the same microclimatic conditions along with untreated control. A total of 100 larvae of Ae. aegypti were exposed against each concentration. Further bioassay test of the neem oil formulation of different concentrations (0.5–5.0 ppm) stored at room temperature (26°C ± 2°C) was evaluated against Ae. aegypti larvae at three months interval period for 18 months. Three replicates of each concentration were carried out along with control.Larvicidal bioassay of the neem oil-based formulation stored at 26°C, 40°C and 45°C for 48 hrs was evaluated against rd and early 4th instar larvae of different genera of mosquitoes under natural field conditions.Field evaluation of larvicidal activity of the neem oil formulation was carried out in two districts of Uttar Pradesh viz. Mathura and Kanpur and district Hardwar of the state of Uttarakhand. Initially a survey was carried out to ascertain the suitability of different breeding habitats of target species for the trial. Larvicidal efficacy was carried out against late 3a.i./m2) of the formulation was determined on the basis of the results of laboratory evaluation carried out against Cx. quinquefasciatus and Ae. aegypti [2 surface area of breeding habitats through precalibrated knapsack sprayer.Larval density was determined using standard dipper (300 ml capacity with 9 cm diameter) method. Treatment dose  of the neem oil formulation against An. stephensi, Cx. quinquefasciatus and Ae, aegypti are given in Table 50 values of the formulation were 1.6, 1.8 and 1.7 ppm while LC90 were 3.4, 3.5 and 3.7 ppm against An. stephensi, Cx. quinquefasciatus and Ae. aegypti respectively. Results of stability test of the neem oil based formulation stored at different temperatures against Ae. aegypti are given in Table 50 values of the biopesticide stored at 26°C, 40°C and 45°C for 48 hours were 1.7, 1.7, 1.8 ppm while LC90 values recorded were 3.7, 3.7 and 3.8 ppm respectively. Mean LC50 and LC90 of the formulation during 18 months storage period at room temperature against Aedes aegypti is shown in Figure 50 and LC90 value of the formulation was observed.Mean LCAedes larvae. Larvae collected from these breeding sites were identified as Ae. aegypti and Ae. albopictus. A survey of different breeding sites such as tanks, pits, drains were also carried at Indian Oil Corporation, Mathura and Bharat Heavy Electricals Limited, Hardwar for Culex and Anopheles breeding. Culex and Anopheles larvae collected from these sites were identified as Cx. quinquefasciatus, An. culicifacies and An. subpictus respectively.Before start of the study, a preliminary survey of various breeding sites of mosquitoes was carried out. In all 73 breeding sites (factory scraps) were surveyed inside the Ordnance factory, Kanpur, out of which 67 sites were found positive (91.8%) for Cx. quinquefasciatus and anophelines in different breeding habitats are given in Table Culex larvae was 95.9, 90.2, 87.2 on days 1, 2, 3 respectively of post application, while more than 70% reduction was observed up to week 3. In tanks 91.6% – 92.4% reduction of Culex larvae was observed up to day 7 of post application thereafter 80.7% reduction was noted up to week 3, while in drains there was more than 90% larval control up to day 7 and remained above 75% up to week 3. An effective control of late instars anopheline larvae in tanks was observed with 98.2% reduction on day 1, followed by 100% reduction up to day 7 and more than 75% reduction till week 3. In pits, 96.2% control was recorded on day 1, 100% up to day 7 and more than 75% reduction up to week 3. The mean percent reduction of Culex larvae was 89.9–95.5% up to day 7 followed by 79.7–85.7% up to week 3, while for Anopheles larvae mean percent reduction was 90.4–100% up to day 7 followed by 83.8–90.4% up to week 3 , an emulsifier (5%) and 63% iso propanol (solvent) was investigated for its larvicidal activities against An. gambiae [50 value of 11 ppm and also reported to possess insect growth regulators. Gianotti and co workers [An. gambiae at the rate of 10 gm/m2 of pool surface area for effective larval control. Azadirachtin acts as anti-ecdysteroid and kills larvae by growth inhibition effect [Anopheles, Culex and Aedes larvae was observed up to three weeks of post application.Dhar et al  demonstr gambiae . It was  workers  used pown effect . In the Neem-based biopesticides and neem extracts have a wide range of effects against insect pests including repellence, feeding, toxicity, sterility and growth regulator activity and are relatively safe towards non- target biota with only minimal risk of direct adverse effects on aquatic biota from contamination of water bodies ,22. AlleBacillus thuringiensis is a matter of concern for operational use as larvicides. Although the present formulation may be more costly than other larvicidal agents, such as temephos and B. thuringiensis, it has the advantage of being eco-friendly, effective and ability to prevent the development of pest resistance.Control of mosquito larvae becomes a very pertinent issue in controlling the rapid replication of mosquitoes in management of vector- borne diseases. In the present study, neem oil formulation showed promising larvicidal activity against important vectors of malaria, filaria, dengue, dengue haemorrhagic fever, yellow fever and chikungunya. Development of resistance in temephos and The neem oil formulation was found effective in controlling mosquito larvae in different breeding sites under natural field conditions. Neem oil formulations are relative less toxic, eco-friendly and insects are unable to develop resistance and may be used as an alternative to other pesticides for control of vector- borne diseases.The authors declare that they have no competing interests.VD: designed the study protocols, directed the larvicidal research. ACP: designed and performed the larvicidal trials collected and analysed the data. KR: Editing the manuscriptAG: Performed larvicidal trials drafted the manuscript TS: Field trials of the larvicide. APD; Coordination and organizational help to conduct the study. All authors read and approved the final manuscript.


The macrophage content of cell suspensions from naturally occurring mouse tumours has been assessed by the Fc-mediated phagocytosis assay, and the results compared with the individual tumour's capacity for spontaneous metastasis and with its pulmonary colonization potential after i.v. inoculation. It was found that these tumours differ in their properties from the transplantable fibrosarcomas studied previously by other investigators, in that the macrophage content of all the tumours was uniformly low, ranging from 2 to 9% (mean 4.2 +/- 1.8%) and there was no inverse correlation with frequency of spontaneous metastasis, which was low. When the tumours were inoculated i.v. there was also no correlation with colony-forming capability, which varied greatly between tumours. Lung secondary deposits contained 1.7-6% macrophages (mean 4.4 +/- 0.6%) with a lower phagocytic activity for antibody-coated red cells than in the primary tumour.


The release of ondansetron hydrochloride from the gel matrix showed diffusion- controlled.Ondansetron is a serotonin receptor antagonist used in the management of nausea and vomiting that is associated with cancer chemotherapy. There is a need for intranasal delivery due to poor bioavailability of drug because of first pass effect. The objective of this study was to develop an intranasal delivery system of ondansetron hydrochloride using thermo-sensitive polymer PF127 and mucoadhesive polymer hydroxypropylcellulose. Due to increase in bioadhesive polymer concentration, there was increase in bioadhesion strength, at the same time there was decrease in the spredability. An  There is a need for intranasal delivery due to poor bioavailability of drug because of first pass effect[Ondansetron hydrochloride is a serotonin (5HTss effect. The objLutrol F127 (PF127 and hydroxypropylcellulose (Klucel LF) was procured from Signet Chemicals Mumbai. Ondansetron hydrochloride was received as a gift sample from Dr. Reddy's Lab Hyderabad.et al.[Nasal formulations consisting of aqueous gels of PF127 containing 18% (w/v) of polymer were prepared using the method described by Schmolka. Rheologet al..A glass cup with a cross-sectional area of 7.5 cmnd and adhesive tape, and inverted under the surface of 500 ml of simulated nasal fluid of pH 5.5 at 35±0.5° in USP XXIII Type I Dissolution Test Apparatus with a speed of 50 rpm. Five millilitres of aliquots were withdrawn at specified time intervals and immediately replaced with fresh dissolution medium. The dru2 but maximum spredability and 88.2% drug release after 8 h. In all formulations 0.2% w/v bioadhesive polymer showed maximum spredability while 0.7% w/v showed minimum spredability (All gels were glassy clear in appearance. Gel pH was in the range of 5.3 to 5.6 which was in the range of pH at the absorption site (4.5-6.5). In the preliminary studies, the minimum concentration of PF127 that formed gel below 34° was found to be 18% wt/vol. The plain formulation (F1) without bioadhesive polymer has minimum bioadhesion 529.2 dyne/cmdability . There wdability , fig. 1.dability .in situ gels can be formulated of ondansetron hydrochloride using PF127 and bioadhesive polymer.Various solubilizers were tried during preformulation study. Propylene glycol and Transcutol were selected as solubilizers for further study. Poloxamer 407 gives gelation temperature in nasal temperature range 28° to 35° in 18% w/v concentration. Addition of bioadhesive polymer from 0.2% to 0.7% further lowered the gelation temperature from 34° to 29°. Gelation temperatures obtained using two different methods  did not vary more than ±1°. The gelation temperature lowering effect of bioadhesive polymer might have caused in part by the increased viscosity after dissolution of mucosadhesive polymer. The pH values of all formulations were found in the range of 5.3 to 5.6. Formulation should posses' mild acidic pH for activation of lysozyme . The pKa of ondansetron hydrochloride is 7.4; so drug present in solubilised form in pH range 5 to 6.5 (ondansetron hydrochloride precipitate above 6.8). It was observed that the concentration of bioadhesive polymer increased from 0.2% to 0.7% showed retardation of ondansetron hydrochloride release, probable mechanism for such retardation of release may be reduction in number and dimensions of the channels in gel structure by increased viscosity of the formulation. From the study it can be concluded that the nasal


