Pseudoplatystoma are very important species due to both their high commercial value and their ecological role as voracious predators. They undertake lengthy migratory movements during their life-cycle, this including reproductive migration which occurs from October to December in the rainy season. In the present study, seven microsatellite loci were analyzed to access genetic variability in two samples of P. reticulatum from the Upper Paraguay Basin. The loci were highly polymorphic (mean = 7.28). According to all analysis, the two samples of P. reticulatum revealed pronounced genetic differentiation. Fst value was 0.2290, Rst value 0.1067 and AMOVA 22.90% (Fst ) and 10.67% (Rst ), all being highly significant (p < 0.001). The division of the fishes into two groups was confirmed by microsatellite multi-locus Bayesian assignment testing. The results obtained present evidence of genetic structuring in a P. reticulatum population. Catfishes of the genus  Pseudoplatystoma belong to the family Pimelodidae, the 93 species of which occurring in the major river basins of South America . Studies using microsatellites have revealed pronounced genetic differences even among populations isolated by short geographic distances (Pseudoplatystoma reticulatum (“cachara”) collected in the Upper Paraguay River Basin, by using seven microsatellite loci originally developed for P. corruscans to access the genetic diversity of these samples. These data can be used as subsidies for setting up management policies for the development and conservation of these species.Microsatellite markers have been extensively used in studies on the genetics of fish populations. These markers have a co-dominant inheritance pattern, a high degree of polymorphism, and allow for easy analysis through experiments involving the polymerase chain reaction (PCR) (P. reticulatum were caught by gill nets at two points on the Paraguay River Basin. 31 specimens were collected from the Paraguay River (PRP) (16° 04' 00" S 57° 41' 00" W) and 21 from the Jauru River (PRJ) (15° 51' 00" S 58° 27' 00" W). Fin clips were the source of nuclear DNA. These were collected from freshly caught fish and immediately preserved in 95% ethanol. After collection, the fishes were sold. For the extraction of genomic DNA, about 0.1 mg of tissue was incubated in 200 μL of 5% Chelex (Sigma®) at 65 °C overnight.A total of 52 adult specimens of P. reticulatum samples were screened for variation at each of the seven microsatellite loci. Five of these loci have already been described by 2, 0.2 U of Taq-Pht DNA polymerase, 1X PCR buffer  and water. We used the following PCR profile for the loci Pcor01, Pcor02, Pcor05, Pcor08, Pcor21, Pcor23 and Pcor28: initial denaturation at 95 °C for 5 min, 30 cycles of 10 s at 95 °C, 15 s at an annealing temperature of 55 °C, 15 s at 72 °C and a final extension at 72 °C for 10 min. For the locus Pcor10, the PCR profile consisted of an initial denaturation at 95 °C for 5 min, 30 cycles of 30 s at 95 °C, 30 s at an annealing temperature of 48 °C, 30 s at 72 °C and a final extension at 72 °C for 10 min. Amplified products were resolved on 6% polyacrylamide gels stained with silver nitrate. Microsatellite alleles were identified by their size in base-pairs. Allele lengths were estimated by comparison with a 10 bp ladder (10 pb DNA Ladder - Invitrogen), using Kodak Digital Science 1D software.The HE, HO), inbreeding coefficient (FIS) and gene flow (Nm = 0.25(1-FST)/FST) were obtained with POPGEN 1.32 software , and their significance was tested with 1,000 permutations. Corrections of the significance level for multiple tests were performed following the Bonferroni procedure  was estimated by using the ‘no admixture' ancestral model with correlated alleles, thereby allowing for maximal population resolution, with K ranging from 1 to 6. Three independent runs of 500,000 Markov chain Monte Carlo (MCMC) generations and 100,000 generations of ‘burn-in' were used for each value of K. The true number of populations is expected to be the value of K that maximizes the estimated model log-likelihood, log (P(X|K))  to 11 (Pcor08), and allelic richness from 1.722 (Pcor28) to 9.503 (Pcor08) (Pcor28) to 0.870 (Pcor01). A total of 51 alleles were detected, of which 26 were private (Pcor28 - PRJ) to 0.8615 (Pcor01 - PRJ) and from 0.0556 (Pcor28 - PRJ) to 0.5556 (Pcor08 - PRJ and PRP), respectively  . Nei gen private . The pri private . Expecteectively . Signifiectively . The occanalyzed .P. reticulatum, estimated through the FST index for all the loci, was 0.2290 and was statistically highly significant , thereby showing the existence of strong genetic differentiation among the analyzed samples. The RST index estimated for all the loci was 0.1067, which was also statistically highly significant .Genetic differentiation between the two populations of Individual multi-locus genotypes were used to assign individuals to their respective population of origin. On considering the correct assignment of all individuals, 98.3% were correctly assigned to the PRP population and 99.1% to the PRJ population.FST = 77.10% and RST = 89.33%. The values for variability between populations were FST = 22.90% and RST = 10.67%. These were highly significant (p < 0.0001), thus revealing the strong structure of P. reticulatum populations.Hierarchical AMOVA revealed that most total genetic variance was to be found within populations K = 2 populations maximizing the estimated log likelihood in the model , consistent with the mean number observed in other fish species . HoweverA significant deviation of HWE (p < 0.05) was observed in seven loci within two samples analyzed . HWE depFIS was calculated for all loci. The results acquired displayed significant departure from HWE  and moderate (0.1067) values, respectively (p < 0.025), thus suggesting there is a strong genetic structure. The differences of values between indices were probably due to differences in the mutation models on which they were based. While FST is based on IAM, RST is based on SMM. The RST index may be the best for microsatellite analysis, and it is expected that RST values under a strict SMM pattern would be higher than those of FST 9GC(TC)9-; Pcor05 -(TC)8CC(TC)15- and Pcor10 -(GTCG)15(GT)9(CC-).There was significant populational structuring between the two samples of e of FST . HoweverMM model . This prP. reticulatum on using both indices (FST and RST), thereby depicting the occurrence of strong genetic structuring. The values of molecular variation among populations were 22.90% (FST) and 10.67% (RST), both with highly significant p values (p < 0.0001). Through structure analysis without admixture, it was shown that both populations were genetically distinct, with K = 2 populations maximizing the estimated log-likelihood in the model , 98.3% and 99.1% of the individuals were correctly assigned to the location from which they were sampled . These results are consistent with the values found by P. corruscans populations . FST ~ 0.1).The genetic flow of all loci in the two populations was estimated as ulations . Thus, oP. reticulatum analyzed. In five of these, frequency was higher than 10% (P. reticulatum analyzed. Furthermore, all the tests employed confirmed the strong structure in these populations. Thus, our work renders preliminary evidence of the genetic structure in P. reticulatum. As the results presented are consistent with those obtained for P. corruscans (P. reticulatum also presents homing behavior.Twenty-six private alleles were found in the two populations of than 10% . The exiP. reticulatum, the present data constitute a particularly important element when contemplating their management and conservation. It also places in evidence, the importance of preserving each population to further a positive outcome in the genetic conservation of these species.The ability to identify and define biological populations is crucial for taking informed decisions concerning conservation and management . Conside


