Ml Of Sterile Water Research Articles

PP-025 Treatment for suprachoroidal haemorrhage with intraocular alteplase: a case report

BackgroundThe intraocular inoculation of alteplase helps to dissolve blood clots.PurposeTo evaluate the effectiveness and safety of the use of alteplase in a patient with massive SCH who had to undergo...

Determination of enzymatic activities using a miniaturized system as a rapid method to assess soil quality

SummarySoil quality determination requires the analysis of a number of soil attributes using different approaches. In recent years, one of the most promising approaches has been the determination of enzymatic activities. Generally, only a few enzymes have been analysed and related to other soil properties such as total carbon, nitrogen content or microbial biomass carbon. The aim of this work was to investigate the possible use of the API ZYM strip, a semi‐quantitative miniaturized system that determines 19 enzymatic activities, to study soil quality. To this end, we tested the system in different soil types, including albic Arenosols, mollic Leptosols, rendzic Leptosols, haplic Leptosols and calcaric Regosols. Fresh samples were sieved through a 2‐mm sieve in the field and soil extracts were prepared by mixing 2–20 g (depending on the soil horizon characteristics) from each sample with 2–20 ml of sterile water. Next, 65‐µl aliquots of the supernatant extract were placed into each API ZYM microtube and were incubated at 37°C for 16 hours. Our results show important qualitative and quantitative differences among the different soil types studied, with soil characteristics and biological properties correlating with biochemical information. The results provide useful information not only to determine soil quality, but also to assess changes in the soil environment. As a whole, our results suggest that the use of the API ZYM system could prove most useful in soil environmental studies. Finally, some suggestions are presented, including modifications to the system that could improve its application in this field.

Micronucleated erythrocytes in newborn rats exposed to raltegravir placental transfer.

The use of raltegravir in treating HIV/AIDS has been proposed due to its effectiveness in suppressing high loads of HIV RNA in pregnant women, thus preventing infection of the fetus. However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring. The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes (MNE), micronucleated polychromatic erythrocytes (MNPCE), and polychromatic erythrocytes (PCE) in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir. The animals were randomly assigned to five groups as follows: raltegravir at doses of 15, 30, or 60 mg/day, cyclophosphamide 10 mg/kg (positive control), or 0.5 ml of sterile water (negative control). In addition, the effect of these drugs on the weight and height of newborns was assessed. There were no differences in the number of MNE, MNPCE, and PCE, and a slight decrease in the weight and height was observed in the offspring of the rat mothers treated with raltegravir. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses.

Studies on the Bacteria Burden in Garri Openly Sold in Port Harcourt Markets

A total of 30 samples of exposed garri in Port Harcourt (mile 1 Market, Churchill Market, Creek Road Market and control samples) were examined for burden of bacteria. One gram (1gm) of each garri was soaked in 9ml of sterile water to make a 1:10 dilution (stock solution) and 1ml was transferred to a test tube and diluted serially in one tenth stepwise to 10- 4 . From 10- 4 tube, an aliquot (0.1ml) of the diluted samples were inoculated on surface dried media yielding bacterial isolates. Colonies were counted to know the total viable bacterial count on the plates (expressed as colony forming unit/gram (Cfu/g) of garri). From the result obtained Mile 1 market had a mean standard deviation of 57.4 + 15.7 (white garri) and 51.6 + 21.5 (yellow garri), followed by Creek road market 46.6 + 10.2 (white garri) and 44 + 10.1 (yellow garri), then Churchill market 43.4+7.43 (white garri) and 41.8 + 9.2(yellow garri). The culture results showed growth of Staphylococcus aureus, 137 (49.3%), Staphylococcus epidermidis, 35 (12.6%), Bacillus sp., 59(21.2%), Escherichia coli, 43(15.5%), Proteus sp. 4(1.4%). The result of this study showed the presence of bacteria in gari sold in popular markets in Port Harcourt.