Chemoradiation for cervical cancer patients is associated with considerable gastrointestinal toxicity. Intensity-modulated radiotherapy (IMRT) has demonstrated superiority in terms of target coverage and normal tissue sparing in comparison to conventional 3D planning in gynaecological malignancies. Whether IMRT in prone (PP) or supine position (SP) might be beneficial for cervical cancer patients remains partially unanswered.mean 50.4 Gy) were optimized in terms of PTV coverage (1st priority) and small bowel sparing (2nd priority). A comparison of DVH parameters for PTV, small bowel, bladder, and rectum was performed.10 patients on FIGO stage IB-III cervical cancer, 6 patients for definitive and 4 patients for adjuvant external beam pelvic RT, were planned in PP and SP using a 7-field IMRT technique. IMRT plans for PP and SP . PTV, rectum and bladder volumes were comparable for PP and SP. Significantly larger volumes of small bowel were found in PP . PP decreased the volume of small bowel at 20-50.4 Gy (p < 0.05) and increased the rectum volumes covered by doses from 10-40 Gy (p < 0.01), the V50.4 was < 5% in both treatment positions. Bladder sparing was significant better at 50.4 Gy (p = 0.03) for PP.In this dosimetric study, we demonstrated that pelvic IMRT in prone position for patients with cervical cancer seems to be beneficial in reducing small bowel volume at doses ≥20 Gy while providing similar target coverage and target conformity. The use of frequent image guidance with KV (kilovolt) or MV (megavolt) computertomography can reduce set-up deviations, and treatment in prone position can be done with a higher set-up accuracy. Clinical outcome studies are needed to affirm lower toxicity. Chemoradiation is the treatment of choice in locally advanced, lymph node positive and/or high-risk cervical cancer patients -9. The t10 patients with histological confirmed cervical cancer on FIGO stage IB1, IB2, IIB and IIIB were selected for this study. 6 patients were treated with definitive chemoradiation . All of these patients underwent transperitoneal laparoscopic pelvic and paraaortic lymphadenectomy as described previously . Pelvic ® from GE Healthcare, General Electric Company, NYSE; GE), from the diaphragm to the trochanter minor with a slice thickness of 3.75 mm. were performed. The CT was performed with a belly board in prone position (PP), and with a head rest, a knee and ankle fixation in the supine position (SP). No instructions for bladder or rectum filling were given. The CT scans were acquired during a period when both scans were routinely carried out for patients with a high risk for paraaortic metastases, in order to switch the treatment technique easily if paraaortic metastases were histologically confirmed. Intravenous contrast mediaum (Xenetix 350®) and a vaginal tampon soaked in contrast medium were used to aid the delineation; oral contrast media was not used obligatorally.For each patient, two consecutive treatment planning CT scans  50 and 62, target volumes and organs at risk were delineated in all axial CT slices ,38. For mean). Treatment was performed on a linear accelerator  with 20 MV photon beams. Planning parameters for the PTV were set to minimize the amount receiving < 95% of the prescribed dose and the amount receiving > 105% (52.9 Gy) of the prescribed dose. The second highest priority was given to the sparing of small bowel. Inverse planning input parameters are shown in table The Eclipse Planning Software  was used to generate IMRT plans for SP and PP. The prescribed target dose was 5 × 1.8 Gy per week, to a total mean dose of 50.4 Gy . IMRT plans were generated based on a seven beam arrangement with beams at 45/90/115/180/245/280/320 degrees for SP and 0/40/80/115/235/270/320 degrees for PP, as is routinely applied at our clinic. The sliding window technique was used. Although patients also received brachytherapy, for this analysis, only external beam irradiation has been taken into account.mean) for the PTV and the organs at risk were measured. Dosimetric parameters were compared by the non-parametric Wilcoxon exact signed rank test . Statistical significance was assumed for p ≤ 0.05.DVH parameters for the target volume and critical normal tissues were analysed, and the PTV95% (volume of PTV receiving 95% of the prescribed dose) and the D1% (highest dose delivered to 1% of the PTV) was calculated. The target conformity was calculated according to ICRU reports 50 and 62  ,38 and amean was 50.4 Gy in PP and SP, respectively. The mean volume of PTV95% was 97.0% (STD ± 1.2) for SP and 97.6% (STD ± 0.8) for PP. The mean D1% was 52.9 Gy (STD ± 0.2) and 52.8 Gy (STD ± 0.2) for SP and PP, respectively. The PTV that received 110% of the prescribed dose was < 0.01% in both groups. Conformity of IMRT plans for PP and SP IMRT gave similar results . All parameters did not reach statistical significance.The mean volume of the PTV was 1227.0 cc  ± 66.6) for SP and 1369.4 cc  for PP (p = 0.6). Dmean  showed a statistically significant difference  and for the PP group from 52.8 to 174.5 cc . In PP, a larger rectal volume was covered by the V10 to V40  and 43.1 Gy ± 2.8 (STD) (p = 0.7) for PP.The bladder volume displayed a highly individual range for both positions. In SP, the bladder volume ranged from 70.7 to 417.7 ml, with a mean value of 143.9 ml ± 98.3 (STD), and from 70.2 to 395.2 ml, with a mean of 137.0 ml ± 93.4 (STD) (p = 0.6) in PP. The dose-volume histogram for the bladder was significant better in PP at V50.4 (p = 0.03). At V10, V20, V30, V40 and V45, no significant differences were detected (Table mean was 25.9 Gy vs. 30.2 Gy for PP and SP (p = 0.049), respectively.The small bowel volume varied from 683.8 to 1825.9 cc  for SP and from 1193.4 to 2443.9 cc  for PP. Statistically significant larger volumes of small bowel were found in PP (p = 0.01). Figure Due to an overlap of target structures (lymph nodes) and organs at risk, there is a considerable rate of gastrointestinal and genitourinary morbidity in patients with cervical cancer undergoing pelvic irradiation . With a Few publications addressed the issue of IMRT and the patients' position ,48. In amean 50.4 Gy), but the sparing of small bowel at V50 was considerable better. The sliding window technique, the 20 MV photon beam and the different contouring of the small bowel might contribute to this volume reduction. Interestingly, Adli and colleagues reported on dose inhomogeneities of up to and over 130% of the prescribed dose for both IMRT techniques.The prescribed total dose in the study by Adli was 45 Gy, where the small bowel was defined as all individual loops. They observed an anterior movement of small bowel for patients in PP, as we did. A dosimetric benefit was found for PP irrespective of the IMRT techniques. They concluded that the magnitude of small bowel sparing did not merely depend on the prone treatment position, but on the specific IMRT technique used. In our study, we did observe a larger "small bowel" volume  in PP, evolving from an expansion of the peritoneal cavity not basically due to more small bowel loops but a widening of the space between the loops though the opening in the belly board and an cranial movement of small bowel in supine position. Our small bowel volume for the PP group at 45 Gy and 50.4 Gy was 13.7% and 2.5%, as compared to the data from Adli and colleagues who saw 12.5% and 10% (45 Gy) and 5% and 6.6% (50 Gy) for limited arc or extended arc, respectively . Our premean ranged form 45-50.4 Gy. All patients received 10 Gy HDR brachytherapy to the vaginal-cuff. Small bowel volumes  at 45 and 50 Gy were remarkably low in PP and SP . The IMRT treatment was well tolerated and only one single Grade 3 chronic gastrointestinal toxicity was reported. The authors found no correlation between gastrointestinal morbidity and dosimetric parameters among the opposed set-up positions after a median follow up of 19-20 months. They therefore concluded that a longer follow up is needed to detect any existing differences between the two approaches.No significant dosimetric benefit was seen between PP and SP in a more recently published study by Beriwal and colleagues . They anOur aim was to compare the best IMRT 7-field standard technique at our department for the prone position and the supine position. Many factors influence the magnitude of dose reduction to the organs at risk. One key issue is the target volume definition. Large amounts of small bowel are located in the upper pelvic region, around the upper iliac external and common iliac lymph nodes. Our target definition was comparable to those of other authors -29. FurtIn this dosimetric study, we demonstrated that pelvic IMRT in prone position on a belly board seems to be a useful tool to reduce small bowel volume at a dose ≥20 Gy whilst providing similar target coverage and target conformity for patients with cervical cancer. Despite this, new evidence supports a comparable set-up error for the prone and supine treatment positions. We recommend frequent onboard imaging with KV or MV CTs to assure optimal set-up accuracy. Nevertheless, only outcome studies will show if the dosimetric differences in small bowel sparing will lead to decreased acute and late gastrointestinal toxicity.The authors declare that they have no competing interests.CS did the collection and assembly of data, data analysis and interpretation, and the manuscript writing. SM carried out conception and design, and manuscript writing. YK and TM optimised the IMRT plans. MK and UJ carried out data interpretation. CK and AS helped with the collection of data and to draft the manuscript. VB helped with the interpretation of the data and to draft the manuscript. All authors read and approved the final manuscript.


