Investigation of selective JAK1 inhibitor GSK2586184 for the treatment of psoriasis in a randomized placebo-controlled phase IIa study.

BACKGROUND: GSK2586184 is a selective oral Janus kinase (JAK)1 inhibitor being evaluated as a treatment for moderate-to-severe plaque-type psoriasis. OBJECTIVES: To assess the relationship between dose of GSK2586184 and clinical response, primarily by the Psoriasis Area Severity Index (PASI). METHODS: Sixty patients with moderate-to-severe plaque psoriasis were randomized to cohort A: 100 mg, 200 mg or 400 mg GSK2586184 twice daily or placebo; and eight were randomized to open-label cohort B, a small exploratory cohort treated with 400 mg GSK2586184 twice daily, to explore differential gene expression. RESULTS: At week 12, a 75% reduction in PASI (PASI 75) response rates in the intent-to-treat population were 0% in the placebo group compared with 13%, 25% and 57% in the 100 mg, 200 mg and 400 mg GSK2586184 twice-daily groups, respectively. Increases in the proportion of PASI 75 responses were seen across all dose levels by week 4. Improvement in itch and quality of life were observed at all doses relative to placebo with the greatest improvement seen in the 400-mg dose group. Overall, the incidence of adverse events (AEs) was similar across treatment groups, and no relationship between frequency of AE and GSK2586184 dose was identified. Differential gene expression was observed in involved and uninvolved skin at baseline and in involved skin after 2 weeks of treatment with GSK2586184. CONCLUSIONS: Our study demonstrates that 12 weeks of treatment with GSK2586184 resulted in clinical improvement and was generally well tolerated in patients with moderate-to-severe plaque-type psoriasis.

Molecular characterization and detection of mutations associated with resistance to succinate dehydrogenase-inhibiting fungicides in Alternaria solani.

Early blight, caused by Alternaria solani, is an economically important foliar disease of potato in several production areas of the United States. Few potato cultivars possess resistance to early blight; therefore, the application of fungicides is the primary means of achieving disease control. Previous work in our laboratory reported resistance to the succinate dehydrogenase-inhibiting (SDHI) fungicide boscalid in this plant pathogen with a concomitant loss of disease control. Two phenotypes were detected, one in which A. solani isolates were moderately resistant to boscalid, the other in which isolates were highly resistant to the fungicide. Resistance in other fungal plant pathogens to SDHI fungicides is known to occur due to amino acid exchanges in the soluble subunit succinate dehydrogenase B (SdhB), C (SdhC), and D (SdhD) proteins. In this study, the AsSdhB, AsSdhC, and AsSdhD genes were analyzed and compared in sensitive (50% effective concentration [EC50] < 5 µg ml(-1)), moderately resistant (EC50 = 5.1 to 20 µg ml(-1)), highly resistant (EC50 = 20.1 to 100 µg ml(-1)), and very highly resistant (EC50 > 100 µg ml(-1)) A. solani isolates. In total, five mutations were detected, two in each of the AsSdhB and AsSdhD genes and one in the AsSdhC gene. The sequencing of AsSdhB elucidated point mutations cytosine (C) to thymine (T) at nucleotide 990 and adenine (A) to guanine (G) at nucleotide 991, leading to an exchange from histidine to tyrosine (H278Y) or arginine (H278R), respectively, at codon 278. The H278R exchange was detected in 4 of 10 A. solani isolates moderately resistant to boscalid, exhibiting EC50 values of 6 to 8 µg ml(-1). Further genetic analysis also confirmed this mutation in isolates with high and very high EC50 values for boscalid of 28 to 500 µg ml(-1). Subsequent sequencing of AsSdhC and AsSdhD genes confirmed the presence of additional mutations from A to G at nucleotide position 490 in AsSdhC and at nucleotide position 398 in the AsSdhD, conferring H134R and H133R exchanges in AsSdhC and AsSdhD, respectively. The H134R exchange in AsSdhC was observed in A. solani isolates with sensitive, moderate, highly resistant, and very highly resistant boscalid phenotypes, and the AsSdhD H133R exchange was observed in isolates with both moderate and very high EC50 value boscalid phenotypes. Detection and differentiation of point mutations in AsSdhB resulting in H278R and H278Y exchanges in the AsSdhB subunit were facilitated by the development of a mismatch amplification mutation assay. Detection of these two mutations in boscalid-resistant isolates, in addition to mutations in AsSdhC and AsSdhD resulting in an H134R and H133R exchange, respectively, was achieved by the development of a multiplex polymerase chain reaction to detect and differentiate the sensitive and resistant isolates based on the single-nucleotide polymorphisms present in all three genes. A single A. solani isolate with resistance to boscalid did not contain any of the above-mentioned exchanges but did contain a substitution of aspartate to glutamic acid at amino acid position 123 (D123E) in the AsSdhD subunit. Among A. solani isolates possessing resistance to boscalid, point mutations in AsSdhB were more frequently detected than mutations in genes coding for any other subunit.

