Enhancement of ?-amylase production in pelleted Aspergillus tamarii through optimization for desizing of cotton fabric

Aim: The present study was carried out to enhance the production of α-amylase by pelleted Aspergillus tamarii through optimization of various media compositions and cultivation conditions using shake flask and bioreactor experiments. Methodology: Shake flask experiments were conducted to study the effect of pH, temperature and starch concentration using Response Surface Methodology (RSM) and other parameters, namely, nitrogen source, inoculum size and incubation days using single variable optimization technique for the pelleted growth of microorganism and amylase production. Scale up study was carried out for the assessment of results obtained from shake flask experiments using a laboratory scale bioreactor. In the bioreactor study, parameters, namely, pH control, agitation and aeration were considered. Results: Maximum amylase production using pelleted Aspergillus tamarii was achieved at initial pH 6.7, temperature 30.5 °C, 0.5% w/v starch, 0.1% w/v urea, 1.5% v/v inoculum size and 4 days of fermentation in the shake flask experiments. Filamentous growth was observed when the concentration of starch used was above 2%. The specific enzyme activity increased to 2.77 fold after partial purification. When enzyme was used for desizing cotton fabric, it produced 90% efficiency. The scale-up experiments revealed initial pH 6.7, agitation 300 rpm and aeration 1 vvm as the conditions suitable for pelleted growth, as well as to achieve maximum amylase production. Interpretation: The results indicate pelleted growth of Aspergillus tamarii and in turn achievement of maximum amylase activity depends on media composition and fermentation conditions used at the time of enzyme production. Efficient desizing of cotton fabric by amylase showed its effectiveness towards hydrolysis of starch and converting it to soluble products for easy removal.

Effects of intravenous dexmedetomidine on hyperbaric bupivacaine spinal anesthesia: a randomized study

The present study was designed to evaluate the effect of intravenous dexmedetomidine on spinal anesthesia with 0.5% of hyperbaric bupivacaine. One hundred American Society of Anesthesiologists [ASA] physical status I/II patients undergoing elective surgeries under spinal anesthesia were randomized into two groups of 50 each. Immediately after subarachnoid block with 3 ml of 0.5% hyperbaric bupivacaine, patients in group D received a loading dose of 1 microg/kg of dexmedetomidine intravenously by infusion pump over 10 min followed by a maintenance dose of 0.5 microg/kg/h till the end of surgery, whereas patients in group C received an equivalent quantity of normal saline. The time taken for regression of motor blockade to modified Bromage scale 0 was significantly prolonged in group D [220.7 +/- 16.5 min] compared to group C [131 +/- 10.5 min] [P < 0.001]. The level of sensory block was higher in group D [T 6.88 +/- 1.1] than group C [T 7.66 +/- 0.8] [P < 0.001]. The duration for two-dermatomal regression of sensory blockade [137.4 +/- 10.9 min vs. 102.8 +/- 14.8 min] and the duration of sensory block [269.8 +/- 20.7 min vs. 169.2 +/- 12.1 min] were significantly prolonged in group D compared to group C [P < 0.001]. Intraoperative Ramsay sedation scores were higher in group D [4.4 +/- 0.7] compared to group C [2 +/- 0.1] [P < 0.001]. Higher proportion of patients in group D had bradycardia [33% vs. 4%] [P < 0.001], as compared to group C. The 24-h mean analgesic requirement was less and the time to first request for postoperative analgesic was prolonged in group D than in group C [P < 0.001]. Intravenous dexmedetomidine significantly prolongs the duration of sensory and motor block of bupivacaine spinal anesthesia. The incidence of bradycardia is significantly higher when intravenous dexmedetomidine is used as an adjuvant to bupivacaine spinal anesthesia. Dexmedetomidine provides excellent intraoperative sedation and postoperative analgesia.

Respiratory gated simultaneous integrated boost-intensity modulated radiotherapy [SIB-IMRT] after breast conservative surgery for carcinoma of the breast: the Salmaniya medical complex experience

To present our clinical experience using SIB-IMRT Technique for Intact Breast cancer. A retrospective review of 45 cases of Stage I-IV breast cancer patients treated with SIB-IMRT with respiratory gating after conservative treatments from 35th November 2008 to 16th February 2010. The most common fractionation was 1.8 Gy to Ipsilateral Breast tissue and 2.2 Gy to the lumpectomy cavity giving whole breast dose as 50.4 Gy and Lumpectomy cavity dose as 50.4 Gy and Lumpectomy cavity dose as 61.6 Gy over 28 fractions concomitantly. Respiaratory gating was done and CT-images were taken in inspiration breath hold position. A total of 45 patients with breast cancer- stage [17.7%], II [71%], II [8.9%], IV [2.2%] were treated with SIB-IMRT with respiratory gated radiotherapy. Out of 45 patients, 24 are of left sided breast cancer and 21 are of right sided breast cancer patients. The median, Dose maximum [D-max] in SIB-IMRT is 106.2% of prescribed lumpectomy site dose. The median isodose line prescribed to PTV-2 is 100%. The Conformity index [CI] is 0.9688 [median value] and Homogeneity index [HII] 1.06 [median]. The median ipsilateral lung, mean dose is 21.66 Gy and V-20 is 37.4%. For left sided cases the median value of mean heart dose, V-30 and V-40 are 22.98 Gy, 23.45% and 9.45% respectively. Acute skin toxicity was of Grade-I in 2.2%, Grade-II in 64.4%, Grade-III in 31.1%, and Grade-IV in 2.2%. The global Breast cosmoses were seen excellent in majority [93%] of case at median follow up of 8 months duration. Breast SIB-IMRT Technique is feasible and comparable with other treatment techniques with reduced treatment duration by six fractions. At median follow up of 8 months the skin toxicity and cosmoses are excellent in high percentage of cases