The method was validated for linearity, range, accuracy, precision and specificity. Cilostazol was determined in tablet dosage form in range of 99.58-100.67% with 0.4600 standard deviation. Stress studies were conducted in acid and alkali hydrolysis with gradual increasing concentration. Cilostazol was found to be stable in various concentrations of acidic and alkaline.A simple, selective, precise and stability-indicating high-performance liquid-chromatographic method of analysis of cilostazol in pharmaceutical dosage form was developed and validated. The solvent system consisted of 10 mM phosphate buffer (pH 6.0):acetonitrile:methanol (20:40:40). Retention time of cilostazol in C18 column was 5.7 ± 0.1 min at the flow rate 1.3 ml/min. Cilostazol was detected at 248 nm at room temperature. The linear regression analysis data for the calibration plots showed good linear relationship with correlation coefficient value, r A number of pharmaceutical substances have ester or amide as functional groups which may undergo hydrolysis in solutions or in aqueous suspension. Hydrolytic reactions involve nucleophilic attack on labile bonds such as lactam, ester, amide, imine and so on, by water on the drug in the solution and it follows first order kinetics11H-tetrazol-5-y1)-butoxy]-3,4-dihydro-2(1H)– quinolinone  and solvents acetonitrile and methanol were HPLC grade from Merck Ltd., India. All other chemicals  were analytical grade from Merck Ltd., India. Cilostazol tablets  were procured from local market.For the RP-HPLC method development and hydrolytic degradation analysis of cilostazol instrument and separation variables are shown 2 = 0.9998. The method was validated according to ICH guidelinesAccurately weighed about 100 mg cilostazol was dissolved in 50 ml methanol (HPLC grade) and volume was made upto 100 ml with triple distilled water . The stock solution was diluted to obtain 0, 100, 200, 400, 800, 1600 and 3200 ng/ml solution of cilostazol. The dilutions were fillered through 0.45 μm membrane filter and injected. Chromatograms were plotted and repeated for six times. A calibration graph was plotted between the peak AUC vs concentration and regression equation was AUC= 43.45X+156.75 with correlation coefficient rTwenty tablets  were weighed and finely powdered. Powder equivalent to 100 mg of cilostazol was dissolved in 50 ml methanol (HPLC grade) and volume was made upto 100 ml with triple distilled water. The sample was sonicated for 15 min and filtered through Whatmann paper no. 41 . 10 milliliters of this stock was diluted up to 100 ml with 50% aqueous methanol  and then further diluted up to 100 ml obtain stock R (10 μg/ml). Aliquots of 10 μg/ml were diluted to obtain concentration of 800 ng/ml and filtered through 0.45 μm membrane filter. Samples were analysed and statistical calculations were carried out .Hydrolytic degradation in alkaline condition was carried out by dissolving accurately weighed 100 mg cilostazol in 50 ml methanol (HPLC grade) and volume was made upto 100 ml with 2 N NaOH. The solution was refluxed on water bath at 60º for 5 h. Aliquot of above solution was neutralized with 1 N HCl and diluted with diluent to obtain 800 ng/ml solutions. The sample solution was analysed and chromatogram was recorded. No degradation of cilostazol was found in 1 N NaOH at 60º after 5 h. Further, cilostazol was degraded in 2 N NaOH and 5 N NaOH and cilostazol was found to be stable.Hydrolytic degradation under acidic conditions was performed by dissolving 100 mg cilostazol in 50 ml acetonitrile (HPLC grade) and volume was made upto 100 ml with 2 N HCl. The solution was refluxed on water bath at 60º for 5 h. Aliquots of above solution was neutralized with 1 N NaOH and diluted with diluent to obtain 800 ng/ml solutions. The sample solution was analysed and chromatogram was recorded. No degradation of cilostazol was found in 1 N HCl at 60º after 5 h. Further, the solution was degraded in 2 N HCl and 5 N HCl and was found to be stable.Cilostazol tablets were analysed by validated RP-HPLC method and cilostazol was found in between 99.58-100.67% with relative standard deviation 0.0046. As cilostazol is insoluble in water and sodium hydroxide solution, cosolvent was required to perform alkali degradation of cilostazol. Acetonitrile as cosolvent was avoided for alkali degradation because it produces phase separation with 1 N or more concentrated NaOH solutionCilostazol is also insoluble in hydrochloric acid and methanol as cosolvent is avoided with high concentration of HCl due to presence of amide group in cilostazol. Methanol may react with amide group and produce significant experimental artifact components


No significant difference was found between 1st (46.0 ± 7.7) and 2nd (48.3 ± 7.8) year students. When comparing scores based on gender, a significant difference was present between males (41.2 ± 12.2) and females (46.1 ± 11.0).Significant changes were found between both 1st and 2nd year medical students in comparison with 4th year student's scores. These findings suggest that as student exposure to allopathic techniques and procedures increases during the last year of medical school, their attitudes toward CAM decrease. Females were also significantly more likely to have stronger positive attitudes toward CAM than males, though both genders represented an overall positive attitude toward CAM.CHBQ scores were significantly more positive in both 1 Complementary alternative medicine (CAM), also referred to as "complementary therapies", describes those diagnostic and therapeutic techniques outside the realm of what is considered conventional medicine in the United States . BecauseThe CAM Health Belief Questionnaire (CHBQ) is useful in measuring medical students' attitudes toward alternative medicinal theories and practices. Composed of 10 items, the CHBQ requires little time to complete and the survey's complexity is minimized . The CHBst, 2nd, and 3rd year medical students at the University of Birmingham's Medical School, results indicated that females showed a significantly more positive attitude toward CAM than did males [While research regarding attitudes toward alternative medicinal theories is limited, some studies do show that gender does play a role in one's favorability of CAM ,8. Afterid males .th year medical students, 35.4% agreed or strongly agreed that "CAM is a useful supplement to regular medicine" [Studies that examined class level differences vary in outcome and groups surveyed. According to a survey by Baugniet et. al. (2000) given to 61 4edicine" . This saedicine" . Overalledicine" .An earlier study of third year medical students at the University of South Florida (USF) reported that approximately one third the population 28 of 100) surveyed associated the effects of CAM therapy with those of a placebo while 35 participants remained neutral and 37 disagreed [0 surveyest and 2nd year students participated in a study examining CAM and its position in medical education and practice. Chaterji et. al. (2007) reports that 91% of students surveyed agreed that conventional medicine could benefit from CAM teachings and techniques [st and 2nd year students, as on both statements previous, they were in agreement.At the Georgetown University School of Medicine, 266 1chniques . Furtherchniques . The resst and 3rd year medical students surveyed, 27.8% of respondents are knowledgeable in 18 CAM therapies, 40.5% would refer a patient/client for CAM, and 51.4% believe CAM to be an effective means of therapy [st year and 139 3rd year students) were analyzed indicating that 1st year medical students believed CAM to be more effective than 3rd year medical students [st year students demonstrated a greater desire for training in CAM therapies than did 3rd year students [Furnham et. al. (2003) reports that of 311 1 therapy . Differestudents . Similarstudents .st year and 185 2nd year) averaged a mean score of 47.8, equal to a slight positive attitude toward CAM [rd year. Year 1 medical students had a mean score of 46.4, 2nd year students averaged 47.4, and 3rd year students averaged 48.3, representing no significant change between 1st and 3rd year medical students [One study incorporating the CHBQ administered to a total of 355 medical students  provides conflicting results. Thus, it is the purpose of this study to obtain another set of data to compare with previous studies. Based on these findings and the effectiveness of the methods and CHBQ survey, a longitudinal study that can be administered nationally will be designed and its results reported. The specific data provided by this study will also allow variations in attitudes toward CAM between medical institutions and geographical areas to be compared (for example: data from USF vs. data from a northern institution). Furthermore, in examining the changes in attitude toward CAM that occur throughout one's medical education, medical institutions can redesign current or develop new curricula to accommodate such changes.The previous data (not limited to that reviewed in this article) regarding attitudinal differences between MD class levels . The survey was administered to 1st year students at the beginning of their 1st year of medical education (October 2007), 2nd year students during their first month of their respective year in school (October 2007) and 4th year students during their final month of medical school (April 2007). Originally, the study sought out to examine MD students upon their entrance (1st year) and exit (4th year) of medical school, however, access was gained to 2nd year students which accounts for their inclusion in this study. Because the 2nd year students were in the beginning of their 2nd year, and only had 1st year educational backgrounds, they were included in this sample. Third year MD students were beginning their clinical rotations, thus, access to their student cohort was limited and they were excluded. All respondents were students at the USF School of Medicine. Broken down by gender, 40% (38) of the respondents were male; 60% (57) were female.The final collected sample included 95 respondents. Broken down by class, 36 were in their 1st year, 2nd year, and 4th year MD students and 2) between male and female students (P ≤ .05) [see Table In conducting this cross-sectional study, the previously validated and reliability tested Cam Health Belief Questionnaire (CHBQ) was utilized. The 10 question survey which measures medical students' attitudes toward CAM was administered through a web-based survey program that first, second, and fourth year University of South Florida (USF) MD students were granted access to through a standardized group email sent out to the student population. Students were permitted ten days to complete the survey and participation was optional. All participants were required to provide consent prior to starting the survey; confidentiality was maintained. The survey was graded using a 7-point likert scale; 1 meaning absolutely disagree and 7 meaning absolutely agree. Three questions  are negatively worded and scaled in reverse. A maximum score of 70 correlates with a strongly positive attitude toward CAM, a score between 69-36 representative of a positive attitude, and a score of 34-11 correlates negatively with attitudes toward CAM and a 10 is the minimum score possible  was used to examine mean CHBQ scores between 1) 1st, 2nd, and 4th year students were 46.0 ± 7.7, 48.3 ± 7.8, and 37.8 ± 15.7 [see Table st and 4th year students (p = 0.012) and 2nd and 4th year students (p = 0.002). No significant difference was found between 1st and 2nd year students (p = 0.683). When comparing scores based on gender, females (46.1 ± 11.0) scored significantly higher (p = 0.046) than males (41.2 ± 12.2).The mean scores for 1st (46.0 ± 7.7) and 2nd (48.3 ± 7.8) year medical students as compared to 4th year (37.8 ± 15.7) student's attitudes. One study by Lie et. al. (2006), using the CHBQ, found comparable results to those being reported [st year and 185 2nd year medical students shared CHBQ mean scores of 46.4 and 47.4 respectively. The mean CHBQ score for 3rd year medical students, 48.3, indicated no significant change [st and 2nd year medical students are consistent with the scores being reported in this study. Another study by Furnham et. al. (2003) reports that 172 1st year medical students believe CAM to be more effective than 139 3rd year medical students [st and 3rd years of medical school. This study supports these findings [Similar to previous studies involving medical students' attitudes toward CAM ,12, thisst 4.0 ± 7.7 st 4.0 ± 7.7 st and 2nd year students' attitudes are significantly higher, and thus, more positive toward CAM while 4th year students' attitudes represent a slightly positive attitude toward CAM . One primary explanation for this change in attitude can be attributed to the methods in which instruction is given during the first two years of medical school compared to the second two years of medical school. The 1st and 2nd years of the USF curriculum include primarily classroom based lecture, consisting of subjects including but not limited to ethics, humanities, anatomy, physiology, behavioral medicine, pathology, pharmacology, and clinical problem solving. As students enter their fourth year, however, the role of classroom instruction decreases while the amount of time spent in specific medical rotations/clinical increases. It was the hypothesis of this study, that as classroom based learning decreases and student exposure to allopathic techniques and procedures increases , medical students attitudes toward and beliefs in CAM will decrease. The changes in mean CHBQ scores support this hypothesis.Although this study found all three class levels representative of a positive attitude toward CAM, the scores of 1Upon comparing mean scores based on gender, females (46.1 ± 11.0) had significantly more positive attitudes toward CAM than males (41.2 ± 12.2), though both genders represented an overall positive attitude toward CAM. These findings are consistent with results from other studies in which medical students' attitudes toward CAM based on gender were examined ,8. One sOne limitation of this study may be that the attitudes of individual groups entering medical school each year are already more positive toward CAM than the attitudes of those students who entered medical school in previous years. This factor can be discredited as a limitation by readministering the CHBQ to students during each year of their medical training. Furthermore, the health industry in Florida is very different from the current health industries found in both Western and Northern states, a factor which may have also limited the results of this study. Included as part of Florida's health industry, are two specific CAM programs directed toward students. As a minimal limitation, the authors recognize both the AMSA Leadership Retreat for CAM (which has been held in Florida) and the HEART program (at the University of Florida College of Medicine) as minor limitations that may have influenced student's attitudes toward CAM, however, this inference is strictly theoretical. Furthermore, programs similar to these are not limited to Florida  and similar programs are found throughout the country. The authors also recognize the low response rate of medical students in completing the web based CHBQ. However, in a comparative study of web and mail survey response rates, it has been reported that web response rates (with email notification only) are 10% lower than mail survey response rates . This study reports a 21% response rate for email notification only survey instruments. The response rates for our study (email notification only) are higher than the response rates reported in the Kaplowitz, et. al (2004) study. The most important limitation of this study remains that it was conducted at only one medical institution (The University of South Florida) and that its results may not be representative of the attitudes of medical students nationally.st and 2nd year medical students when compared with 4th year medical students gives strong cause for confidence in an increasing interest and positive attitude  toward CAM. This level of interest and the extent to which it changes during medical training, however, remains to be studied, as a longitudinal nationwide survey involving other medical institutions would be more suited for this purpose.Despite these limitations, the significant difference in CHBQ scores reported in this study between both 1The authors declare that they have no competing interests.MS carried out the survey's administration and statistical analysis, CR conceived the study, participated in its design, and drafted the manuscript. All authors read and approved the final manuscript.