First Report of Colletotrichum higginsianum Causing Anthracnose of Arugula (Eruca sativa) in Florida.

Arugula (Eruca sativa) is grown in Florida and is an important component in packaged salad products. During spring 2013, leaf lesions on arugula caused significant economic losses in Miami-Dade County, Florida. Symptoms initially appeared as small water-soaked lesions that later became circular, sunken, and white in the center with a dark brown to black halo, up to 4 mm in diameter. Acervuli were found under a dissecting microscope on infected leaf lesions with black spines or setae. Occasionally, small, circular, often longitudinal dark brown spots appeared on leaf branches. Leaf tissues (5 × 5 mm) from lesion margins were surface sterilized in 0.9% sodium hypochlorite for 10 min, rinsed with sterile distilled water, and plated on potato dextrose agar (PDA). PDA plates were incubated at 21°C under 24-h fluorescent lights for 4 to 6 days. The fungus initially produced gray mycelium followed by orange conidial mass. Hyphae of the fungus were septate and hyaline. After 5 to 7 days, the fungus produced acervuli with dark brown to black setae (75 to 130 µm long) (n = 20). Conidia were found in the colonies, which were single celled, oblong, hyaline, and 12 to 25 × 4 to 6 µm (n = 20). The cultural and morphological characteristics of the conidia were similar to those for Colletotrichum higginsianum Sacc (1). To further confirm the species of the isolates, the sequence of the ITS region of rDNA, chitin synthase 1 (CHS1), and actin (ACT) was amplified from isolates 05131 and 05132 using primer pairs ITS 1 and ITS 4 (4), CHS-79F and CHS-354R, and ACT-512F and ACT-783R (3), respectively. The sequenced data of each locus were deposited in GenBank with accessions KF550281.1, KF550282.1, KJ159904, KJ159905, KJ159906, and KJ159907. The resulting sequence of ITS showed 100% identity with sequences of C. higginsianum in JQ005760.1, and sequence of ACT gene showed 100% identity with C. higginsianum in JQ005823.1. The sequence of ACT gene and ITS region had ≤99% identity with other closely related Colletotrichum spp. CHS1 gene had 100% identity with JQ005781.1 belonging to C. higginsianum, and one accession JQ005783.1 belonging to C. fuscum. However, ACT gene and ITS region does not share 100% identity with C. fuscum and therefore, sequence data from three loci proves that isolated pathogen is C. higginsianum. All the above mentioned accessions that shared 100% identity with sequences of isolates used in our study have been previously used to represent the species in the C. destructivum clade in a systematics study (2). To confirm its pathogenicity, a suspension of isolate 05132 at 5 × 105 conidia/ml was sprayed on leaves of five arugula plants until runoff. The other five arugula plants sprayed with water served as non-inoculated controls. Both inoculated and non-inoculated plants were separately covered with a plastic bag to maintain high humidity for 24 h at 27 ± 5°C under natural day/night conditions in the greenhouse. Symptoms first appeared 3 to 4 days after inoculation as small water-soaked lesions, which became sunken with dark brown to black margins. Small circular and longitudinal dark brown spots were also seen on leaf branches as seen initially on naturally infected arugula. No symptoms developed on non-inoculated control plants. C. higginsianum was re-isolated from the lesions with the same morphological characteristics as described above, fulfilling Koch's postulates. To our knowledge, this is the first report of C. higginsianum causing anthracnose of arugula in Florida. This pathogen may potentially affect the salad industry in the United States. References: (1) A. J. Caesar et al. Plant Dis. 94:1166, 2010. (2) P. F. Cannon et al. Stud. Mycol. 73:181, 2012. (3) I. Carbone and L. M. Kohn. Mycologia 91:553, 1999. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Elm Canker Caused by Pestalotiopsis mangiferae in the United States.

