Blind esophageal brushing offers a safe and accurate method to monitor inflammation in children and young adults with eosinophilic esophagitis.

Patients with eosinophilic esophagitis (EoE) require frequent evaluation of mucosal inflammation via endoscopy. Instead of endoscopy, mucosal evaluation in adults with esophageal cancer and candidiasis is achieved using a cytology brush inserted through a nasogastric tube (NGT). We conducted a prospective cross-sectional study in children and young adults scheduled for routine esophagogastroduodenoscopy (EGD) where in Phase 1, we performed esophageal brushing through the endoscope under direct visualization and in Phase 2, we inserted the brush through a Cortrak® NGT prior to endoscopy. Eosinophil-derived neurotoxin (EDN) measured by ELISA in the samples extracted from brushes was validated as the sensitive biomarker. We collected 209 esophageal brushing samples from 94 patients and we found that EDN in brushing samples collected via EGD or NGT was significantly higher in patients having active EoE (n = 81, mean EDN 381 mcg/mL) compared with patients having gastroesophageal reflux disease (n = 31, mean EDN 1.9 mcg/mL, P = 0.003), EoE in remission (n = 47, mean EDN 3.7 mcg/mL, P = 0.003), or no disease (n = 50, mean EDN 1.1 mcg/mL, P = 0.003). EDN at a concentration of ≥10 mcg/mL of brushing sample was found to accurately detect active EoE. NGT brushing did not cause any significant adverse effects. We concluded that blind esophageal brushing using an NGT is a fast, less invasive, safe, and well-tolerated technique compared with EGD to detect and monitor EoE inflammation using EDN as the sensitive biomarker.

Does Ethnicity Influence Fractional Exhaled Nitric Oxide in Healthy Individuals?: A Systematic Review.

BACKGROUND: Fractional exhaled nitric oxide (Feno) is used clinically as a biomarker of eosinophilic airway inflammation. Awareness of the factors influencing Feno values is important for valid clinical interpretation. METHODS: We undertook a systematic review of PubMed, Cochrane Library, Scopus, and Web of Science databases and reference lists of included articles to evaluate whether ethnicity influences Feno values, and to determine if this influence affects clinical interpretation according to current guidelines. We included all studies that performed online Feno measurements on at least 25 healthy, non-Caucasian individuals, and examined the effect of ethnicity on Feno. RESULTS: From 62 potential studies, 12 studies were included. One study recruited only children (< 12 years of age), six studies recruited children and/or adolescents, four studies recruited adults only, and a single study involved children, adolescents, and adults. In total, 16 different ethnic populations representing 11 ethnicities were studied. Ethnicity was considered a significant influencing factor in 10 of the included studies. We found the geometric mean Feno to be above the normal healthy range in two studies. We also identified five studies in which at least 5% of participants had Feno results above the age-specific inflammatory ranges. CONCLUSIONS: Ethnicity influences Feno values, and for some ethnic groups this influence likely affects clinical interpretation according to current guidelines. There is a need to establish healthy Feno reference ranges for specific ethnic groups to improve clinical application.

Indigenous Respiratory Outreach Care: the first 18 months of a specialist respiratory outreach service to rural and remote Indigenous communities in Queensland, Australia.

OBJECTIVE: Respiratory diseases are a leading cause of morbidity and mortality in Indigenous Australians. However, there are limited approaches to specialist respiratory care in rural and remote communities that are culturally appropriate. A specialist Indigenous Respiratory Outreach Care (IROC) program, developed to address this gap, is described. METHODS: The aim of the present study was to implement, pilot and evaluate multidisciplinary specialist respiratory outreach medical teams in rural and remote Indigenous communities in Queensland, Australia. Sites were identified based on a perception of unmet need, burden of respiratory disease and/or capacity to use the clinical service and capacity building for support offered. RESULTS: IROC commenced in March 2011 and, to date, has been implemented in 13 communities servicing a population of approximately 43000 Indigenous people. Clinical service delivery has been possible through community engagement and capacity building initiatives directed by community protocols. CONCLUSION: IROC is a culturally sensitive and sustainable model for adult and paediatric specialist outreach respiratory services that may be transferrable to Indigenous communities across Queensland and Australia.

A randomized phase 3 trial of thalidomide and prednisone as maintenance therapy after ASCT in patients with MM with a quality-of-life assessment: the National Cancer Institute of Canada Clinicals Trials Group Myeloma 10 Trial.