The Sheffield Trophoblastic Disease Centre treats about 25 patients with persistent trophoblastic disease each year. A total of 75% of patients are classified as low risk according to the Charing Cross Hospital prognostic scoring system and receive methotrexate (MTX) 50 mg, i.m., on days 1, 3, 5, 7 with folinic acid 7.5 mg orally 24 h after each methotrexate injection. There is a 7-day rest between treatment cycles. Remission is achieved in 85% of cases. Approximately 20% of patients experienced pleuritic chest pain and dyspnoea. We have evaluated prospectively lung function in 16 low-risk patients receiving methotrexate. All patients had pulmonary function tests  performed before and after completed treatment. A mean of 7.5 cycles of MTX were administered (range 4-11). There was a significant reduction in the mean TLCO , but there were no other statistically significant changes. Three patients experienced respiratory symptoms and were found to have a 39%, 28%, and 11% reduction in TLCO from baseline, improving on follow up to pretreatment levels. Low-dose MTX is an effective therapy but may cause troublesome pulmonary toxicity.


These were linked to the Cancer Registry of Norway. A reduced risk of malignant disease was demonstrated among twins for all tumour sites combined; standardized incidence rate (SIR): 0.90 (95% CI 0.85–0.94) in females and 0.95 (95% Cl 0.90–0.99) in males. In both sexes, we observed a significant reduced incidence of malignant melanomas of the skin. The incidence of colorectal cancer tended to be reduced for both sexes. In females, the incidence of tumours of the central nervous system and lungs were reduced. We consider our findings are real, but cannot explain them. © 2001 Cancer Research Campaign


The oral health condition of individuals with special health care needs have been reported in literature to be influenced by various sociodemographic factors, including living conditions and severity of impairment. This study was carried out to determine the oral health status and treatment needs of children and young adults attending a day institution for those with special needs.This study was carried out as part of an oral health screening program organized by the institution and consent was obtained from parents and guardians before the screening. All information was supplied by the parents during the screening using a questionnaire completed by the dentist. Oral examination was carried out on all consenting subjects in attendance on the days of screening in the school clinic with parents and teachers in attendance, using standard World Health Organisation oral health indices to assess dental caries, oral hygiene status, malocclusion and other oral health parameters.Fifty-four subjects aged 3–26 years (mean 12.28 ± 6.82 years) and comprising 72.2% males and 27.8% females participated in the study. Over 90% were from parents of high and middle level educational background. Thirty-six (66.7%) were caries free, with a mean dmft score of 0.7 ± 1.77 and mean DMFT score of 0.4 ± 1.44 with no significant difference across gender (p = 0.5) and parents' educational status (p = 0.43). The mean OHI-S of the total population in this study was 1.36 ± 0.16. Females had a mean score of 0.88 ± 1.10 while males had a mean score of 1.55 ± 1.24 with no significant difference (p = 0.6). Twenty-five (46.3%) had good oral hygiene, 17 (31.5%) had fair oral hygiene and 12 (22.2%) had poor oral hygiene, with no significant difference across gender (p = 1.11) and age groups (p = 0.07). Fifteen (27.8%) had gingivitis with no significant difference across age groups (p = 0.17). Forty-five (83.3%) had Angle's class I malocclusion, 6(11.1%) class II and 3 (5.6%) class III. Chronologic enamel hypoplasia was found in 9 (16.7%) of the total population. Up to 53.7% of the total population will require oral prophylaxis, 33.3% required restorations on their posterior teeth and 12.9% required veneers for labial facing of hypoplastic enamel.The subjects in this study had a high prevalence of dental caries and need for restorative care. They would benefit from parental education on diet modification, improvement of oral hygiene practices and regular dental visits. Individuals with special health care needs have been reported in literature to have poorer oral hygiene and periodontal status, more untreated caries and fewer remaining teeth -3. They Their oral health condition may be influenced by age, severity of impairment and living conditions. Individuals with special needs may have great limitations in oral hygiene performance due to their potential motor, sensory and intellectual disabilities -7, and sPoor oral health conditions have also been linked to low socio-economic status. Poor and nearly poor children with special health care needs and those with greater limitations attributable to disability were more likely to have unmet dental care needs . EarlierThe study population consisted of children and young adults attending a private day centre for individuals with special needs in an urban area of Lagos. A similar study was carried out earlier, using the same criteria and tools, on children and young adults attending three public schools for individuals with special needs in the same area in Lagos . At the One of the authors (FAO), carried out the oral examination on all the subjects in the school clinic using natural light. They were examined for the following parameters using the World Health Organization Oral Health Survey Basic Methods :using the decayed, missing and filled teeth (dmft) index for primary (0–5 years) and early mixed dentition (6–19 years) and Decayed, Missing and Filled Teeth (DMFT) index for late mixed (11–15 years) and permanent dentitions (16 years and above). A tooth was considered decayed when there was frank carious cavitation on any surface of the tooth. A tooth was classified as missing in the index if it was extracted due to caries. A tooth was classified as filled if it had a restoration for a carious lesion. Exfoliated teeth in the primary and mixed dentition, unerupted and those extracted for other reasons apart from caries were not included in the indices.using the Simplified Oral Hygiene Index (OHI-S) of Greene and Vermillon. The oral hygiene of each child was classified as 'good' when the OHI-S score was 0–0.9, 'fair' when it was 1.0–1.9 and 'poor' when it was 2.0 up to 6.Angle's classification of occlusion was used to classify malocclusion. Crowding, spacing and anterior open bite were also recorded.Consistent discoloured malformations on teeth of the same series in at least two quadrants.A tooth was classified as missing if it had not erupted after six months of its expected eruption date.A tooth was classified as retained if it was still in the arch after six months of its expected date of exfoliation.Fracture of anterior teeth was also recorded.Findings were communicated to the parents/guardians, appropriate oral health education given and written referrals given to the dental clinic where necessary.Data obtained were analyzed with the health statistical software Epi info version 6 . Chi squSixty-six (69.5%) subjects responded to the call for screening. Twelve were either ill or very uncooperative so were excluded from the study, leaving 54 (56.8%) subjects, aged 3–26 years (mean age 12.28 ± 6.82 years) who were examined. There were 37 (72.2%) males and 15 (27.8%) females. Thirty five (64.8%) were from parents of high educational level, 18 (33.3%) were from the middle and only 1 (1.9%) was from the low educational level and so grouped with the middle educational level. There were 13 (24%) subjects in each of the 0–5 and 6–10 years age groups, 10 (18.6%) in the 11–15 years age group and 9 (16.7%) each in the 16–20 and 21 ≤ years age groups (Table Thirty-six (66.7%) were caries free. The mean dmft of the primary and early mixed dentition years was 0.7 ± 1.77 while the mean DMFT of the late mixed dentition and permanent dentition was 0.4 ± 1.44 as shown in Table The mean OHI-S of the total population in this study was 1.36 ± 0.16. Females had a mean score of 0.88 ± 1.10 while males had 1.55 ± 1.24 with no significant sex difference .Twenty-five 46.3%) had good oral hygiene, 17 (31.5%) had fair oral hygiene and 12 (22.2%) had poor oral hygiene, with no significant difference across sex (p = 1.11), age groups (p = 0.07) and parents' educational level (p = 0.17)  had gingivitis with no significant difference across gender , parents' educational level  and age groups . Gingivitis was significantly present in subjects with seizure disorders, learning disability, Down syndrome and autism . Forty-five (83.3%) had Angle's class I malocclusion, 6(11.1%) class II and 3 (5.6%) class III. Five (24%) of those with cerebral palsy had class II malocclusion while 3 (60%) of those with Down syndrome had class III malocclusion. Chronologic enamel hypoplasia was found in 9 (16.7%) of the total population, with no significant