Oropharyngeal squamous cell carcinoma (OPSCC) is associated with the sexually transmitted human papillomavirus (HPV), smoking and alcohol. In Greenland, a high rate of HPV-induced cervical cancer and venereal diseases are found, which exposes the population for high risk of HPV infection. In Greenland, only girls are included in the mandatory HPV vaccination program.To investigate the annual incidence of OPSCC and the proportion of HPV-associated OPSCC (HPV+ OPSCC) in Greenland in 1994–2010.At Rigshospitalet, University of Copenhagen, we identified all Greenlandic patients diagnosed and treated for OPSCC from 1994 to 2010. Sections were cut from the patient's paraffin-embedded tissue blocks and investigated for p16 expression by immunohistochemistry. HPV analyses were performed with 2 sets of general HPV primers and 1 set of HPV16-specific primer. HPV+ OPSCC was defined as both >75% p16+ cells and PCR positive for HPV.Of 26 Greenlandic patients diagnosed with OPSCC, 17 were males and 9 were females. The proportion of HPV+ OPSCC in the total study period was 22%, without significant changes in the population in Greenland. We found an increase in the proportion of HPV+ OPSCC from 14% in 1994–2001 to 25% in 2002–2010 (p=0.51). Among males from 20 to 27% (p=0.63) and in females from 0 to 20% (p=0.71). The annual OPSCC incidence increased from 2.3/100,000 (CI=1.2–4.2) in 1994–2001 to 3.8/100,000 (CI=2.4–6.2) in 2002–2010: among males from 2.4/100,000 (CI=1.0–5.7) to 5.0/100,000 (CI=2.9–8.9).Even though the population is at high risk of HPV infection, the proportion of 22% HPV+ OPSCC in the total study period is low compared to Europe and the United States. This might be explained by our small study size and/or by ethnic, geographical, sexual and cultural differences. Continuing observations of the OPSCC incidence and the proportion of HPV+ OPSCC in Greenland are needed. Head and neck cancer is the fourth most common malignant cancer worldwide and is known to be associated with high consumption of alcohol, tobacco and human papillomavirus infection (HPV) , 2. InciIn Greenland, the annual incidence rate of OPSCC from 1994 to 2003 was 2.1/100,000 accounting for 11% of all head and neck cancers in Greenland . The devHPV is known to cause cervical squamous cell carcinoma (CSCC). A recent study finds a significant similarity in the miRNA profile of HPV+ CSCC and HPV+ OPSCC . Other rThe carcinogenic effect of HPV is caused by the expressed oncoproteins E6 and E7 that inactivate the tumour suppressor genes p53 and retinoblastoma protein (pRB) leading to uncontrolled growth and a reciprocal up-regulation of the tumour suppressor protein p16 . p16 is In Greenland, the HPV vaccine is currently administered, free of charge, only to girls aged 12–27 years as a part of the Danish and Greenlandic free mandatory national vaccination programme . IntroduThe aims of the present study are to investigate the proportion of HPV+ OPSCC and OPSCC incidence in the Greenlandic population and to find potential changes in the time period 1994–2001 compared to 2002–2010. The perspectives of HPV vaccination of boys will also be presented.This study is retrospective, including patients born and living in Greenland who were biopsied and diagnosed with OPSCC  in the period 1994–2010. The patients were biopsied in Greenland or at Rigshospitalet, and all were transferred for further treatment at Rigshospitalet, which serves as a tertiary referral hospital for Greenland. The patients and their medical records were identified by the patient registry at Rigshospitalet and the diagnosis verified by the Danish Pathology Registry. Information regarding tobacco and alcohol consumption, and the stage of cancer was obtained from medical records and from the Danish Head and Neck Cancer Group database (DAHANCA) . The patFormalin-fixed paraffin-embedded (FFPE) tumour tissue blocks  of OPSCC from the Greenlandic patients were collected at the Department of Pathology, Rigshospitalet, where all histological specimens from Greenland are examined. Tumour sections of 4 µm were cut, stained with HE and the OPSCC diagnosis was verified by a specialized head and neck pathologist. For p16 immunohistochemistry, the representative slides were incubated with p16 antibody  and stained on a BenchMark ULTRA  using the UltraView detection system . The p16Four