Chinese elm (Ulmus parvifolia Jacq.) is native to China, Korea, and Japan and was introduced to the U.S. to replace the American elm, which is highly susceptible to Dutch elm disease. Cultivar Emmer II trademark Allee elm (or Allee elm) is preferred by nurseries for its rich green foliage and beauty of bark. In the summer of 2011, a new disease was observed on Allee elm at a tree farm in FL. Approximately 1% of elms in the same farm and in residential areas in central Florida had similar canker-like symptoms consisting of tan to orange patches of decomposed and loose bark. These symptoms were observed on the main trunks often extending into branches of affected trees. Cankered sections of the trunk were often several feet in length and penetrated the wood to a depth just under the bark into the phloem. To isolate the causal organism, cankers were gathered from 7 trees and tissues from the margin of 1 to 3 cankers per tree were surface sterilized in 1% sodium hypochlorite, plated on PDA, and incubated at 22°C under a 16-h/8-h light/dark cycle for 7 days. Colonies displayed white, fluffy mycelium with sporadic black acervuli containing aggregated conidia. Conidia were 5-celled with two or more apical appendages or hairs; the three central cells were dark brown and the two outer cells were hyaline (1). Based on conidial morphology, these isolates were putatively identified as Pestalotiopsis spp. To identify the species, the rDNA internal transcribed spacer (ITS) region was sequenced for two field isolates (GenBank Accessions JX999998 and JX999999). A BLASTn search in GenBank revealed 100% identity to Pestalotiopsis mangiferae ITS (JX305704.1). To test Koch's postulates, experiments were performed in the field and greenhouse. A mycelial plug of isolate 11-40 was grown on PDA, inoculated on wounded trunk of healthy 18-month-old Allee elms (n = 48) in the field and 7-month-old Allee elms (n = 12) in the greenhouse. Both experiments were set up as a randomized complete block design. The trunk of each tree was wounded with a scalpel to a depth of 5 mm, the wound was inoculated with a 5-mm2 agar plug from a 7-day PDA culture, and the inoculated wound was wrapped with grafting tape. Plants that served as negative controls [n = 20 (field experiment) and n = 12 (greenhouse)] were mock-inoculated with a sterile PDA plug without mycelial growth. After 4 to 6 months, symptoms consisting of loose or fallen off bark developed on all pathogen-inoculated trees but not on control trees. Control wounds healed with no expansion beyond the original 5 mm inoculation zone, whereas Pestalotiopsis-inoculated cankers expanded to 3 to 8 cm in each direction in 6 months. Pestalotiopsis (confirmed by conidial morphology and ITS sequencing) was reisolated from pathogen-inoculated trees but not from control trees. These experiments were repeated with similar results at least three times, each consisting of 15 replications (greenhouse) and 12 replications (field) with additional Pestalotiopsis isolates. Many other Pestaliopsis spp. have been reported on other shade trees through the U.S. and the world. To our knowledge, this is the first report of P. mangiferae on an Ulmus species in the U.S. and the world. Since elm canker mainly affects the trunk, a featured characteristic of the Allee elm, it can potentially result in economic loss to the ornamental industry. References: (1) Y. Ko et al. Plant Dis. 91:1684, 2007. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Body temperature around induced estrus in dairy cows.