A selective medium for Xanthomonas axonopodis pv. Betlicola, bacterial pathogen of betelvine

Xanthomonas axonopodis pv. betlicola (Patel et al.) Vauterin et al. causes severe damage of betelvine (Piper betle L.) in West Bengal by producing different types of leaf spots (small to large, circular to irregular, angular), marginal leaf blight, stem lesion and wilting of vines. A selective medium was developed for isolation of this bacterium from diseased tissues and detection of this bacterium from the leaf surface, in soil and water. This bacterium grew best in Potato Sucrose Peptone Agar (PSPA) medium (Peeled potato 200 g, Sucrose 20 g, Peptone 5 g, Agar agar 20 g, and Water 1000 ml) and this medium was used as basal medium. Some fungicides and antibiotics (Carbendazim 25 mg, Copper oxychloride 25 mg, Metalaxyl 21 mg, Cycloheximide 50 mg, Pentid-200 100 mg, Nitrofurantoin 100 mg) were incorporated into the basal medium before use. Bavistin was used as source of Carbendazim, Blitox 50 as Copper oxychloride, Krilaxyl 35 WS as Metalaxyl, Furadantin capsule (human drug) as Nitrofurantoin. Pentid-200 is a human drug and it contains Penicillin-G potassium 2,00,000 units. The above fungicides and antibiotics were taken in a 25 ml sterilized conical flask plugged with cotton and 2 ml absolute alcohol was added to it. The flask was kept as such for 24 h to allow alcohol to evaporate. Then 20 ml of sterile water was added to the flask and shaken. The solution was kept in refrigerator for future use (30 days). The solution was mixed with the sterilized and melted PSPA medium prepared earlier at 4 ml/200 ml medium before use. Using this medium X. a. pv. betlicola was isolated successfully from the diseased tissue without surface sterilization. Key words: Bacterial disease, betelvine, selective medium, Xanthomonas axonopodis pv. Betlicola.

PRODUÇÃO, CARACTERIZAÇÃO E AVALIAÇÃO DE ENZIMAS FIBROLÍTICAS NA DIGESTIBILIDADE DA FORRAGEM DE MILHO

The objectives of this research were to produce and characterize an enzyme complex (EC), using the fungus Humicola grisea , and evaluate its effect on true digestibility of forage maize dry matter. We observed that the fungus produced cellulase, b-glucosidase and xylanase enzymes. The cellulase and xylanase activities were high at the temperature of 50° C. The optimum temperature of ?-glucosidase was between 50 and 60° C. The optimum pH of cellulase and xylanase enzyme was 6.0. As for ?-glucosidase, the enzyme showed higher activity at pH 6.5. Cellulase remained stable for 60 minutes at 39° C. Xylanase and ?-glucosidase maintained 99.2 and 88.2% of their activity at 50° C for 240 minutes, respectively. The treatments were as follows: control (10 mL of sterile water), level 1 (2.5 mL EC), level 2 (5.0 mL EC) and level 3 (10 mL EC). In the digestibility experiment, there was interaction between enzyme levels and time of incubation in the rumen. The addition of 10 mL of the fibrolytic enzymes improved the digestibility at 10.58; 12.52; 9.05 and 6.81% compared to control for 12; 24; 48 and 96 hours of incubation, respectively. The fungus Humicola grisea is an enzyme producer that is important in ruminant feed. KEYWORDS: Ankom; bovine; cellulose; fungi; xylanase.

Effect of aqueous extract of some plants on soil micro-fauna and micro-flora in vitro

Aqueous extract of five commonly available plant materials were evaluated for their effects on parasitic and free-living nematodes and fungi. Aqueous extract of Plumeria alba, Thevetia neriifolia, Jatropha curcas, Viscum album, Azadirachta indica and Hyptis suaveolens poit were prepared at two concentrations of 20 and 10%, with tap water as check. Parasitic nematodes (Helicotylenchus spp. and Scutellonema bradys and egg masses of Meloidogyne spp.) and free-living Rhabditis spp. were exposed to the extracts. Twenty grams each of plant materials were pounded and 20 ml of sterile water was added to make the extract. Each 400 ml of Potato Dextrose Agar + Streptomycin was amended with filtrate of an extract. A 2 mm plug of seven-day old mycelium block of Fusarium oxysporum f.sp. strigae and F. oxysporum, Telfairia isolate was inoculated each into an amended media. A media amended with 20 ml of sterile water served as check. The inhibitory effect of extract on fungus varied among the extract with P. alba and H. suaveolens poit having the greatest effect. All the extracts were lethal to parasitic and non-parasitic nematodes except J. curcas.