sections of 10 µm were cut from the FFPE OPSCC specimens and were transferred to Eppendorf tubes. The genomic DNA was extracted by the QIAamp procedure  after prDNA quality of the extracted DNA samples was controlled by the expression of the housekeeping gene glyceraldehyde 3-phosptate dehydrogenase (GAPDH) . Only DNWe defined HPV-associated (HPV+) OPSCC by 2 criteria: The p16 immunohistochemistry should be of score 4+ (>75% positive cells) and the specimen should be PCR positive with one of the HPV-specific primer sets for OPSCC . As smokStatistical analyses were performed using SPSS v. 20 from IBM. Fisher's exact test was used for testing proportions. Significant results were defined as a p-value of ≤0.05. Annual incidence rates were age-standardized using the world population and calculated using the mean population of Greenlandic inhabitants as the denominator in 2 study time intervals: 1994–2001  and 2002–2010  . RothmanThe study followed the Helsinki II declaration and was approved by the Commission for Scientific Research in Greenland. The Danish Data Protection Agency approved the use of the patient's data.A total of 30 patients diagnosed with OPSCC in Greenland from 1994 to 2010 were identified. Of these, 4 patients were not born in Greenland, leaving 26 ethnical Greenlanders with available OPSCC specimens for the study. In the total study period 1994–2010, we found 11/26 (42%) of the OPSCC specimens to be p16+ , while 3/26 (12%) were of score 1+ and 2+ (<50% staining). The rest of the OPSCC specimens presented no p16 staining. The median age at diagnosis of the patients with HPV+ OPSCC was 47 years compared to the age of 63 years when diagnosed with HPV− OPSCC (p=0.3) .In 1994–2001, the annual OPSCC incidence was 2.3/100,000 (CI=1.2–4.2), in males 2.4/100,000 (CI=1.0–5.7) and in females 2.2/100,000 (CI=0.8–5.8) . The annIn 1994–2001, the proportion of p16+ OPSCC was 22%  and the proportion of HPV+ OPSCC was 14% . In 2002–2010, the proportion of p16+ increased to 53% (p=0.14), in males to 58% (p=0.56) and in females to 40% (p=0.12), while the proportion of HPV+ OPSCC increased to 25% (p=0.51), in males to 27% (p=0.63), and in females to 20% (p=0.71).Data on tobacco and alcohol consumption were available for 21 of the 26 OPSCC patients. Of the HPV+ OPSCC patients, 2/4 (50%) were smokers and 1/4 (25%) consumed more than 7 units of alcohol per week, while among the HPV− OPSCC patients, 15/17 (88%) were smokers (p=0.15) and 12/17 (71%) consumed more than 7 units per week (p=0.13) .In our study, we found an increase in the proportion of HPV+ OPSCC from 14% in 1994–2001 to 25% in 2002–2010, and an overall HPV+ OPSCC proportion of 22%. Compared to Europe, the proportion is low. A study conducted in the United Kingdom using similar methods and definitions of HPV+ examined 108 OPSCC patients and found an increase in the proportion of HPV+ OPSCC from 15% in 1988 to 57% in 2009 . InternaOur lower proportion of 22% HPV+ OPSCC, compared to western countries, may be explained by the small study group consisting of only 26 Greenlandic patients. Also high smoking and alcohol consumption was so dominant in Greenland that the effect of the HPV maybe is difficult to observe . In 1993It is known that HPV is sexually transmitted and the rate of oral infection with HPV16 correlates with the number of sexual partners, oral sex and open-mouthed kissing. Also, from a register study we know that partners to women with positive pap-smears for CSCC are at higher risk of OPSCC , 33. TheThe proportion of 22% HPV+ OPSCC cases demonstrates that HPV infection is more common in the Greenlandic patients with OPSCC compared to healthy people. American studies conducted in 2009–2010 among 3,977 healthy Americans found 1.3% to have oral HPV infection , and anoWe find a discrepancy between the proportion of 42% p16+ OPSCC cases and only 22% PCR-positive OPSCC cases. These p16+ and PCR-negative OPSCC cases may have been induced by other factors such as other viral infections or mutations  that might inactivate pRB and thereby lead to an up-regulation of p16 , 38. p16Males accounted for