The overall objective of this study was to study the influence of induced estrus on body temperature, comparing 5 distinct intervals around induced estrus and to determine the diurnal pattern from 4 ± 1 d before to 4 ± 1 d after induced estrus. Sixteen estrous cycles of 9 postpartum dairy cows were synchronized with 2 injections of PGF(2α), 10 d apart. After the second PGF(2α) injection on d 10, temperature loggers were inserted into the vaginal cavity for a 12 ± 1-d period. Two days later, a third dose of PGF(2α) was injected to induce estrus. After confirmation of a corpus luteum, loggers were removed on d 5 ± 1. Observation of estrus, rectal palpation, and ultrasound scanning to determine ovulation were carried out every 4 ± 1h, beginning at 12h after the third PGF(2α) injection. Blood samples from the vena coccygea mediana were collected twice daily from d 11 to 12 and every 4 ± 1h after the third PGF(2α) injection until ovulation. Vaginal temperature was recorded every 5 min and averaged to hourly means for the following 5 periods: 1) 48 h preceding the third PGF(2α) injection, 2) from the third PGF(2α) injection to first signs of estrus, 3) estrus to ovulation, 4) a 4-h interval in which ovulation occurred, and 5) a 96-h post-ovulation period. High body temperatures (39.0 ± 0.5 °C) and low progesterone (P4) concentrations (<0.5 ng/mL) were observed during estrus, whereas low body temperatures were observed from PGF(2α) injection to estrus (38.6 ± 0.3 °C) and around ovulation (38.5 ± 0.2 °C), respectively. An association between body temperature and serum P4 concentrations did not exist. However, P4 concentrations on d 11 and 12 were high (5.0 ± 1.5 ng/mL) and decreased (0.9 ± 0.2 ng/mL) after ovulation. Diurnal temperature rhythms were similar before and after estrus. Vaginal temperature before estrus (d 11 and 12) was slightly (0.1 °C) higher compared with the post-ovulation period.

Longevity of Crown Margin Repairs Using Glass Ionomer Cement: A Retrospective Study.

OBJECTIVE: The objective of this study was to determine the survival time of crown margin repairs (CMRs) with glass ionomer and resin-modified glass ionomer cements on permanent teeth using electronic dental record (EDR) data. METHODS: We queried a database of EDR (axiUm; Exan Group, Coquitlam, BC, Canada) in the Indiana University School of Dentistry (IUSD), Indianapolis, IN, USA, for records of patients who underwent CMRs of permanent teeth at the Graduate Operative Dentistry Clinic. Two examiners developed guidelines for reviewing the records and manually reviewed the clinical notes of patient records to confirm for CMRs. Only records that were confirmed with the presence of CMRs were retained in the final dataset for survival analysis. Survival time was calculated by Kaplan-Meier statistics, and a Cox proportional hazards model was performed to assess the influence of age, gender, and tooth type on survival time (a<0.05). RESULTS: A total of 214 teeth (115 patients) with CMR were evaluated. Patient average age was 69.4 ± 11.7 years old. Posterior teeth accounted for 78.5% (n=168) of teeth treated. CMRs using glass ionomer cements had a 5-year survival rate of 62.9% and an annual failure rate (AFR) of 8.9%. Cox proportional-hazards model revealed that none of the factors examined (age, gender, tooth type) affected time to failure. CONCLUSION: The results indicate the potential of CMRs for extending the functional life of crowns with defective margins, thus reducing provider and patient burden of replacing an indirect restoration. We recommend future studies with a larger population who received CMR to extend the generalizability of our findings and to determine the influence of factors such as caries risk and severity of defects on survival time.

A plant biostimulant made from the marine brown algae Ascophyllum nodosum and chitosan reduce Fusarium head blight and mycotoxin contamination in wheat.

Fusarium head blight (FHB) caused by Fusarium graminearum is a disease that results in yield loss and mycotoxin contamination in wheat globally. This study assessed the effect of a plant biostimulant prepared from a brown macroalga Ascophyllum nodosum (Liquid Seaweed Extract; LSE) alone and in combination with chitosan in controlling Fusarium. Wheat seedlings drenched with LSE and chitosan in combination showed reduced severity of F. graminearum infection on leaves as evidenced by a significant reduction in necrotic area and fewer number of conidia produced in the necrotic area. Gene expression studies showed that the combination of LSE and chitosan amplified the response of pathogenesis-related genes (TaPR1.1, TaPR2, TaPR3, TaGlu2) in wheat seedlings infected with Fusarium spores above that observed for the individual treatments. The combination treatments were more effective in enhancing the activity of various defense related enzymes such as peroxidase and polyphenol oxidase. FHB studies on adult plants showed a reduction of bleached spikes in wheat heads treated with the combination of LSE and chitosan. Mycotoxin content appeared to be correlated with FHB severity. Combination treatments of LSE and chitosan reduced the levels of mycotoxins deoxynivalenol and sambucinol in wheat grains. Systemic disease resistance appears to be induced by LSE and chitosan in response to F. graminearum in wheat by inducing defense genes and enzymes.