We conducted a randomized, controlled trial comparing thalidomide-prednisone as maintenance therapy with observation in 332 patients who had undergone autologous stem cell transplantation with melphalan 200 mg/m2. The primary end point was overall survival (OS); secondary end points were myeloma-specific progression-free survival,progression-free survival, incidence of venous thromboembolism, and health-related quality of life (HRQoL). With a median follow-up of 4.1 years, no differences in OS between thalidomide-prednisone and observation were detected (respective 4-year estimates of 68% vs 60%, respectively; hazard ratio = 0.77; P = .18); thalidomide-prednisone was associated with superior myeloma-specific progression-free survival and progression-free survival (for both outcomes, the 4-year estimates were 32% vs 14%; hazard ratio = 0.56; P < .0001) and more frequent venous thromboembolism (7.3% vs none; P = .0004). Median survival after first disease recurrence was 27.7 months with thalidomide-prednisone and 34.1 months in the observation group. Nine second malignancies were observed with thalidomide-prednisone versus 6 in the observation group. Those allocated to thalidomide-prednisone reported worse HRQoL with respect to cognitive function, dyspnea, constipation, thirst, leg swelling, numbness, dry mouth, and balance problems. We conclude that maintenance therapy with thalidomide-prednisone after autologous stem cell transplantation improves the duration of disease control, but is associated with worsening of patient-reported HRQoL and no detectable OS benefit.

Lenalidomide plus melphalan without prednisone for previously untreated older patients with multiple myeloma: a phase II trial.

BACKGROUND: We conducted a phase II trial that evaluated the tolerability and efficacy of combining lenalidomide with melphalan in previously untreated patients with multiple myeloma who were not candidates for autologous stem cell transplantation. METHODS: After a run-in phase of 6 patients, we planned to conduct a randomized phase II selection-design trial that assessed 2 dose levels of lenalidomide, given days 1 to 21, combined with melphalan, given days 1 to 4, and every 28 days. Planned doses of melphalan were 9 mg/m(2)/d and respective doses of lenalidomide were 10 and 20 mg/d (M9L10 and M9L20). Coprimary endpoints were the frequency of dose-limiting Planned doses of melphalan were 9 mg/m(2)/d and respective doses of lenalidomide were 10 and 20 mg/d (M9L10 and M9L20). toxicities (DLT) and complete response (CR). RESULTS: Four patients received M9L10; all experienced DLTs, which resulted in closure of this cohort. When using the same schedule, we then sequentially tested M6L10 (melphalan 6 mg/m(2) on days 1 to 4 and lenalidomide 10 mg/d on days 1 to 21 every 28 days) (6 patients), M4L15 (melphalan 4 mg/m(2) on days 1 to 4 and lenalidomide 15 mg/d on days 1 to 21 every 28 days) (6 patients), and M5L10 (melphalan 5 mg/m(2) days 1 to 4 and lenalidomide 10 mg/d days 1 to 21 every 28 days) (34 patients). In each cohort, the DLT endpoint was reached because of severe and prolonged hematologic toxicity. At the final dose level, M5L10, 20 of 27 patients experienced DLTs within their first 3 cycles; among 10 patients who received at least 6 cycles, none achieved a CR. CONCLUSIONS: Combining lenalidomide plus melphalan without prednisone is associated with substantial hematologic toxicity that precludes cyclical administration of adequate drug doses.

RBx 7,796: A novel inhibitor of 5-lipoxygenase.

OBJECTIVE: To evaluate the pharmacological profile of RBx 7,796, a novel 5-lipoxygenase inhibitor. MATERIALS AND METHODS: RBx 7,796 was evaluated for 5- lipoxygenase inhibitory potential using human recombinant enzyme and profiled for selectivity against 12 and 15 lipoxygenase. RBx 7,796 was evaluated in cell based assay for inhibition of A23,187 induced LTB(4) release from isolated neutrophils. Ex vivo activity was evaluated for inhibition of A23,187 induced LTB(4) release in blood from treated rats. In vivo efficacy of RBx 7,796 was profiled in LPS induced neutrophilia model in rats and also in ovalbumin induced bronchoconstriction and airway inflammation models in guinea pigs. RESULTS: RBx 7,796, a novel chemotype, showed competitive inhibition of 5-lipoxygenase enzyme with an IC(50) of 3.5 +/- 1.1 microM. RBx 7,796 offered >100 fold selectivity against other related enzymes - 12 and 15 lipoxygenase. RBx 7,796 inhibited release of LTB(4) from human and rat neutrophils in vitro. Upon administration to rats, RBx 7,796 inhibited A23,187 induced LTB(4) release from rat neutrophils. Upon repeated administration, dosed once daily, RBx 7,796 inhibited LPS induced neutrophil influx in rat airway. RBx 7,796 also inhibited allergen induced bronchoconstriction and eosinophil influx in guinea pig airway in a dose dependent manner. CONCLUSION: The results suggest that RBx 7,796, a novel chemotype, is an orally efficacious inhibitor of 5-lipoxygenase enzyme that is effective against both neutrophilic and eosinophilic airway inflammation and shows potent inhibition with once daily administration.