two-thirds of the OPSCC cases. The alcohol and tobacco consumption were lower in patients with HPV+ OPSCC compared to HPV− OPSCC patients, and the HPV+ OPSCC cases were diagnosed 16 years earlier (median 47 years) compared to HPV− OPSCC (median 63 years). These results are in accordance with international findings , 43, 44.Possible treatments of HPV+ OPSCC today include surgery and radiotherapy with or without concomitant chemotherapy. But it is possible that vaccines administered today will prevent HPV+ OPSCC in the future. The quadrivalent vaccine covers the oncogenic HPV16 and HPV18 viruses and the non-oncogenic, papilloma inducing HPV6 and HPV11 viruses, and the bivalent vaccine covers HPV16 and HPV18 viruses. In a multi-ethnic, multinational , 45 4-yeFrom 1988 to 2004, the annual incidence of HPV+ OPSCC in the United States has increased 225%, and it is especially found in males aged 30–50. The incidence of HPV+ OPSCC is predicted to surpass the incidence of HPV+ CSCC in the year 2020 . As a reWe found an increase in the annual OPSCC incidence in Greenland from 2.3/100,000 (CI=1.2–4.2) in 1994–2000 to 3.8/100,000 (CI=2.4–6.2) in 2001–2010; among males from 2.4/100,000 (CI=1.0–5.7) to 5.0/100,000 (CI=2.9–8.9). We found an increase in the proportion of HPV+ OPSCC from 14 to 25% in the same interval (p=0.51). Patients suffering from HPV+ OPSCC were diagnosed at an earlier age (47 years compared to 63 years), and there was a trend towards lower consumption of alcohol (p=0.13) and tobacco (p=0.15) compared to patients with HPV− OPSCC. The overall proportion of 22% HPV+ OPSCC was lower compared to Europe and the United States, possibly due to small sample sizes and/or geographical, sexual, ethnic or cultural differences. The Greenlandic population is still at high risk of HPV infection as demonstrated by the high incidence of HPV-induced CSCC. Continuing studies of the OPSCC incidence and the proportion of HPV+ OPSCC in Greenland are needed.


Objective: The aim of this study was to investigate thyroid function tests in Gestational Diabetes Mellitus (GDM) and pre-gestational DM and control group.Methodology: There were 61 pregnant diabetic women in study group and 35 pregnant women in control group. Serum T4, T3, T3RU, FTI, TSH and Anti TPO Ab were assessed in each person.Results: About 36% of patients had GDM and 64% pre-gestational DM. Thyroid dysfunction was detected in 18% of study group compared with 8.6% of control group (P = 0.2). There was Thyroid dysfunction in 4.5% of GDM and 25.6% of pregestational DM (P = 0.045). There was no statistically significant difference between thyroid dysfunction in GDM group and control group (P=0.99).27% of GDM and 36% of pregestational DM and 23% of control group had positive titer of Anti TPO Ab without statistically significant differences among the three groups.Conclusion: Thyroid dysfunction is prevalent in women with pre-gestational DM so, thyroid function should be evaluated in these patients during pregnancy. Rate of thyroid dysfunction in GDM patients is similar to normal pregnant control women. High prevalence of positive titer of TPO Ab was seen in diabetic and non-diabetic pregnant women. It is classified as gestational diabetes (diabetes which was first diagnosed during pregnancy) or pre-gestational diabetes. GDM occurs in 2-6% of pregnant women in North America that is estimated 10 times more than pre-gestational diabetes.3-5About 10-15% of pregnant women have thyroid dysfunction during the first half of pregnancy, which may be hypothyroidism or hyperthyroidism.6 However, the prevalence of thyroid dysfunction in pregnant women with type I diabetes is about three times more than general population and subclinical hypothyroidism is more prevalent.7In Iran, the prevalence of gestational diabetes has been reported between 4.7-7.4% in different regions that according to the new ADA criteria, the prevalence of gestational diabetes will be much higher than the above numbers.8 Some studies have reported high incidence of hypothyroxinemia and high Anti TPO titer in GDM.9 Prevalence of high titer of Anti TPO Ab in healthy