Mechanism of vasorelaxant activity of a fraction of root extract of Sesamum indicum Linn.

The petroleum ether soluble fraction (SIPE) of the root extract of S. indicum was evaluated for the vasorelaxant activity using isolated rat aorta. SIPE up to 180 microg/ml concentration significantly inhibited phenylephrine- and KCl-induced contraction to the extent of 98.13 +/- 6.37 and 70.19 +/- 3.43% respectively in isolated rat aorta in a concentration dependent manner. The vasorelaxant activity was not blocked by propranolol (10 microM), atropine (1 microM) indomethacin (10 microM) and glibenclamide (10 microM). Influence of SIPE on phenylephrine-induced contractions in aortic preparations in absence of functional endothelium and on pre-incubating the tissue with L-NAME (300 microM) or methylene blue (10 microM) was also studied. SIPE at 180 microg/ml concentration could elicit partial relaxation in presence of L-NAME or methylene blue to the extent of 34.26 +/- 6.13 and 25.66 +/- 10.95% respectively. However, in absence of functional endothelium, SIPE exhibited little relaxation to the extent of 6.70 +/- 4.87%. These studies revealed that the vasorelaxant activity of SIPE was chiefly mediated through endothelium-dependent pathway.

Seasonal incidence of parasitic diseases in bovines of south western Gujarat (Junagadh), India.

The current study was conducted to investigate the incidence of parasitic diseases in bovines which were sick and brought at veterinary hospital for treatment. A total of 366 samples were investigated from cattle (n = 175) and buffaloes (n = 191) presented at Teaching Veterinary Clinical Complex (TVCC), Veterinary College, Junagadh, Gujarat during January to December 2014. Examination of Giemsa-stained peripheral blood smears exhibited that 58.6 % of cattle and 41.2 % of buffaloes were infected with haemoparasites comprising Babesia bigemina, Theileria annulata, and Anaplasma marginale @ of 54.0, 3.4 and 1.1 in cattle and 38.8, 1.2 and 1.2 percent in buffaloes, respectively. The incidence of total haemoparasites and B. bigemina infections was significantly higher (p < 0.05) in cattle whereas, incidence of haemoparasites were recorded significantly higher in the month of July to November (i.e., rainy and autumn) in both cattle and buffaloes, respectively (p < 0.01 and p < 0.001). Coprological examination revealed that the overall incidence of gastrointestinal (GI) parasitic infection was 45.5 % in cattle and 43.4 % in buffaloes. The incidence of individual parasite was 11.4, 1.1, 2.3, 4.5, 1.1, 3.4, 2.3 and 19.3 in cattle and 4.7, 0.9, 0.0, 2.8, 0.9, 5.7, 0.0 and 28.3 % in buffaloes for Eimeria spp., Trichuris spp., Toxocara vitulorum, Strongyle, Fasciola spp., amphistomes, Schistosoma indicum and Buxtonella sulcata, respectively which differ insignificantly (p > 0.05). Seasonal prevalence of GI parasites was highest in summer in both cattle and buffaloes, the data being statistically non-significant (p > 0.05). However, the incidence of B. sulcata in both cattle (19.3 %) and buffaloes (28.3 %) was higher in comparisons to other GI parasites. The present investigation emphasized that B. bigemina and B. sulcata are the most important parasites of bovines of this region.

Enantioselective α-Amination Enabled by a BINAM-Derived Phase-Transfer Catalyst.

Chiral anion phase-transfer of aryldiazonium cations was utilized to achieve highly enantioselective α-amination of carbonyl compounds. A broad scope of indanone- and benzosuberone-derived substrates was amenable to this strategy. Critical to obtaining high levels of enantioselectivity was the use of BINAM-derived phosphoric acids. The utility of this transformation was demonstrated through facile conversion of diazene products to valuable α-amino acid derivatives.

Spontaneous corneal rupture in Noonan syndrome. A case report.

PURPOSE/METHODS: A variety of ocular and periocular manifestations have been described in Noonan syndrome. Collagen abnormalities have been described; however, to our knowledge spontaneous corneal rupture has not been reported. A forty-three-year-old female who presented with spontaneous corneal rupture was later diagnosed as having Noonan syndrome. RESULTS/CONCLUSIONS: Collagen abnormalities have been described with Noonan syndrome and this is likely the cause of corneal rupture in this patient.