difference across the different disabilities  and age groups . Anterior open bite was seen in 9.3%, crowding in 12.9%, fractured anterior teeth (3.7%) and retained teeth (3.7%) with no significant difference across age group, gender and parents' education level. Up to 53.7% of the total population will require oral prophylaxis, 33.3% required restorations on their posterior teeth and 12.9% required veneers for labial facing of hypoplastic enamel  seen in an earlier study from the same environment and of comparable age range . It is eThe majority of the subjects were caries-free, although the proportion of caries-free subjects was relatively low compared with that of subjects with special needs in public schools (93%). Some authors have however reported a lower caries prevalence in children with disabilities compared with those without disabilities ,20. The Only two children have had an amalgam and Glass Ionomer Cement restorations. Although this proportion who had received dental care is indeed small, it is encouraging because previous surveys in this environment showed no index of restorative care in the large population studied .In contrast to dental caries, almost half of the subjects in this study (46.3%) had good oral hygiene compared with lower proportions of those in earlier studies in this environment and elsewhere but among children from parents of lower educational background ,15,24. TA high prevalence of unmet needs is still evident in this study despite the educational background of the parents and the fact that the school and residence of the subjects are located in an urban area of the state. Other studies in developed countries have shown that dental care is the most prevalent unmet health care need for children with special health care needs -3,27. ThMore than half of the subjects with Down syndrome (DS) had class III malocclusion which is similar to reports from previous studies on those with DS . Class IPrimary health care providers may influence access to dental care by oral health assessment and prompt dental referral ,37. One There was a high prevalence of dental caries and need for restorative care among the subjects in this study. Their oral hygiene was better than that of subjects attending public schools from previous studies. It is recommended that regular contact be made with parents and caregivers and educated on the need for diet modification, improvement in oral hygiene and regular dental visits for their wards.The authors declare that they have no competing interests.FAO conceived of the idea, drafted the questionnaire, carried out the oral examination, analyzed the data and took part in the draft of the manuscript. YO participated in the draft of the manuscript.The pre-publication history for this paper can be accessed here:


To study the treatment of ureteric stones by HO:YAG laser lithotripsy and pneumatic lithotripsy and to evaluate the results of the two treatment modalities to assess effectiveness and complications.Over 1-year period, a total of 79 patients with 82 ureteral stones were randomized into two groups. In group 1  ureteroscopic HO:YAG laser lithotripsy was performed using a rigid 8 Fr-ureteroscope (LL group). In group 2  pneumatic lithotripsy was performed in like manner. Efficacy safety and complications in both groups were analyzed.A total of 79 patients with 82 calculi were treated. Two cases in LL group and one in PL group had bilateral ureteral stones. Mean stone size was 12.07 mm in LL group and 10.2 mm in PL group. Stones located in lower ureter in 30 cases on LL group and 29 cases in PL group. Proximal migration of stone occurred in 1 case on LL group and in 3 cases on PL group. Successful fragmentation occurred in 37 cases on LL group and in 30 cases on PL group. Stone-free rate after 1 month in the base of Kidney Ureter Bladder (KUB) and sonography was 95% in LL group and 80.5% in PL group. Ureteral perforation, urinoma, and urosepsis were not seen in both groups.HO:YAG laser has advantages over PL in high efficacy of stone fragmentation and a low-retrograde migration of ureteral stone treatment. Other complication of ureteral stone treatment with LL and PL are the same and very rare. Endoscopic management of ureteral calculi is one of the most important therapies in academic centers. DifferenThere are a few studies to comparison of these two techniques and the aim of this study is evaluation of the results of Trans Ureter Litotripsy (TUL) with these two methods according to their fragmentation, complication, proximal migration, and stone-free rates.In a clinical trial study science February 2003 to February 2004, a total of 79 patients with 82 ureteral stones were randomized in two groups. In group 1 , Ho:YAG laser lithotripsy was performed by a rigid 8.5-Fr Wolf ureteroscope (LL group) and in group 2 , the procedure was performed with a pneumatic lithoclast (PL group) at the same size of ureteroscope.2 and t-test.Patients were included in the study when they had ureteral stone which not passed in 2 weeks, presence of hydronephrosis, and failed Extracorporeal Shock Wave Litotripsy (ESWL). Stones more than 1.5 cm were excluded. In all patients, urinalysis, urine culture, KUB, ultrasonography, and intravenous pyelography were performed before TUL. Preoperatively, all patients were received 1 g ampicillin and 80 mg gentamycin intravenously. The patients were placed in lithotomy position and cystoscopy was performed before the TUL. A guide wire was inserted into the ureteral orifice and ureteroscope (8.5-9 Fr wolf) was used in all patients. The stones were fragmented by a 0.8-mm Swiss lithoclast probe with the ballistic energy at a rate of 12 Hz or Ho:YAG laser (DECA 20 watt's Smart 2001) with end-firing probe 365 μm, 0.5-1 joule energy in frequency of 5-10 Hz. The indications for stenting were ureteral edema secondary to an impacted calculus, iatrogenic trauma, and residual stone burden. KUB and ultrasonography were done 1, 3, and 30 days after the procedure. Results were analyzed with Pearson χA total of 79 patients with 82 ureteral calculi were treated. Forty cases treated with pneumatic lithotripsy (PL group) and 39 cases with HO:YAG laser lithotripsy (LL group).From 40 patients on PL group, 29 cases were males with mean age of 38.5 ± 6.4 years; and 11 cases were females with mean age of 42.5 ± 4.3 years, from 39 patients on LL group, 28 cases were males with mean age of 35.71 ± 41 years and 11 cases were females with mean age of 51.09 ± 5.1 years. Two cases in LL group and one in PL group had bilateral ureteral stones.Mean stone size was 12.07 ± 2.1 mm in LL group and 10.2 ± 2.8 mm in PL group with a range of 6-15 mm in both groups. Stones located in lower ureter (below the iliac crest on the base of KUB) in 30 cases of LL group and 29 cases of PL group. In other patients stones location was in the upper ureter (above the iliac crest).P = 0.6). Successful stone fragmentation occurred in 37 cases (90.2%) on LL group and in 30 cases (73.20%) on PL group. In Pearson χ2-test, this difference was significant (P = 0.46).Proximal stone migration occurred in 1 case of LL group (2.4%) and in 3 cases of PL group (7.3%), that was insignificant (P = 0.043) and this was significant also.Stone-free rate after 1 month in the base of KUB and ultrasonography was 95% in LL group and 80.5% in PL group . In Yinghao study, this was 95.7 and 69.7% for LL and PL groups, respectively.[et al, reported stone fragmentation with PL was better than LL. Their laser was alexandrite laser.[Under direct vision stone fragmentation with LL was better than PL, especially when stone surface was smooth and hard. In our study, stone fragmentation was 90.2% in LL group and 73.2% in PL group (te laser. ThereforP = 0.6).In this study, eight cases in PL and two in LL groups had fragments larger than 5 mm. After 1 month follow-up, three cases in PL and both of two cases in LL group, become stone free. However, retreatment performed in the rest of patients. Proximal stone migration is the most disadvantage of the pneumatic lithotripsy and reported in the 2-17% of cases.[Proximal migration is related with dilatation of proximal ureter, size and hardness of stone, severity of stone impaction, and pressure of irrigation fluid.Overall stone-free rate after 1 month without any retreatment procedure were 95.1% in LL and 80.5% in PL groups. This result is comparable with other studies.1565In our setting major complications such as ureteral perforation urinoma and urosepsis were not seen on both groups.It seems that Ho:YAG laser has advantages over pneumatic lithotripsy in high efficacy of stone fragmentation and a slightly low-retrograde migration in the treatment of ureteral stones.Overall, stone-free rate in Ho:YAG laser lithotripsy is better than pneumatic lithotripsy. Other complications of ureteral stone treatment with Ho:YAG and pneumatic lithotripsy are the same and very rare. Further study is recommended in this field.