Evaluation of the presence of intraluminal cancer cells following rectal washout in rectal cancer surgery

The presence of intraluminal tumour cells in colorectal cancer is known, as is their ability to implant in colorectal anastomoses, causing perianastomotic tumour growth. There is no consensus as to the volume and agent to use for rectal washout. The purpose of this study was to assess the efficacy of our current routine for rectal washout in eliminating intraluminal tumour cells. Sixty consecutive patients undergoing low anterior resection or low Hartmann's procedure for rectal cancer at our unit were included. After the pelvic dissection was completed, the bowel was cross-clamped distal to the tumour and the prewashout sample (no. 1), was obtained by instilling 50 ml of saline and collecting the return fluid. A rectal washout followed using 500 ml of sterile water, 2 × 500 ml of 70 % ethanol and 500 ml of sterile water (sample no. 2-4) and the postwashout sample (no. 5) was obtained by using 50 ml of saline. The samples were examined and categorized as non-malignant or malignant at the Department of Pathology. The usual pathology report information was also assessed. In 33 of the 60 patients, cancer cells were identified in the prewashout sample. The rectal washout eliminated intraluminal cancer cells in 30 of the 33 patients. All of the remaining 3 had a higher blood loss (p = 0.026) and a tendency towards more polypoidal tumours (p = 0.053). A rectal washout volume larger than 2,000 ml might be necessary to eliminate all intraluminal tumour cells in total mesorectal excision for rectal cancer.

Simultaneous determination of methanol, acetaldehyde, acetone, and ethanol in human blood by gas chromatography with flame ionization detection

Methanol, acetaldehyde, acetone, and ethanol, which are commonly used as biomarkers of several diseases, in acute intoxications, and forensic settings, can be detected and quantified in biological fluids. Gas chromatography (GC)-mass spectrometry techniques are complex, require highly trained personnel and expensive materials. Gas chromatographic determinations of ethanol, methanol, and acetone have been reported in one study with suboptimal accuracy. Our objective was to improve the assessment of these compounds in human blood using GC with flame ionization detection. An amount of 50 µl of blood was diluted with 300 µl of sterile water, 40 µl of 10% sodium tungstate, and 20 µl of 1% sulphuric acid. After centrifugation, 1 µl of the supernatant was injected into the gas chromatograph. We used a dimethylpolysiloxane capillary column of 30 m × 0.25 mm × 0.25 µm. We observed linear correlations from 7.5 to 240 mg/l for methanol, acetaldehyde, and acetone and from 75 to 2400 mg/l for ethanol. Precision at concentrations 15, 60, and 120 mg/l for methanol, acetaldehyde, and acetone and 150, 600, and 1200 mg/ml for ethanol were 0.8-6.9%. Ranges of accuracy were 94.7-98.9% for methanol, 91.2-97.4% for acetaldehyde, 96.1-98.7% for acetone, and 105.5-111.6% for ethanol. Limits of detection were 0.80 mg/l for methanol, 0.61 mg/l for acetaldehyde, 0.58 mg/l for acetone, and 0.53 mg/l for ethanol. This method is suitable for routine clinical and forensic practices.

Modulation of Inflammatory Response in a Cirrhotic Rat Model with Induced Bacterial Peritonitis