Distant and delayed mitomycin C extravasation.

Mitomycin C is a vesicant chemotherapeutic agent used to treat solid tumors. Its ability to cause delayed and remote tissue injury after intravenous administration is reported in the literature. Two cases of delayed and distant mitomycin C extravasation injury occurred in our institution. In both patients, no evidence of acute extravasation was visible during or immediately after administration. Within 48 hours, one patient reported erythema, burning, and pain in the hand contralateral to the administration site. The second patient developed three distinct ulcerated lesions on her forearm within 6 weeks of receiving the agent. The lesions, located at sites of venipunctures, occurred 12-15 cm proximal to the site of mitomycin infusion. Because the drug has the potential to cause such unusual and unanticipated tissue injuries, health care professionals and patients should be aware of this hazard.

Epikeratophakia to correct traumatic aphakia after penetrating keratoplasty.

A 19-year-old man with keratoconus sustained ocular trauma and became aphakic in his operated left eye 2 months after penetrating keratoplasty. Original corneal wound repair was performed without intraocular lens implantation. Attempts to correct his aphakia with a contact lens failed when the patient became intolerant to its use. As an alternative, the patient had elective epikeratophakia. A standard 8.5 mm lenticule was placed over existing corneal graft. This operation resulted in +12.25 diopters of correction and a best corrected visual acuity of 20/25 at 30 months postoperatively. There was no sign of abnormalities at the host cornea or the transplanted lenticule. This case indicates that epikeratophakia may be successfully performed over existing corneal grafts.

Levels of dichlorodiphenyltrichloroethane and hexachlorocyclohexane in human adipose tissue of the Indian population.

Concentrations of dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexane (HCH) were determined in 313 human omental fat samples collected from subjects from all five zones of India during 1977-1980. The median concentration of 2,2-bis-(p-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE), total DDT, beta HCH, and total HCH were 3.4, 6.0, 1.3, and 1.9 mg/kg, respectively. The calculated national mean levels for DDT and HCH were 11.1 and 3.5 mg/kg, respectively. Although the values of DDT and HCH were not as high as those reported earlier, there is still a need for close monitoring of the bioaccumulation of these chemical residues in the Indian population.

Use of a phosphorus-enriched dialysate to hemodialyze patients with ethylene glycol intoxication.

A phosphorus-enriched dialysate was used to intensively hemodialyze two non-uremic and non-hyperphosphatemic patients poisoned with ethylene glycol. As a consequence of the use of such a dialysate, we were able to prevent a fall in plasma inorganic phosphorus levels in these patients in spite of the intensive dialytic therapy.

Focal dermal hypoplasia (Goltz syndrome) presenting as a severe fetal malformation syndrome.

A fetal malformation syndrome comprising growth retardation, anophthalmia, bilateral diaphragmatic herniae, bifid lower leg, syndactyly of the fingers, malrotation of the colon, hypoplastic kidneys and total anomalous pulmonary venous drainage is described in a female fetus from a consanguineous relationship. Differential diagnosis is discussed and it is suggested that this case represents an unusually severe form of Goltz syndrome.

Benzidine and its acetylated metabolites in the urine of workers exposed to Direct Black 38.

Urine samples of workers in a small-scale unit manufacturing Direct Black 38 were analyzed by high-pressure liquid chromatography for the presence of benzidine and mono and diacetyl benzidine. Acetylated metabolites were found in all the urine samples, and benzidine was found in all except two. Two workers excreted very high levels of benzidine and its metabolites in their urine. This study highlights the potential risk of bladder cancer in such units of developing countries where manufacture of benzidine-based dyes is yet to be regulated.

Removal of a corneal foreign body through a lamellar corneal pocket.

A piece of glass was found in the posterior central cornea of a 43-year-old victim of a motor vehicle accident. The original entry of the foreign body had healed and epithelialized. This foreign body was removed through a peripheral corneal incision and corneal stromal pocket extended to the glass particle. This technique avoids creation of a corneal incision near the optical axis and prevents unwanted distortion of the corneal topography. The authors report this case to emphasize the importance of preserving corneal topography in corneal surgery.