Prognostic factors influencing pregnancy rate after stimulated intrauterine insemination

To determine the prognostic factors such as age, diagnosis, number of cycle attempts and semen parameters on the pregnancy rate of controlled ovarian hyperstimulation (COH) /intrauterine insemination (IUI). Three hundred and seventeen women who underwent 507 consecutive COH/IUI cycles were recruited from 1st January 2002 to 31st December 2005 inclusively. This retrospective study was done in University Malaya Medical Centre, a tertiary care academic centre. The main outcome measure was pregnancy rate according to age, infertility diagnosis, duration of infertility, semen parameters, and the number of treatment cycles. The overall pregnancy rates were 16.9% per cycle and 25.9% per couple. Pregnancy rates decreased with advancing maternal age. Pregnancy rate was also significantly lower in patient with postwash total motile sperm count (TMSC) 20 million/ml. The cumulative pregnancy rates varied greatly by diagnosis from 16% for patients with male factor infertility to 60% for patients with ovulatory disorder. Pregnancies among patients with male infertility, tubal factors infertility and endometriosis were achieved during the first three cycles. There is a clear age-related decline in fecundity associated with COH/IUI treatment. Women of > 40 years old, couple with postwash TMSC < or = 20 million/ml, severe endometriosis and tubal factors have a particularly poor prognosis.

A Comparative Study for Apoptosis on the Degree of the Amount of Photorefractive Ablation in Photorefractive Keratectomy

PURPOSE: This study was aimed to evaluate changes in the stromal keratocyte after ablation of 50 micrometer and 100 micrometer with use of photorefractive keratectomy(PRK). METHODS: At 4 hours, 24 hours, 72 hours, 7 days and 1 month after PRK, each group of rabbits including normal control group was treated with terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling(TUNEL) staining using ApopTag(R) kit in vivo, then apoptotic keratocytes were evaluated with light microscope. RESULTS: There was no response with TUNEL staining of the epithelial cells, stromal keratocyte, and endothelium in normal cornea. In the ablation group, however, regardless of the depth of photorefractive ablation, the TUNEL signal was maximal after 4 hours, and it decreased with time. The signal was more intense in 100 micrometer ablation group than 50 micrometer ablation group, although the signal was not observed at the endothelial cells in both groups. The number of apoptotic stromal keratocytes at each time point of 4 hr, 24 hr, 72 hr, and 1 week was 57+/-8.9, 49+/-7.5, 36+/-5.1, and 12+/-1.3 cells/field in 100 micrometer ablation group, and 31+/-4.4, 28+/-4.6, 21+/-3.9, and 5+/-1.1 cells/field in 50 micrometer ablation group, and the difference between the two groups was statistically significant(P<0.05). CONCLUSION: The more the amount of ablation with photorefractive keratectomy, the stronger the apoptotic response. The postoperative apoptotic response was observed especially within 1 week. These findings suggest that early suppression of postoperative apoptosis within 1 week will influence on the prognosis of visual quality after photorefractive keratectomy, and more studies will be needed in the future.

Confocal Microscopic and Histopathologic Changes in Corneal Morphology Under Corneal Scraped Injury in Rabbit

PURPOSE: To investigate that the effects of early wound healing stage after corneal epithelial scrape injury. METHODS: We studied the change of scraped corneal wound like corneal cells, corneal thickness, acelluar zone, and celluar morphology occurring at the time points of 1, 4, 12, 24, 48, 72 hours, and 7 days after corneal epithelial scrape injury by the confocal microscopy and EM findings in 4 each group rabbits. RESULTS: By normal confocal microscopy, the mean cell density was 891 cells/mm2 in the anterior stroma and decreased to 814 cells/mm2 in the middle stroma, 731 cells/mm2 in the posterior stroma, and the endothelial density was 3236 cells/mm2. The change in the morphology of the keratocyte nuclei from an elliptical shape anteriorly, to a more elongated shape posteriorly. Apoptosis revealed like as condensation or fragmentation of chromatin and nuclei, vesicle formation, apoptotic bodies after corneal scraped injury by EM findings. The mean thickness of normal cornea was as follow; 47 micrometer in the epithelium, 334 micrometer in the stroma, and 392 micrometer in total cornea. The thickness of postoperative cornea including stromal thickness and total thickness increased at the early wound healing stage, and then decreased to the postoperative 48 hours(P<0.001). Mean range of acellular zone in the stroma increased at the early wound healing, but significantly decreased at the postoperative 48 hours, 79 micrometer(P<0.001). CONCLUSIONS: Keratocyte cell density and corneal thickness at the three portions of cornea, the thickness of stromal acelluar zone, and the changes of cellular morphology were related with a kind of the early post-inflammatory reaction, especially 24 hours, of corneal scraped injury. It should be needed more studies concerned with control of early post-inflammatory reaction.