Bacterial peritonitis is a severe complication in patients with cirrhosis and ascites and despite antibiotic treatment, the inflammatory response to infection may induce renal dysfunction leading to death. This investigation evaluated the effect of TNF-α blockade on the inflammatory response and mortality in cirrhotic rats with induced bacterial peritonitis treated or not with antibiotics. Sprague-Dawley rats with carbon-tetrachloride-induced cirrhosis were treated with an intraperitoneal injection of 109 CFU of Escherichia coli diluted in 20 mL of sterile water to induce bacterial peritonitis and randomized to receive subcutaneously-administered placebo, ceftriaxone, anti-TNF-α mAb and ceftriaxone, or anti-TNF-α mAb alone. No differences were observed between groups at baseline in respect to renal function, liver hepatic tests, serum levels of nitrite/nitrate and TNF-α. Treatment with ceftriaxone reduced mortality (73.3%) but differences did not reach statistical significance as compared to placebo. Mortality in rats treated with ceftriaxone and anti-TNF-α mAb was significantly lower than in animals receiving placebo (53% vs. 100%, p<0.01). Serum TNF-α decreased significantly in surviving rats treated with ceftriaxone plus anti-TNF-α mAb but not in treated with antibiotics alone. Additional studies including more animals are required to assess if the association of antibiotic therapy and TNF-α blockade might be a possible approach to reduce mortality in cirrhotic patients with bacterial peritonitis.

TCH-018 Formulation of an Oral Solution Containing “Potion JouliÈ” Phosphorus to Counteract the Shortage of Phosphoneuros

Background In October 2011 a shortage of Phosphoneuros became apparent. This oral solution containing phosphorus is prescribed in the treatment of diseases where phosphorus intake is essential. None of the...

The effect of the irrigant QMix on removal of canal wall smear layer: an ex vivo study

The aim of this study is to evaluate the effectiveness over application time of different formulations of a novel endodontic irrigant (QMix™ 2in1) composed of a polyaminocarboxylic acid chelating agent, a bisbiguanide antimicrobial agent, a surfactant and deionized water to remove the root canal smear layer and expose patent dentinal tubules compared to a standard solution of 17% EDTA. Eighty human tooth roots from extracted, single-rooted teeth were instrumented (size 40.06) using 0.2 mL of sodium hypochlorite (6.15%) between each file size with a 3 mL water rinse after final instrumentation. Eight groups of 10 roots were irrigated with 3 mL of different formulations of QMix: QMix A, QMix B, and QMix C, or 17% EDTA for 60 and 90 s, respectively, then rinsed with 5 mL of sterile water. The roots were irrigated using a standard irrigation syringe and a 30 ga side-vent needle with an apical-coronal motion to within 1 mm of the working length. The coronal, middle and apical thirds of one canal surface of each root was evaluated at 1000× using scanning electron microscopy. The presence of smear layer was scored using a 5-point scale. Data were analyzed with the Kruskal-Wallis rank sums test, the Steel-Dwass, all-pairs comparison test, and the Steel method (with control) test. Irrigant type was highly significant (p < 0.007). Combined 60 and 90 s exposure data indicated QMix A (p = 0.014) and QMix C (p = 0.028) were superior to EDTA. While at the 90 s exposure time, smear layer removal by solutions QMix A (p = 0.043), QMix B (p = 0.018), and QMix C (p = 0.011) was superior to EDTA. All irrigants removed smear layer more effectively at the coronal and middle levels compared to the apical level (p < 0.001). Analysis showed all three QMix formulations were superior to EDTA in smear layer removal and exposure of dentinal tubules in the root canal system in single-rooted teeth.

First Report of Marasmiellus palmivorus Causing Post-Emergence Damping Off on Coconut Seedlings in Malaysia.