Emerging antimicrobial resistance pattern of Helicobacter pylori in central Gujarat.

BACKGROUND: Antimicrobial resistance is a growing problem in H. pylori treatment. The study was intended to evaluate the prevalence of resistance amongst 80 H.pylori isolates cultured from biopsy taken during routine endoscopies in 2008-2011. MATERIALS AND METHODS: 855 gastro duodenal biopsies were collected and cultured on H.pylori selective medium (containing Brucella agar and Columbia agar (Hi media), with Skirrow's supplement (antibiotic supplement) and 7% human blood cells). H.pylori was isolated from 80 specimens. The antimicrobial susceptibility of H.pylori isolates was carried out by the Kirby Bauer technique against metronidazole (5 µg), clarithromycin (15 µg), ciprofloxacin (5 µg), amoxicillin (10 µg), tetracycline (30 µg), erythromycin (15 µg), levofloxacin (5 µg), and furazolidone (50 µg) (Sigma- Aldrich, MO). RESULTS: 83.8% isolates were resistant to metronidazole, 58.8% were resistant to Clarithromycin 72.5% were resistant to Amoxicillin, 50% to Ciprofloxacin and 53.8% to tetracycline. furazolidone, erythromycin and Levofloxacin showed only 13.8% resistance to H.pylori. Multi drug resistance with metronidazole+clarithromycin+tetracycline was 85%. For all the drugs Antimicrobial resistance rate was found higher in males compare to females. Metronidazole and amoxicillin resistance was found noteworthy in patients with duodenal ulcer (p=0.018), gastritis (P=0.00), and in reflux esophagitis (P=0.00). clarithromycin and tetracycline resistance was suggestively linked with duodenitis (P=0.018), while furazolidone, erythromycin and levofloxacin showed excellent sensitivity in patients with duodenitis (P value--0.018), gastritis (P=0.00) and reflux esophagitis (P=0.00). Resistance with metronidazole (P=0.481), clarithromycin (P=0.261), amoxicillin (P=0.276), tetracycline (P=0.356), ciprofloxacin (P=0.164) was not correlated well with Age-group and Gender of the patients. CONCLUSION: A very high percentage of patients were infected with metronidazole and clarithromycin resistant strains. The use of antibiotics for other indications seems to be the major risk factor for the development of primary resistance. High incidence should alarm the gastroenterologist while prescribing the eradication regimen.

Preparation, characterization and in vitro dissolution study of Nitrazepam: Cyclodextrin inclusion complex.

The objectives of this research were to prepare and characterize inclusion complexes of Nitrazepam with Hydroxypropyl-ß-cyclodextrin (HPßCD) and Sulfobutyl ether ß-cyclodextrin (SBEßCD) to study the effect of complexation on the dissolution rate of Nitrazepam, a water-insoluble drug. The phase solubility profile of Nitrazepam with Hydroxypropyl- ß-cyclodextrin and Sulfobutyl ether ß-cyclodextrin was an AP-type, indicating the formation of 2:1 stoichiometric inclusion complexes. Gibbs free energy values were all negative, indicating the spontaneous nature Nitrazepam solubilization and their value decreased with increase in the cyclodextrin concentration, demonstrating that the reaction conditions became more favorable as the concentration of cyclodextrins increased. Complexes of Nitrazepam were prepared with cyclodextrin using various methods such as physical mixing, kneading, spray-drying and lyophilization. The complexes were characterized by Differential scanning calorimetry, Fourier-transform infrared, scanning electron microscopy and powder X-ray diffraction studies. These studies indicated that a complex prepared by lyophilization had successful inclusion of the Nitrazepam molecule into the cyclodextrin cavity. Complexation resulted in a marked improvement in the solubility and wettability of Nitrazepam. Among all the samples, a complex prepared with Sulfobutyl ether ß-cyclodextrin by lyophilization had the greatest improvement in the in vitro rate of Nitrazepam dissolution. The mean dissolution time for Nitrazepam decreased significantly after preparing complexes. The similarity factor indicated a significant difference between the release profiles of Nitrazepam from complexes, physical mixtures and plain Nitrazepam. To conclude that, the tablets containing complexes prepared with Cyclodextrins had significant improvement in the release profile of Nitrazepam as compared to tablets containing Nitrazepam without cyclodextrin.