An increase in the number of overweight and obese subjects in the general population has been observed.The aim of this study was to determine the prevalence of overweight and obese subjects in the general population and its association with undiagnosed pathologies, such as diabetes mellitus [DM] and hypertension [HT], by taking age, gender and place of residence [rural or urban] into account.A cross-sectional population-based survey was conducted in Castellón, East Spain in 2005–2006. The sample included 2,062 participants aged 18–94 years. Weight, height, blood pressure and glycaemia values were recorded, and information about gender, age and place of residence was obtained. Overweight, obesity, and undiagnosed HT and DM prevalences were calculated. Multiple regression analyses were done to assess the association of overweight/obesity with undiagnosed HT and DM by adjusting for age, gender and place of residence.The overall overweight, obesity, and undiagnosed HT and DM prevalences were 39.9% [95% CI:37.3–42.0], 25.9% [95% CI:24.0–27.9], 9.0% [95% CI:7.8–10.4] and 12.6% [95% CI:11.2–14.1], respectively. We identified various independent risk factors; those relating to overweight were increasing age, male gender and rural residential area, while that relating to obesity was increasing age. Compared to normal weight adults, the Relative Prevalence Ratio (RPR) for subjects who were overweight and had HT was 2.00 [95% CI:1.21–3.32]; that for obesity and HT was 1.91 [95% CI:1.48–2.46], and it was 1.50 [95% CI:1.25–1.81] for obesity and DM.Overweight and obesity prevalences, and their association with undiagnosed DM and HT, are high in our study population. In the last two decades, the prevalence of overweight and obesity has increased worldwide , mainly In Europe, obesity prevalence rates have increased by 30% since the mid 80's , and areIt is suspected that Spain has the highest prevalences in the European Union for both genders , and a gThis increased prevalence of obesity has also been associated with the increased risk of numerous comorbidities like hypertension [HT] -10 and dThis study aims to determine the overweight and obesity prevalences in the general adult population and their association with undiagnosed pathologies [DM and HT] by taking age, gender and area of residence [rural or urban] into account.A cross-sectional study was done in a general population of Castellón, Eastern Spain in 2005–2006.The study population included subjects aged 18 years or older and who were undiagnosed with DM and HT.In the study area [Castellón], there are 3 Health Districts with a population of 431,093 subjects aged 18 years or older .Of this population we selected 8,400 subjects by a random, stratified, multi-stage sampling technique in which the first-stage units were Primary Health Care Centres. Those with diagnosed HT or/and DM were excluded .The other 4,898 subjects [undiagnosed of HT and DM] were located by telephone and invited to participate in the study. This sample size allows us an accuracy of 99.0% for an expected obesity prevalence of 15.5% . The subThe final sample included 2,062 adults , 643 men [31.18%] and 1419 women [68.82%]. This sample size allows us to an accuracy of 98.5% for an expected obesity prevalence of 15.5% [Participants' ages ranged from 18 to 94 years. The mean age of men and women was 58.00 ± 14.73 and 54.83 ± 14.52 respectively. Written consent to participate in the study was obtained from all the participants.2]. Those individuals whose BMI was between 25.0 and 29.9 kg/m2 were considered overweight, while those with a BMI equal or over 30 kg/m2 were classified as obese [Specially trained professionals [clinical specialist pharmacists] conducted an anthropometric study with the participants. This study recorded weight [kg], height, and calculated body mass index [BMI] [kg/mas obese .The participants' blood pressure was consecutively taken three times and the arithmetic mean was calculated. Diagnosing HT was conducted by following the World Health Organization's [WHO] criteria  .Capillary glycaemia [CG] was also determined in all the participants. Only in those participants whose CG value exceeded 106 mg/dl [if he/she had not eaten breakfast] or 140 mg/dl [if he/she had eaten breakfast], was venous glycaemia [VG] also dertmined by taking blood samples [before they ate breakfast in all the cases]. Diagnosing DM was conducted based on VG by following the American Diabetes Association criteria [DM: subjects with a VG value before eating breakfast of > 126 g/dl] .Information about the participants' age, gender, and place of residence was obtained through interviews. We distinguished between places of residence as follows: rural areas with < 10,000 inhabitants; urban: areas with ≥ 10,000 inhabitants.Overweight, obesity and undiagnosed HT and DM [who were not on HT/DM treatment and whose blood pressure/VG exceeded the limits stated] prevalences were calculated with their corresponding confidence intervals [CI 95%]. Comparisons were done with the Chi-square test [p ≤ 0.05].Given the sample's lack of representativeness, prevalences were estimated in the adult population of the province of Castellón by the direct standardisation procedure for samples in terms of age and gender  by takinIn order to determine the relationship of being overweight/obese with age, gender, place of residence and having undiagnosed DM/HT, a multiple regression logistic analysis was done by calculating the relative prevalence ratios [RPR], which was adjusted for age, gender and place of residence, as well as their corresponding CI 95%. Statistical analyses were done with the SPSS v.15 software.The total overweight and obesity prevalences obtained were 39.9% [95% CI:37.7–42.0] and 25.9% [95% CI:24.0–27.9], respectively. By extrapolating to the adult population of Castellón, which had been adjusted for age and gender, the estimated overweight and obesity prevalences were 38.0% [95% CI:37.8–38.2] and 16.7% [95% CI:16.6–16.9] . By extrapolating to the adult population of Castellón, the estimated HT prevalence was 5.5% [95% CI:5.4–5.6] Table .Of all the participants, 260 had DM with a prevalence of 12.6% [95% CI:11.2–14.1]. By extrapolating to the adult population of Castellón, the estimated DM prevalence was 7.3% [95% CI:7.2–7.4] .Table The overall overweight and obesity prevalences for men were 50.4% [95% CI:46.5–54.2] and 23.8% [95% CI:20.6–27.3], and 34.9% [95% CI:32.5–37.5] and 26.8% [95% CI:24.6–29.2] for women. By extrapolating to the adult population of Castellón, the estimated overweight and obesity prevalences for men were 46.8% [95% CI:46.6–47.1] and 16.0% [95% CI:15.8–16.2], and 29.3% [95% CI:29.1–29.6] and 17.4% [95% CI:17.3–17.6] for women. Men of all ages tended to have higher prevalence of overweight than women. The prevalence of obesity tended to be higher in women after age 45 years and the difference was significantly higher in the oldest age group  with an RPR of 1.57 [95% CI:1.19–2.07] compared with those living in an urban area. For obesity, the prevalence was somewhat higher for subjects living in a rural area than for those living in an urban area, although no statistically significant differences were found  [Table Table There was a tendency in all age groups for more undiagnosed HT and DM in men compared to women.As shown in Table We observed a high prevalence of overweight and obesity in a Mediterranean region, and this result coincides with other studies . The obeBoth overweight and obesity appears to increase with age. This may be due to people of this age living a more sedentary lifestyle. Besides, a series of physiological alterations take place with age which increase body weight .The overweight prevalence generally tended to be higher in men than in women. However, the obesity prevalence tended to be higher in women than in men. This indicates that obesity prevention measures must address men and women equally.Like other Spanish studies , higher These high overweight and obesity prevalences could be responsible for a possible epidemic of associated adverse effects ,3. SubjeWe also noted that the undiagnosed DM prevalence was statistically and significantly higher in obese subjects than in normal-weight subjects. BMI is an indicator of obesity and would be a possible indicator of the risk of DM ,26.We also observed that the prevalence of this pathology increased significantly with age, as other authors discovered . TherefoThis study is not without its limitations. The main limitation is the sample's lack of representativeness produced by the fact that the response rate was much more elevated in women than in men. Also, for those cases in which hyperglycaemia was absent, a diagnosis of DM was not confirmed with a second test. The Oral Glucose Tolerance Test was not done; therefore the prevalence of undiagnosed DM could be underestimated.This study provides strong evidence for overweight and obesity prevalences in the general population. Besides, it demonstrates the important role that obesity plays as a main modifiable risk factor in the development of comorbidities like HT and DM and its use in early diagnoses.BMI: Body Mass Index; CG: Capillary glycaemia; CI: Confidence Intervals; CIBER: Networked Biomedical Research Centre; DM: Diabetes Mellitus; HT: Hypertension; kg: Kilograms; RPR: Relative Prevalence Ratios; WHO: World Health Organization.The authors declare that they have no competing interests.AHM had the original idea for the work and organized the study. ESI contributed to the study design. FBM provided expert advice on study organization and contributed to the writing of the manuscript. MTMH and JVGM contributed by interpreting the results and helped to write the manuscript. NGC and MMSV did the data analysis and contributed to the writing of the paper. All the authors read and approved the final manuscript.


Knowledge of the prevalence of type-specific human papillomavirus (HPV) infections is necessary to predict the expected, and to monitor the actual, impact of HPV immunisation and to design effective screening strategies for vaccinated populations.N=4719), CIN3/CGIN and cervical cancer biopsies (N=1515) were obtained from sites throughout England, anonymised and tested for HPV DNA using the Linear Array typing system (Roche).Residual specimens of cervical cytology  type) was 76% in squamous cell carcinomas, 82% in adeno/adenosquamous carcinomas and 63% and 91% in CIN3 and CGIN, respectively. Of all HR HPV-infected women undergoing cytology, non-vaccine HPV types only were found in over 60% of those with mild dyskaryosis or below, and in <20% of those with cancer. In women of all ages undergoing screening, HR HPV prevalence was 16% and HPV 16 and/or 18 prevalence was 5%.Pre-immunisation, high-grade cervical disease in England was predominantly associated with HPV 16 and/or 18, which promises a high impact from HPV immunisation in due course. Second-generation vaccines and screening strategies need to consider the best ways to detect and prevent disease due to the remaining HR HPV types. A wide range of international studies have found approximately 70% of cervical cancers to be associated with human papillomavirus (HPV) types 16 or 18, and therefore preventable by currently available HPV vaccines  and for each disease phase , prior to any impact of the National HPV Immunisation Programme.Two sample types were included in the study: (i) tissue sections from routinely obtained diagnostic biopsies of cervical cancers and high-grade precancerous lesions archived in NHS pathology laboratories, and (ii) residual liquid-based cytology (LBC) samples from women attending for cervical screening. Samples were obtained from eight participating NHS pathology laboratories in England. Three laboratories ) provided both tissue sections and LBC samples, three laboratories  provided tissue sections only and two laboratories  provided LBC samples only.Cancer and CIN3/CGIN biopsies Sections from archived blocks of cervical intraepithelial neoplasia 3 (CIN3), cervical glandular intraepithelial neoplasia (CGIN), squamous cell carcinoma (SCC) and adenocarcinoma (including adeno-squamous carcinoma) (ADC) tissue were collected from the six participating sites between June 2006 and July 2008. Cases were deemed suitable for inclusion in the study after confirmation of the pathological diagnosis by an experienced subspecialist gynaecological histopathologist at each of the six participating centres. The biopsies of cancer had been originally embedded between 1986 and 2008; 96% had been embedded since 2000. The CIN3 and CGIN biopsies had been collected between 2000 and 2008. Approximately six 10 μ thick sections from each tissue block were placed in a 1.6 ml Eppendorf tube. A further section was stained and re-examined by the pathologist to confirm the presence of diseased tissue. To prevent cross-contamination from residual wax from the previous block, the microtome blade was cleaned with ethanol between blocks. Tissue sections were sent to the Department of Clinical Virology, Central Manchester University Hospitals NHS Foundation Trust for HPV genotyping. The following data were collected on a study record for each sample and sent to Manchester: month and year of birth, date of sample collection, type of lesion  and histology result.