In April and June 2010, coconut seedlings with symptoms of very slow growth, yellowing of leaves, and general abnormal leaf growth were observed in germination beds in Teluk Intan, Perak, Malaysia. The roots were soft, rotten, and brown, extending upward and downward from these lesions. Rhizomorphs and basidiocarps were produced on coconut seeds near the germination eye and identified as Marasmiellus palmivorus according description by Turner (2). Three isolates were obtained by plating surface sterilized symptomatic roots and basidiocarp on malt extract agar (MEA) amended with 85% lactic acid (1 ml added to 11 of the medium). To confirm the identity of the fungus, genomic DNA was extracted from mycelia and basidiocarps of isolates and the large subunit (LSU) region was amplified and sequenced using LR0R/LR7 primers (3). All isolates had identical LSU sequences (GenBank Accession No. JQ654233 to JQ654235). Sequences were identical to each other and 99% similar to a M. palmivorus sequence deposited in the NCBI database (Accession No. AY639434).To confirm pathogenicity, three isolates of M. palmivorus that were obtained from symptomatic plant tissue was inoculated onto seeds of Malaysian Red Dwarf variety. Each isolate was grown in 100 ml of malt extract broth in 250 ml Erlenmeyer flasks and incubated at 27 ± 2°C for 5 days on an orbital shaker (125 rpm). The resulting culture was passed through two layers of sterile cloth. Mycelial suspension was obtained by blending mycelia in 100 ml of sterile water. Seeds were sterilized by soaking in 10% v/v sodium hypochlorite in distilled water for 3 min. The seeds were then rinsed three times over running tap water. The calyx portion of the seed was removed and five holes were made around the germination eye. The seeds were inoculated by injecting 2 ml of suspension into each hole. The control seeds were inoculated with sterile distilled water only. The seeds were transferred to 40-cm diameter plastic pots containing a mixture of sand, soil, and peat in the ratio of 3:2:1, respectively, and steam treated at 100°C for 1.5 h. Pots were placed in the glasshouse with normal exposures to day-night cycles, temperatures of 29 ± 4°C, and high relative humidity (85 to 95%) achieved by spraying water twice daily. After 2 months, 75% of the inoculated seeds failed to germinate. It was speculated that the artificial inoculum was higher than under germination bed conditions. Rhizomorphs and basidiocarps were produced on husk seeds near the germination eye. Seedlings that emerged successfully developed symptoms similar to those observed in the germination bed. No symptoms developed in the noninoculated seeds and seedlings. At 80 days post inoculation, basidiocarps were observed emerging from three diseased seedlings near the germination eye. Three reisolations were made on MEA from root lesions surface sterilized. Pathogenicity tests and LSU sequence analyses indicated that M. palmivorus is the causal agent of the symptoms observed on coconut seedlings. M. palmivorus was first recorded on coconuts and oil palm in the 1920s (1) and attacks the fruit and the petiole on oil palm (2). To our knowledge, this is the first report of M. palmivorus causing post-emergence damping off on coconut seedlings.

First Report of Colletotrichum boninense Causing Anthracnose on Pepper in China.

Anthracnose, caused by Colletotrichum spp., is a severe disease and results in large losses in pepper (Capsicum frutescens) production in China (4). Colletotrichum boninense is one of the Colletotrichum species in pepper in China. In August 2011, anthracnose symptoms (circular, sunken lesions with orange to black spore masses) were observed on pepper fruits in De-Yang, Sichuan Province, China. Three single-spore isolates (SC-6-1, SC-6-2, SC-6-3) were obtained from the infected fruits. A 5-mm diameter plug was transferred to potato dextrose agar (PDA); the isolates formed colonies with white margins and circular, dull orange centers. The conidia were cylindrical, obtuse at both ends, and 10.5 to 12.6 × 4.1 to 5.0 μm. The colonies grew rapidly at 25 to 28°C, and the average colony diameter was 51 to 52 mm after 5 days on PDA at 25°C. Based upon these characters, the causal agent was identified as C. boninense. To confirm the identity of the isolates, the internal transcribed spacer (ITS) regions were amplified with the ITS1/ITS4 universal primers (1). The internal transcribed spacer (ITS) sequences (Accession No. JQ926743) of the causal fungus shared 99 to 100% homology with ITS sequences of C. boninense in GenBank (Accession Nos. FN566865 and EU822801). The identity of the causal agent as C. boninense was also confirmed by species-specific primers (Col1/ITS4) (2). In a pathogenicity test, five detached ripe pepper fruits were inoculated with 1 μl of a conidial suspension (106 conidia/mL) or five fruits with 1 μl of sterile water were kept as control. After 7 days in a moist chamber at 25°C, typical anthracnose symptoms had developed on the five inoculated fruits but not on control fruits. C. boninense was reisolated from the lesions, and which was confirmed by morphology and molecular methods as before. There have reports of C. boninense infecting many species of plants, including pepper (3). To our knowledge, this is the first report of C. boninense causing anthracnose on pepper in China.