Reduction of hematotoxicity and augmentation of antitumor efficacy of cyclophosphamide by dopamine.

Modulatory effects of dopamine (DA) on hematotoxicity and antitumor efficacy of cyclophosphamide (CY) were studied in Swiss mice bearing transplantable Ehrlich ascites carcinoma (EAC). DA was administered i.p. at a dose of 50 mg/kg/day for 5 consecutive days beginning day 3 after tumor transplantation. CY (200 mg/kg i.p.) was injected 24 hour after completion of DA treatment. DA pretreatment reduced the suppressive effects of CY on hemoglobin, RBC, total WBC, neutrophil, platelet, and bone marrow nucleated cell counts. Likewise, DA partially prevented the CY-induced fall in pluripotent (CFU-S12) and lineage-specific stem cells for granulocytes (CFU-C) in bone marrow. Moreover, mice receiving a combination of DA and CY illustrated greater reduction in tumor volume, viable tumor cell count and mitotic index along with upregulation of tumor cell apoptosis than CY-only group. As a result, the former group demonstrated prolonged hosts survival. Thus, DA protected to a great extent the hematopoietic cells of tumor bearing hosts from the suppressive action of CY and concomitantly augmented its antitumor efficacy resulting in improved hosts survival.

Decrease in platelet serotonin level in uterine cervix cancer patients.

Platelet serotonin (5-HT) concentration was measured by HPLC with electrochemical detection in 46 women suffering from cancer of the uterine cervix and 16 matched controls. About 53% reduction (p < 0.05) was recorded in platelet 5-HT level in cancer patients against a control value of 1.29 +/- 0.16 (mean +/- S.E.) nmol per 10(9) platelets. Depletion of intraplatelet 5-HT was positively correlated with clinical stage of the disease although a modest rise (p > 0.05) in platelet 5-HT was observed in patients at stage I. Serotonin release from platelets following activation with thrombin was considerably increased in cancer patients (38.2% compared to 17.4% in controls). The results demonstrate progressive depletion of intraplatelet 5-HT in cervical cancer patients. In addition, their platelets release more 5-HT than the controls upon activation by thrombin.

A soxA gene, encoding a diheme cytochrome c, and a sox locus, essential for sulfur oxidation in a new sulfur lithotrophic bacterium.

A mobilizable suicide vector, pSUP5011, was used to introduce Tn5-mob in a new facultative sulfur lithotrophic bacterium, KCT001, to generate mutants defective in sulfur oxidation (Sox(-)). The Sox(-) mutants were unable to oxidize thiosulfate while grown mixotrophically in the presence of thiosulfate and succinate. The mutants were also impaired in oxidizing other reduced sulfur compounds and elemental sulfur as evident from the study of substrate oxidation by the whole cells. Sulfite oxidase activity was significantly diminished in the cell extracts of all the mutants. A soxA gene was identified from the transposon-adjacent genomic DNA of a Sox(-) mutant strain. The sequence analysis revealed that the soxA open reading frame (ORF) is preceded by a potential ribosome binding site and promoter region with -10- and -35-like sequences. The deduced nucleotide sequence of the soxA gene was predicted to code for a protein of 286 amino acids. It had a signal peptide of 26 N-terminal amino acids. The amino acid sequence showed similarity with a putative gene product of Aquifex aeolicus, soluble cytochrome c(551) of Chlorobium limicola, and the available partial SoxA sequence of Paracoccus denitrificans. The soxA-encoded product seems to be a diheme cytochrome c for KCT001 and A. aeolicus, but the amino acid sequence of C. limicola cytochrome c(551) revealed a single heme-binding region. Another transposon insertion mutation was mapped within the soxA ORF. Four other independent transposon insertion mutations were mapped in the 4.4-kb soxA contiguous genomic DNA region. The results thus suggest that a sox locus of KCT001, essential for sulfur oxidation, was affected by all these six independent insertion mutations.

Cloning and characterization of a high-copy-number novel insertion sequence from chemolithotrophic Thiobacillus ferrooxidans.