μl of methanol. Tubes were vortexed and incubated at 56 °C for 30 min followed by centrifugation at 13 000 r.p.m. for 1 min after which the octane layer was removed using a fine-tipped Pasteur pipette. The tissue pellet was then washed with 1.0 ml of ethanol, centrifuged as before and the ethanol removed. The tubes were then left at 56 °C for 30–45 min to evaporate off residual ethanol. The tissue was digested in 600–1000 μl (depending on size of pellet) of proteinase K lysis buffer containing 10 mM Tris/HCl buffer pH 8.3; 1 mM EDTA, 0.5% Triton X-100, 0.002% SDS and 250 μg ml–1 of proteinase K. Tissue digestion was carried out at 56 °C for 72 h with constant agitation. DNA was then extracted from a 200 μl aliquot of digested tissue using the Roche MagNA Pure  automated extraction system with an elution volume of 100 μl.Tissue sections were de-waxed by adding a 1 ml of octane to each Eppendorf tube followed by 75 LBC samples Residual LBC samples were collected prospectively from women undergoing cervical screening between October 2007 and January 2009 at five participating sites. Stratified sampling was used to obtain sufficient high-risk (HR) HPV-positive samples to analyse type distribution within each combination of age-band and cytology grade, thus a disproportionately higher number of older women and those with more severe cytology outcomes were sampled. Before inclusion in the study, samples were collected and handled according to local protocols: Thinprep and Surepath LBC systems were both in use, with samples stored at ambient temperatures while awaiting cytological examination. After completion of cytology, residual LBC samples that matched an unmet age-group and grade (according to the site’s stratified sampling frame) were anonymised and sent to the Health Protection Agency (HPA) Virus Reference Department for HPV genotyping. For Surepath samples, the tubes (enriched sample), not the vials, were used as the vials were found to have an inadequate amount of cellular material. The tube approximated to levels found using Thinprep, although with more variability. The following data were collected on a study record for each sample and sent to the HPA: month and year of birth, cytology result, date of sample, outward postcode , biopsy taken (and histology result if applicable).μl of cold sterile phosphate-buffered saline. After a lysis step (addition of 40 μl Qiagen Protease and 265 μl Qiagen Buffer AL containing guanidine hydrochloride) samples were stored at −25 °C before nucleic acid extraction. Nucleic acid extraction was conducted on the BioRobot Universal platform  using the QIAamp DNA Blood BioRobot MDx kit and the extraction protocol QIAamp ‘One for All UNIV rcV23’.1 ml of each LBC sample was centrifuged for 5 min at 13 000 r.p.m. and the cellular pellet suspended in 300 μl aliquot of the extracted DNA from the biopsy samples and a 10 μl aliquot of the extracted DNA from the LBC samples  test ) was used for the PCR reaction stage of the Roche LA test. The Roche LA test was used according to manufacturer’s instructions, with the exception of the sample extraction methods described above. Individual oligonucleotide capture probes enabled identification of 37 HR and low-risk HPV genotypes and the β-globin amplicon acted as a control for cell adequacy, extraction and amplification. Identification of HPV 52 in this system was possible by the use of a cross-reactive oligonucleotide probe (52 M) that hybridises with HPV genotypes 33, 35, 52 and 58. The presence of HPV 52 was therefore inferred by reactivity to the 52 M probe with no reactivity to the individual oligonucleotide probes representing HPV 33, 35 and 58. The presence of a co-infection of 52 with 33, 35 or 58 would not have identified HPV 52. The HPV-positive and -negative controls provided within the Roche LA test kit were included for every 46 tissue section samples and every 22 LBC samples tested. The time from LBC sample collection to receipt at HPA  was not negatively associated with detection of β-globin or HPV DNA: this variable was not considered further.A 50 β-globin low control band were recorded.Following an assessment of type-specific assay reproducibility (details in β-globin negative or in which no HR HPV DNA was detected using Roche LA test (β-globin control), however, it amplifies a much smaller fragment of the DNA than the Roche LA assay making it less likely to be affected by DNA fragmentation, which may occur during tissue fixation. A 5 μl aliquot of extracted DNA was amplified and detected according to manufacturer’s instructions.For tissue section samples only, additional analysis using the Inno-Lipa Extra  genotyping assay was performed on those samples that were  LA test . The Innβ-globin result by either Roche LA or Inno-Lipa Extra. Only samples with a valid HPV typing result , while the LBC samples were from 2452 women without cervical disease, 1051 with borderline, 697 with mild, 276 with moderate and 243 with severe dyskaryosis. The mean (s.d.) age of the women at biopsy was 31.9 (8.1), 49.7 (16.1), 35.4 (7.8) and 45.5 (14.1) years for CIN3, SCC, CGIN and ADC, respectively. The numbers of samples included in the study, by participating site, are available in The prevalence of each of our defined categories of HR HPV, by age-group and disease-grade, from normal to cancer, are shown in In biopsies from high-grade cervical abnormalities, HR HPV were detected in 94.2% (95% CI 87.7–97.3%) and 98.1% (95% CI 86.4–99.8%) of CIN3 and CGIN, respectively. HPV 16 and/or 18 were detected in 63.2% (95% CI 57.0–69.1%) and 90.7% (95% CI 85.0–94.4%) of CIN3 and CGIN, respectively, and were the only HR types detected in 53.9% (95% CI 48.7–59.0%) and 83.3% (95% CI 61.1–94.1%), of CIN3 and CGIN, respectively .n=226) and 62.6% (n=149), respectively. Women of all ages with moderate dyskaryosis also had high prevalence of HR HPV and of HPV 16 and/or 18 infection   and 5.1% (95% CI 3.3–8.0%), respectively . The prectively) . With inctively) .χ2P>0.05). Further exploration of HR HPV prevalence in young women (25–29 years) with normal cytology results also found no significant differences between regions (data not shown).Prevalence of HR HPV by region ranged from 13.3% in Gloucestershire to 20.1% in London, while prevalence of HPV 16 and/or 18 ranged from 4.2% in Gateshead to 6.5% in London . DiffereHPV 16 was the most commonly detected type in all cervical grades apart from CGIN wherein HPV 18 was the most commonly detected type. The four next most frequently identified HPV types in the SCC biopsy samples (in single or multiple infections) were, in descending order, HPV 33, 45, 52 and 31. The prevalence of these six types in cancer, precancerous lesions and normal cytology samples is shown in The four HR types found most commonly in SCC biopsy samples, after HPV 16 and 18, showed a general pattern of increasing prevalence with increasing grade of disease although all, except HPV 45, had higher prevalence in severe grade lesions and CIN3 than in cervical cancers . The ageThe prevalence, in the absence of HPV 16 or 18, in SCC and ADC of selected HPV types from the A9 and A7 species, closely related to HPV 16 and 18 respectively, is shown in n=46, 0.8% weighted prevalence). Of these samples, 34.8% (n=16) had normal cytology, 30.4% (n=14) had borderline and 34.8% (n=16) had mild dyskaryosis. No women with moderate or severe dyskaryosis were found to have HPV 6 and/or 11 without a HR HPV type. Overall (irrespective of other types), the age-weighted (to NHS CSP population) prevalence of HPV 6 and/or 11 was 0.9% (95% CI 0.6–1.5%) in normal samples, 0.1% (95% CI 0.08–0.14%) in borderline samples, 0.1% (95% CI 0.05–0.3%) in mild dyskaryotic samples, 0.02%  in moderate dyskaryotic and 0.01%  in severe dyskaryotic samples. In women with normal cytology, the weighted prevalence of HPV 6 was 0.8% and HPV 11 was 0.2%  samples and biopsies of cervical cancers and high-grade lesions, collected from eight sites, to describe type-specific HPV prevalence in women in England, across the full spectrum of cervical pathology, from normal to cancer.Assuming the relative frequency of SCC and ADC in our study are representative of all cervical cancers, our results suggest that at least 73% of cervical cancers in England are potentially preventable by type-specific protection from HPV 16/18 immunisation , rising to around 77% if co-infection with other HR types is never the causative HPV infection. Similarly, at least 56% and up to 65% of CIN3/CGIN are potentially preventable by HPV 16/18 immunisation. The difference between the percentage of cases with HPV 16 and/or 18 only (minimum attributable fraction), and the percentage with HPV 16 and/or 18 and another HR HPV (probable attributable fraction), indicates the potential for unmasking of disease caused by other HR HPV types if HPV 16 and 18 infections are prevented. By the same reckoning, between 32 and 49% of the moderate and 40 and 64% of the severe dyskaryotic cervical abnormalities identified by cervical screening may be preventable by HPV 16/18 immunisation.HPV 31 and 45 were each associated  with an additional 3% of SCC and 2% of ADC. As there is some evidence of cross-protection from current HPV 16/18 vaccines against infection and disease due to these types and HPV 33, 52 and 58, which are closely related to HPV 16 or HPV 18 (N=222) and 91% of ADC (N=47) diagnosed between 2000 and 2006. Three other studies have looked at fewer than 50 cases each . Our prevalence estimates do not provide data on the 20% of the population who do not regularly attend for screening , and whoThis is the first study to describe comprehensively type-specific HPV prevalence across England, including samples from women with normal cytology and all stages of cervical disease in order to be able to estimate the proportion of disease potentially preventable, nationally, by immunisation. These data will be used in models to assess the likely benefit from HPV immunisation (