Effects of Antioxidants Vitamin E and C on ErythrocyteFragility, HaemoglobinIndex and Colonic Temperature of Transported Japanese Quails (Coturnixcoturnix japonica)

The present study investigated the effects of antioxidants Vitamin E (VE) and C (ascorbic acid, AA) on erythrocytes osmotic fragility (EOF), haemoglobin index (HI), colonic temperature (CT) and recovery times of transported Japanese quails during the hot-dry conditions. Two hundred male Japanese quails were used. The birds were randomly and equally divided into four groups: VE and AA groups, administered with 0.5 mg/kg and 200 mg/ kg body weight of VE and AA, respectively, control transported group (control group), administered with 0.5 ml of sterile water, and sedentary non-transported group (S). One hour after the administration of the antioxidants, the birds were transported by road for 2 h. The EOF, HI and CT were recorded before transportation, immediately after transportation (0) and on 3rd and 7th day of post-transportation period. The results showed that transportation induced hyperthermia, increased EOF and hypochromic erythrocytes in the quails, the effects of which were mitigated by antioxidants VE and AA. Base-line values for EOF were returned to normal in the control, AA and VE quails on days 7, 3 and 0, respectively post-transportation. The findings showed that recovery time after transportation in quails should be 7 days; but the duration may be reduced to 3 days if treated with AA or eliminated completely if the quails are treated with VE before transportation. The ameliorating effect was more pronounced in quails administered with antioxidant VE. The EOF may be employed as a biomarker and additional diagnostic tool for the assessment of transport-induce oxidative stress in quails.

Usefulness of Contrast-Enhanced Endoscopic Sonography for Discriminating Mural Nodules From Mucous Clots in Intraductal Papillary Mucinous Neoplasms

The aim of this study was to evaluate the ability of contrast-enhanced endoscopic sonography for discrimination of mural nodules from mucous clots in intraductal papillary mucinous neoplasms of the pancreas. Contrast-enhanced endoscopic sonography was performed in 17 consecutive patients who had an intraductal papillary mucinous neoplasm with mural lesions. To perform contrast-enhanced endoscopic sonography, we used a second-generation sonographic contrast agent. After reconstitution with 2 mL of sterile water for injection, 0.7 mL of the agent was administered through a peripheral vein. From 10 to 30 seconds after injection of the contrast agent, the presence or absence of vascularity in mural lesions was assessed. All cases were referred to surgery, and diagnoses were finally obtained by pathologic examination. Diagnoses of mural nodules versus mucous clots based on the sonographic results were compared with tumor histopathologic findings. Pathologic findings revealed 12 cases with mural nodules and 5 cases without. Contrast-enhanced endoscopic sonography depicted vascularity in all 12 cases with pathologically confirmed mural nodules, whereas all 4 cases without vascularity had mucous clots. Only 1 case without a pathologically confirmed mural nodule was overestimated by contrast-enhanced endoscopic sonography as having a mural nodule. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of contrast-enhanced endoscopic sonography for mural nodule detection were 100%, 80%, 92%, 100%, and 94%, respectively. Evaluation of vascularity by contrast-enhanced endoscopic sonography could be useful for distinguishing mural nodules from mucous clots in intraductal papillary mucinous neoplasms. Contrast-enhanced endoscopic sonography could be a necessary option to determine surgical indications for intraductal papillary mucinous neoplasms when mural lesions are observed.

Pain reduction by continuous intraperitoneal nebulization of ropivacaine during gynecological laparoscopic surgery