Two distinct families of repetitive DNA elements (1.4 and 1.2 kb) were identified from S1 nuclease-treated genomic DNA of four strains of Thiobacillus ferrooxidans. The 1.4-kb fragment hybridized with IST2, an insertion sequence of T. ferrooxidans. The 1.2-kb fragment was cloned and sequenced. The sequence (IST445), 1219 bp in length, with features characteristic of an insertion element, has a terminal inverted repeat of 8 bp, which can be further extended to 23 or 48 bp with 9 and 26 mismatches, respectively. It displays 54.4% identity in 967 nucleotides of overlap with ISAE1 of Alcaligenes eutrophus. The IST445 contains three open reading frames which have codon usage almost similar to 56 different coding genes of T. ferrooxidans. In Southern blots of restricted genomic DNAs probed with IST445, each of the several strains of T. ferrooxidans gives a distinctive fingerprint. IST445 is present in the range of 10-20 copies per genome in the four strains studied.

Mitomycin C induced inhibition of adrenal steroidogenesis in female rats.

Mitomycin C (MC), an antibiotic which depresses DNA synthesis causes suppression of enzyme delta 5 3 beta-hydroxysteroid dehydrogenase (delta 5 3 beta OHD) and glucose-6 phosphate dehydrogenase (G-6 PD) in the rat adrenal tissue. The treatment resulted in a fall in DNA content together with an accumulation of cholesterol and ascorbic acid in the gland. The results suggest a diminution in adrenal steroid biogenesis similar to gonadal inhibition previously reported.

Quinalphos-induced suppression of spermatogenesis, plasma gonadotrophins, testicular testosterone production, and secretion in adult rats.

Quinalphos (O,O-diethyl-O-[quinoxalinyl-(2)-thionophosphate]) is a well-known organophosphorus insecticide used extensively in agriculture that adversely interferes with the activity of testicular steroidogenic enzymes in rats. To investigate its effects on spermatogenesis, the other function of testes, quantitative evaluation of different varieties of germ cells at stage VII of the seminiferous epithelium cycle, namely, type A spermatogonia (ASg), preleptotene spermatocytes (pLSc), midpachytene spermatcytes (mPSc), and step 7 spermatids (7Sd), along with the radioimmunoassay of plasma FSH, LH, testosterone, and testicular testosterone, were performed in Wistar rats following treatment with quinalphos (250 micrograms/kg, ip) for approximately one (13 days) and two cycles (26 days) of the seminiferous epithilium. Massive degeneration of all varieties of germ cells at stage VII, remarkable reduction in the sperm count, and significant reductions in plasma concentrations of FSH and testosterone, along with testicular testosterone, were observed after quinalphos treatment. Significant reduction in the plasma concentration of LH was observed only after treatment for two cycles. Administration of human chorionic gonadotrophin for 26 days in rats injected with quinalphos partially prevented the degeneration of germ cells and increased testosterone production. It is suggested that quinalphos may have a suppressive influence on gonadotrophin release but its direct detrimental action at the level of the testes may also be responsible for the observed changes in spermatogenesis and in testicular testosterone production in rats.

Suppressive effect of quinalphos on the activity of accessory sex glands and plasma concentrations of gonadotrophins and testosterone in rats.

Biochemical estimation of prostatic acid phosphatase and fructose content in accessory sex glands, along with radioimmunoassay of plasma gonadotrophins (FSH and LH) and testosterone were performed in Wistar rats following treatment with quinalphos, an organophosphorus insecticide, for 13 and 26 days. Prostatic acid phosphatase activity and fructose content of the accessory sex glands, and plasma levels of testosterone and FSH were significantly lower in all rats treated with quinalphos. However, the degree of inhibition was more extensive in the 26 day-treatment group who, in addition also exhibited a significant reduction in relative weights of the testes and accessory sex organs, and plasma levels of LH. All these adverse effects of quinalphos were prevented when exogenous HCG was administered in concomitant with the insecticide for 26 days. These results suggest that quinalphos may exert a suppressive effect on the functional activity of accessory sex glands by decreasing testicular testosterone production following inhibition of pituitary gonadotrophins release.

Antifertility effect of Piper betle Linn. extract on ovary and testis of albino rats.