Acetonitrile was used to simultaneously deproteinize rat plasma and extract metformin. The assay was linear in the concentration range of 0.33 μg-16.6 μg/ml with co-efficient of correlation 0.994. The retention time was 4.7 min. The method was found to be precise (% CV < 15%), accurate and suitable for pharmacokinetic study of orally administered metformin in rats.A simple reverse phase high-performance liquid chromatographic method has been developed for determining the concentration of metformin in rat plasma. The method employs C As itsMetformin was a generous gift by Zim Laboratories Ltd., Nagpur, India. Acetonitrile (HPLC grade) and ammonium acetate (AR grade), were purchased from Ranbaxy Chemicals, India (Rankem). Freshly double distilled, deionised water filtered through 0.45 μ nylon  filter in Millipore unit (USA) was used throughout the experiments. Standard solution of metformin (1 mg/ml) was prepared in mobile phase and kept at 4-8°. Fresh stock solutions were prepared every week. The serial dilutions of stock solution were made to obtain working standard solutions of 0.5, 1.0, 2.0, 5.0, 10.0, 15.0, 20.0, 50.0 μg/ml strength. These solutions were freshly prepared every week and stored at 4-8°.ad libitum. The Institutional Animal Ethics Committee (IAEC) approved the use of animals. Whenever required, blood (1.0 ml) was withdrawn by micro-capillary technique from retro-orbital plexusSprague-Dawley rats (220-250 g) of either sex housed under the controlled conditions of temperature and humidity  with dark/light (12/12 h) cycle and received a standard pelleted diet  and water 18 column  at a rate of 1 ml/min. Detection of metformin was carried out at 236 nm at ambient temperature. The data were interpreted using CSW (chromatographic work station) data acquisition software. The chromatographic study was carried out using Shimadzu 10AT/Vp HPLC system, having in-built degasser unit (DGU 14A), mixer unit (FCV 10AL) attached to solvent delivery module with low-pressure gradient pump (LC 10AT), Rheodyne injector port  and UV/Vis detector (SPD 10A). The mobile phase was prepared by mixing 0.15 M ammonium acetate (pH 5.5) and acetonitrile in the ratio of 90:10; it was filtered through 0.45 μ membrane filter. The elution was carried out on C Calibration curve was established by spiking rat plasma (500 μl) with known amount of metformin to obtain 0.33, 1.66, 3.33, 5.0, 6.66, 16.60 μg/ml. The lowest concentration of the analyte that gives at least 5 times the response as compared with blank was considered as the limit of quantitation (LOQ). Quantitation of metformin in rat plasma was done by reading the analyte response against the calibration curve.In order to determine the intra-day and inter-day accuracy and precision, the concentration of metformin present in five replicates of plasma spiked with 3.33, 5.00, 6.66 μg/ml metformin was estimated by HPLC within a day or on three consecutive days. 85-115% accuracy and coefficient of variation values < 15% except at LLOQ (accuracy 80-120% and CV ≤20%) were considered acceptable.Recovery of metformin from plasma was estimated at 3.33, 5.00, 6.66 μg/ml concentrations by comparing peak area of spiked plasma standards with those of corresponding plain standards containing the corresponding concentration in mobile phase that represent 100% recovery.Analyte stability in plasma was tested at 3.33, 5.00, 6.66 μg/ml concentrations in replicates of three for one freeze-thaw cycle, long term (30 days), and short term (24 h) stabilities. Initially, a plasma standard containing above concentrations were prepared and divided into four fractions. One fraction was analyzed immediately and its area was noted. The other three fractions were stored, two at -20°, and remaining at room temperature. The sample kept at room temperature was analyzed after 24 h. One of the samples kept at -20° was analyzed after one freeze-thaw cycle and the other was analyzed after one month storage. The peak area at the end of the study was compared with that of the standard and stability was calculated.For studying the pharmacokinetics of metformin by using the above developed method, metformin (320 mg/kg) was administered orallyIn the present method, acetonitrile was used to deproteinize plasma and simultaneously extract metformin. The recovery studies  indicate18 column using 0.15 M ammonium acetate (pH 5.5) and acetonitrile in the ratio of 90:10 as mobile phase, hled to a distinct curve for metformin at 4.7 min with good linearity indicating the preciseness and accuracy of the developed method. Most of the methods used potassium dihydrogen phosphate and acetonitrile in the mobile phase and used cyano or C8 column. However, our preliminary attempts revealed that use of ammonium acetate instead of potassium hydrogen phosphate in the mobile phase gives better resolution of metformin. This may be because metformin is a biguanide and possess amino groups and hence amino group possessing component in the mobile phase may be responsible for giving better resolution.When metformin extracted by acetonitrile from plasma was injected onto CIt was further noted that concentration of acetonitrile in mobile phase played a major role in the resolution of peak. At 10% concentration, the peak for metformin was optimally resolved with asymmetry of 0.9-1.4 and theoretical plates 18000-22000 tp/m. At higher concentration (>20%), the peak was broad whereas at lower concentration (< 10%), there was a tailing of peak which could be reduced by changing the pH of ammonium acetate buffer to 2.5. However, such acidic pH is not suitable for the life of the column. Therefore, 10% acetonitrile in the mobile phase was found more appropriate. In another experiment, methanol was used instead of acetonitrile for plasma preparation as well as in mobile phase. However, the ultimate peak height for metformin was very small and hence methanol was considered unsuitable.The typical chromatograms  obtained2 = 0.994). The inter-day and intra-day estimations of metformin revealed the reproducibility of the results irrespective of time and day. The calculated values of accuracy and precision  at 2.5±0.115 h (Tmax ) and AUC as 46.52±0.21 μg.h.ml-1 (The plasma level-time study of metformin in rats indicated highest metformin concentration in plasma to be 7.2±0.09 μg/ml (Cg.h.ml-1 . Seconda


The purpose of this study was to determine the prevalence of intestinal parasitic infections among patients who had attended Hawassa University students’ clinic, Southern Ethiopia.E. histolytica/E. dispar trophozoite was the most common identified parasite, which was seen in 18% of the patients while Enterobius vermicularis (0.1%) was the least reported parasite. Other parasites identified were Ascaris lumbricoides (15.0%), Hookworm species (2.0%), Taenia species (1.8%), Hymenolepis nana (0.7%), Strongyloides stercoralis (0.3%), Trichuris trichuria (0.2%), and Shistosoma mansoni (0.2%). The prevalence of helminthes was higher in females (23.3%) than in males (19.5%) (P < 0.00001) while that of protozoan infections was 28.5% in males than females (23.8%) (574/2414) (P < 0.00001).Over the 10 years period, a total of 13,679 patients visiting Hawassa University students’ clinic were included in the study. A total of 6553 (47.9%) patients were positive for at least one intestinal parasite. The overall prevalence of intestinal helminth and protozoan infections was 20.3% and 27.6% respectively. There were four dual infections and one triple infection.  Human intestinal parasitic infections are still the major causes of sickness and death henceforth, significant general medical issue globally . It is aIn sub-Saharan Africa, the prevalence of intestinal parasitic diseases is high, and its rate can extend up to 95%. In these counties up to 250 million individuals are assessed to be infected with something like at least one type of intestinal nematodes . These pThe study of disease transmission of intestinal parasitic infections demonstrates that these parasites are found in each age gathering and in both genders. Be that as it may, the rate is distinctive in certain areas and in some age groups . StudiesThis 10 years retrospective study was conducted in Hawassa city, Southern Ethiopia from August 2008 to July 2018.Hawassa city is located 273 km away from Addis Ababa capital of Ethiopia. The city has total population of 367,970. It is surrounded by Lake Hawassa. The city has an area of 157.2 SQ km.All Patients who had been examined for stool sample and having complete age, sex, and stool examination documentation over the study period.All patients’ data with complete age, sex, and stool examination documentation over the aforementioned study period were included in this retrospective study.Using data extraction sheet/form, all required information for this retrospective study was obtained from the registration/record books of the Hawassa University students’ clinic. After data collection process, the data was rechecked and cleaned.2) was used to determine association between prevalence by gender and age. P value was considered to be statistically significant when P value is less than 0.05. Additionally tables were used to display the results.All data were double analyzed using SPSS (version 23.0) statistical packages. Descriptive statistics were used for analysis. Chi square test , Ascaris lumbricoides (15.0%), and Giardia lamblia (9.6%). Enterobius vermicularis showed the least prevalence (0.1%). The other parasites identified were Hookworm Species, Trichuris trichiura, Schistosoma mansoni, Taenia species, Hymenolepis nana, Strongyloides stercoralis . The second dual IPIs was Ascaris lumbricoides and Entamoeba histolytica/dispar 79 (0.6%) following Hookworm species and Ascaris lumbricoides 49 (0.4%). The only triple IPIs was the combination of Entamoeba histolytica/dispar, Ascaris lumbricoides and Giardia lamblia 10 (0.1%)  Table .Table 2DHelminthic and protozoan infections specially are major health problem in Ethiopia. Many factors like, poor hygienic habits, poor standard of living, lack of health education, ignorance, poverty, poor socio-economic conditions are some of the many reasons behind high prevalence of parasitic infections. During the present study, protozoan infection was recorded to be higher compared to helminth infection. This condition usually contributed by different level of environmental conditions which facilitate the transmission of the infective stages of the parasites.The overall prevalence rate of intestinal parasitic diseases among patients record that had complete sex, age and stool examination at Hawassa University students’ clinic, from August 2008 to July 2018 was 47.9%. Such a rate of parasitic diseases recorded in this review study could for the most part be related to the low financial condition, described by deficient water supply, poor sterile transfer of excrement, the tropical atmosphere, and absence of information about parasite transmission common of many countries, for example, those in Africa ) [Entamoeba histolytica/dispar (18%) and Giardia lamblia (9.6%) in this study may be associated with poor personal and environmental hygiene in addition to having feco-oral route of transmission. This higher prevalence of these two protozoan parasites goes in line with the report of WHO which pointed out these two parasites as common causes of intestinal infection throughout Ethiopia [The predominant parasite identified in the present study was  (8.9%]) , 12. TheEthiopia .In this study the overall prevalence of intestinal helminth infections is 20.3% and it is significantly higher in females (23.3%) than males (19.5%). Higher prevalence in females (55.7%) than males (44.32%), was found a study conducted from Mwanza, Tanzania . But in The prevalence of protozoan infections was 27.6% and it was significantly higher in males (28.5%) than in females 23.8%) , and in age groups 24–33 years (48.5%) but lower in age groups 34+ years (39.6%). Age group specific prevalence of helminths for age group 18–23 years, 24–33 years and 34+ years were 20.3%, 20.1% and 18.9% respectively. Age group specific prevalence of protozoan for age group 18–23 years, 24–33 years and 34+ years were 27.7%, 28.4% and 20.7% respectively.Entamoeba histolytica/dispar was the most commonly reported parasite, which was seen in 18.0% of the patients. In conclusion, this study shows that intestinal helminthiases and protozoan infections are among the common parasitic infections observed among patients presenting at Hawassa University students’ clinic. It is necessary to develop effective prevention and control strategies including health education and improving environmental sanitation.The overall prevalence of intestinal parasitic infections in this retrospective study was 47.9% and was positive for at least one intestinal parasite. Entamoeba histolytica from Entamoeba dispar in stool samples was not possible.In this study, since microscopic examination method was used, differentiation of