The objective of this study was to examine the analgesic effect of intraperitoneal nebulization of ropivacaine during gynecological laparoscopic procedures for patients anesthetized with an ultrashort-acting opiate. The study was a double-blinded placebo-controlled randomized trial (Canadian Task Force classification I) and involved 40 patients (20 patients in each arm) undergoing elective gynecological same-day outpatient laparoscopic surgery including unilateral/bilateral salpingo-oophorectomy or unilateral/bilateral ovarian cystectomy at the University Hospital Ambulatory Gynecological Endoscopic Unit. The study group received 10 ml of 1 % ropivacaine, and the control group received 10 ml of sterile water by intraperitoneal nebulization. Vital signs were recorded and summarized. Postoperatively, patients were followed up for 24 h including visual analogue scale (VAS) scores and analgesic usage. Results showed no significant differences between the two groups in terms of patient characteristics and surgical data. There were no significant differences between the groups in postoperative VAS scores at rest at the different time intervals. VAS scores during strain showed significantly decreased pain at the postoperative 2-h interval (p < 0.05). Postoperative opiate and nonopiate analgesia consumption was similar for both groups. Patients in the study groups required more antiemetic agents compared to the control group patients (p < 0.05). There were no reported side effects related to ropivacaine. Intraperitoneal nebulization of 100 mg of ropivacaine during gynecological laparoscopy under general anesthesia with an ultrashort-acting opiate does not reduce postoperative pain and it does not reduce postoperative opiate and nonopiate analgesia consumption.

First Report of Target Spot Caused by Corynespora cassiicola on Cotton in Georgia

In 2005, crop consultants in southwestern Georgia reported an unusual occurrence of leaf spot in cotton (Gossypium hirsutum L.). Initial symptoms first developed as brick red dots that led to the formation of irregular to circular lesions with tan-to-light brown centers. Lesions further enlarged and often demonstrated a targetlike appearance formed from concentric rings within the spot. Observations included estimates of premature defoliation up to 70%, abundant characteristic spots on the leaves and bracts, and losses of several hundred kg of lint/ha. When symptomatic leaves were submitted to the University of Georgia Tifton Plant Disease Clinic in Tifton, GA, for identification in 2008, the causal agent was tentatively diagnosed as Corynespora cassiicola (Berk. & M.A. Curtis) C.T. Wei on the basis of similar symptoms and signs previously reported on cotton (3). In September 2011, symptomatic leaves were obtained from diseased cotton within a field (var. DP 1048B2RF) near Attapulgus, GA. Symptomatic tissue from diseased leaves was surface disinfested in 0.5% sodium hypochlorite for 1 min and plated on potato dextrose agar (PDA). Ten isolates were incubated at 21.1°C for 2 weeks with a 12/12 h light/dark cycle using fluorescent light located approximately 70 cm above the cultures. After 1 week, two isolates were transferred to quarter strength PDA for enhanced sporulation and were grown under the same conditions. Conidiophores from the isolated fungus were simple, erect, intermittently branching and septate, and gave rise to single, subhyaline conidia. Conidia had 4 to 17 pseudosepta and were 50 to 197 μm long and 7 to 16 μm wide, straight to curved, and obclavate to cylindrical. Pathogenicity tests were conducted by spraying 10 cotton seedlings (DP 555BR and DP 1048B2RF, two to four true leaf stage) until runoff with a blended suspension from a 2-week-old pure culture of the fungus diluted with 100 mL of sterile water. Five plants were sprayed with sterile water as noninoculated controls. Cotton seedlings were then incubated in a moist chamber at 21.1°C for 48 h. Within 1 week, all inoculated plants showed symptoms similar to those of diseased field plants. Symptoms were not observed on noninoculated control plants. The fungus was reisolated five times from symptomatic leaves and grown in pure culture. Conidia and conidiophores were identical to the morphology of the original isolates, and were similar to descriptions of C. cassiicola (2). To confirm the identity of the pathogen, DNA was extracted from a week-old culture and amplified with specific primers for loci "ga4" and "rDNA ITS" (1). DNA sequences obtained with the Applied Biosystems 3730xl 96-capillary DNA Analyzer showed 99% identity to C. cassiicola from BLAST analysis in GenBank. The resulting sequence was deposited into GenBank (Accession No. JQ717069). To our knowledge, this is the first report of this pathogen in Georgia. Given the increasing prevalence of this disease in southwestern Georgia, its confirmation is a significant step toward management recommendations for growers. Because foliar diseases caused by C. cassiicola are commonly referred to as "target spot" in other crops (e.g., soybeans), it is proposed that Corynespora leaf spot of cotton be known as "target spot of cotton."