Chronic administration (sc) of the extract of the stalk of P. betle at 30 mg/kg body weight daily for 21 days produced significant decrease in oestrogen and androgen dependent target organ weights along with increase in cholesterol in adrenal, ovary and testis. Acid and alkaline phosphatase activities in serum, liver and kidney did not exhibit any toxic effect. There was marked change in morphology of testis and ovary. Vaginal smear showed prolonged dioestrus in treated female. The treated male showed decreased number and motility of sperm. Both male and female remained infertile after treatment suggesting antifertility activity of the extract on both sexes of albino rats.

Effect of aldrin on accessory sex glands and plasma testosterone levels in rats.

The effects of aldrin, an organochlorine insecticide, on accessory sex glands and plasma testosterone levels in rats were studied. The aldrin was administered i.p. for 13 days and 26 days at a dose of 150 micrograms/kg. Relative weights of prostate, seminal vesicles and coagulating glands were significantly decreased in the treated rats compared to those in controls. In addition, there was a significant fall in acid phosphatase activity in prostate and fructose content in accessory sex glands was also observed in treated animals. Plasma testosterone values showed a decrease with the duration of treatment. HCG supplementation with aldrin treatment prevented all those untoward effects of aldrin in experimental rats.

Effect of aldrin on spermatogenesis, plasma gonadotrophins and testosterone, and testicular testosterone in the rat.

Quantitative evaluation of the different varieties of germ cells at stage VII of the seminiferous epithelium cycle, namely type-A spermatogonia (ASg), preleptotene spermatocytes (pLSc), mid-pachytene spermatocytes (mPSc) and step 7 spermatids (7Sd), along with radioimmunoassay of plasma gonadotrophins (FSH and LH), testosterone and testicular testosterone were performed in Wistar rats following treatment with aldrin (polycyclic chlorinated hydrocarbon insecticide) for approximately one (13 days) or two cycles (26 days) of the seminiferous epithelium. Extensive degeneration of all varieties of germ cells at stage VII, reduction in the sperm count and significant reductions in plasma concentrations of LH and testosterone were observed following aldrin treatment. The reduction in plasma concentrations of FSH was statistically significant only after treatment for two cycles. The inhibitory effect of aldrin on plasma gonadotrophins, testosterone levels, testicular testosterone content and numbers of 7Sd and ASg was maximum after treatment for two cycles. Administration of human chorionic gonadotrophin along with aldrin treatment for two cycles partially prevented the degeneration of germ cells and enhanced testosterone production. The results indicate that aldrin may have a direct inhibitory influence on gonadotrophin release, but the possibility of a direct action of the insecticide at the level of the testes is also discussed.

Effect of quinalphos on testicular steroidogenesis in rats.

The effect of quinalphos (250 micrograms/kg i.p.) an organophosphorus insecticide treatment for 13 and 26-days on the testicular steroidogenic enzymes viz. 3 beta-Hydroxysteroid Dehydrogenase and 17 beta-Hydroxysteroid Dehydrogenase, as well as cholesterol content and histology of the testes of the Wistar strain rats was studied. The time duration of 13 days is approximately equivalent to one cycle of the seminiferous epithelium in Wistar strain rats. Treatment of quinalphos for 13 days failed to produce any effect on the relative weights of the testes and accessory sex glands. However, significant inhibition of 3 beta-HSD activity and increased cholesterol level in testis were observed. The rats treated for 26 days similarly showed a highly significant inhibition of the activity of both 3 beta-HSD and 17 beta-HSD. The relative weights of the testes and accessory sex glands were also significantly reduced. Histological examination of the testis revealed that quinalphos treatment produced detrimental changes in the seminiferous epithelium. Treatment with quinalphos for 13-days produced no toxic effect with the exception of a significant increase in serum alkaline phosphatase. However, after 26-days of treatment toxicity was significantly increased as reflected on serum transaminases, phosphatases and blood urea levels of rat. Present study indicated that quinalphos impairs testicular functions in rats.

Suppression of testicular steroidogenesis in rats by the organochlorine insecticide Aldrin.

Treatment with Aldrin, an organochlorine insecticide, for 13 and 26 days caused suppression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activities, along with accumulation of cholesterol in the testicular tissues of adult rats. The same treatment also resulted in a reduction in the nuclear diameter of Leydig cells (LCND) and diameter of seminiferous tubules. A decrease in the weight of testes, seminal vesicles and ventral prostate was also noted. HCG administration in the long-term (26 days) treatment restored the steroidogenic enzymes activity and the nuclear diameter of the Leydig cells. It also reduced the accumulation of tissue cholesterol towards the vehicle-injected controls. The inhibition of steroidogenesis in the testes possibly reflects a decrease in pituitary gonadotrophin release after the treatment with Aldrin.