First Report of Laurel Wilt Disease Caused by Raffaelea lauricola on Silk Bay in Florida.

Silk bay (Persea humilis Nash) is a member of the Lauraceae precinctive to the scrub forests of central and southern Florida and a sister species to the primary laurel wilt host, redbay (P. borbonia (L.) Spreng), which is generally not found in these ecosystems. In November 2011, observations of silk bay mortality near Lake Placid in Highlands County, FL, were reported to Florida Forest Service agents. A subsequent visit to roadside and homeowners' properties in the area revealed many dead and dying silkbays with characteristic laurel wilt symptoms, including wilted crowns with brown persistent foliage, frass accumulated at tree bases, sapwood with dark streaking, and ambrosia beetle entrance holes (1). Twig samples were taken and later confirmed as silk bay by the Florida Division of Plant Industry in Gainesville. Trunk samples were taken from four trees for fungal isolation. Stem sections with vascular discoloration were surface disinfested for 30 s in a 5% sodium hypochlorite solution and then plated onto cycloheximide streptomycin malt extract agar (CSMA) (1). All sapwood pieces from the four samples resulted in the same cream-buff submerged fungal growth characteristic of the laurel wilt pathogen Raffaelea lauricola T. C. Harr., Fraedrich & Aghayeva (2). DNA was extracted from a single-spore derived isolate, PL1389, and an 18S small subunit ribosomal RNA gene sequence was generated with primers NS1 and NS4, resulting in a 1,031-bp amplicon (3). A BLASTn search showed identical homology to R. lauricola strains PL159 and PL382 (GenBank Accessions No. EU257806 and JF797171, respectively, 100% similarity, e-value 0.0, and a total score of 1,982). The sequence was deposited into GenBank and assigned the accession No. JQ247569. In December 2011, a spore suspension was made by flooding a PL1389 culture plate with 2 ml of sterile water, collecting by pipette, and quantification and adjusting to 3.25 × 106 spores/ml by hemacytometer. Pathogenicity was tested on potted plants in a growth chamber experiment. Five silk bays and three redbays were drill-wounded with a 3/32" drill bit and inoculated with 20 μl of the spore suspension. Three silk bays and two redbays served as water-inoculated controls. Within 5 weeks, all inoculated plants displayed the wilt and vascular discoloration characteristic of laurel wilt disease, while all water-inoculated controls remained healthy. Sapwood samples from all plants were plated onto the same CSMA media. R. lauricola was later recovered only from the wilted plants inoculated with PL1389, while no fungal growth was recovered from the asymptomatic water-inoculated controls. Silk bay, which plays a significant role in the limited scrub ecosystems of Florida, has now become another host in the laurel wilt epidemic, with its implications upon the scrub forests yet to be seen.

Comparative evaluation of apical extrusion of debris and irrigant with three rotary instruments using crown down technique – An in vitro study

Aims and objectiveThe purpose of this study was to evaluate and compare the apical extrusion of debris and irrigant using various rotary instruments with crown down technique in the instrumentation of root canals. Material and MethodThirty freshly extracted human permanent straight rooted mandibular premolars with minimum root curvature of 0–10 ° were divided in three groups with 10 teeth in each group. Each group was instrumented using one of the three rotary instrumentation systems: Rotary Hero shapers, Rotary ProTaper and Rotary Mtwo. One ml of sterile water was used as an irrigant after using each instrument. Debris extruded was collected in pre weighed glass vials and the extruded irrigant was measured quantitatively by Myers and Montgomery method and was later evaporated. The weight of the dry extruded debris was calculated by comparing the pre and post instrumentation weight of glass vials for each group. Statistical analysisStatistical analysis was done by using by a Kruskal–Wallis One-way ANOVA test. ResultsStatistical analysis showed that all the rotary instruments used in this study caused apical extrusion of debris and irrigant. A Statistically significant difference was observed with Rotary ProTaper and Rotary Mtwo groups when compared with Rotary Hero shapers. But no significant difference was observed between Rotary ProTaper and Rotary Mtwo groups. ConclusionAfter instrumentation with different rotary instruments, Hero shapers showed a less apical extrusion of debris and irrigant.