pregnant women and type 1 diabetes also have significant difference.10 Delayed diagnosis of hypothyroidism in pregnancy leads to congenital malformations and respiratory distress in newborns of these mothers.11In some studies 40% of pregnant women with type I diabetes may also have thyroid dysfunction simultaneously.12 When anti-thyroid antibodies are positive, there is risk of premature rupture of amniotic sac and preterm labor.13 Some studies have showed that even without the overt thyroid dysfunction, autoimmune thyroid disease increases abortion rates up to 3 to 5 times.14Furthermore mother's diabetes during pregnancy can influence the secretion of T3 or conversion of T4 to active T3 in the fetus and premature newborns.The purpose of this study was to evaluate thyroid function tests and thyroid autoantibody level in pregnant patients with diabetes  and comparison with a non-diabetic control group in Ahvaz (Western South of Iran).This is a cross sectional prospective study. The study population was pregnant women with diabetes  who were referred to Ahvaz Golestan Hospital Endocrinology Clinic from 2004 to 2006. Diabetic patients were aged 19 to 47 years. Thirty five non-diabetic pregnant women were also considered as control group. Sequential sampling was conducted. Written consent was obtained from all patients and the study was approved by the Ethics Committee of Ahvaz Jundishapur University of Medical Science.In this study T3, T4, TSH, T3RU and Anti TPO Ab were measured in all GDM and pre-gestational diabetic pregnant women and control group. T3, T4, T3rU, FTI hormones was measured by RIA method, TSH by IRMA, TPO Ab by ELISA, Counter. Kits used in this laboratory was Immunotech product of A Beckman coulter company made in America.Pre-gestational diabetes had been distinguished before pregnancy. GDM in pregnant women had been considered if two of the four criteria were detected following 100 g glucose tolerance test:.In the present study individuals with normal levels of T4 and T3 and TSH <0.3mlU / L had been considered as sub clinical hyperthyroidism and individuals with normal T4 and T3 and TSH> 4mlU / L as sub clinical hypothyroidism. High T4 and TSH <0.3 as hyperthyroidism and T4 <4.8 and TSH>4 as hypothyroidism.Questionnaire included type of diabetes , thyroid disease history, history of drug consumption for thyroid dysfunction, gestational age at diagnosis was completed for each patient. The data obtained in this study were analyzed by statistical software SPSS15.This study was performed on 61 diabetic pregnant patients. Among them 22 patients (36%) had GDM and 39 (64%) had pre-gestational diabetes (20% type 1 and 80% type 2 diabetes). Control group were 35 non-diabetic pregnant women. Diabetic patients were aged 19 to 47 years. The overall mean age was 30±6.5 years in all diabetic patients and 28.9 ± 5.53 years in GDM and 30.6±7.10 years in pre-gestational diabetes. Mean age of control group was 27.7±4.25. Among 22 patients with GDM, 21 patients (95.5%) had normal thyroid function and one patient (4.5%) had subclinical hypothyroidism. Among 39 patients with pre-gestational diabetes, 29 cases (74.4%) had normal thyroid function and 10 cases (25.6%) had thyroid dysfunction. In control group among 35 patients, three patients (8.6%) had thyroid dysfunction.Overall Subclinical hypothyroidism was the more prevalent thyroid dysfunction in the three groups. Thyroid dysfunction was significantly higher in pre-gestational than GDM group (P = 0.037). Thyroid dysfunction in GDM and pre-gestational group did not have significant difference with control group . Previous history of thyroid disease were found in 16 cases (16.3%) and all of them were in pre-gestational group.Types of thyroid dysfunction in diabetic patients and control groups are shown in High titer of TPO Ab was detected in 27% of the GDM group, 36% of pre-gestational DM and 23% of control group. There was no significant differences in Anti TPO positive titer between three groups (p=0.188).6 Patterns of ontogeny of fetal cerebral cortex deiodinases and thyroid hormone receptors that begin by 7-8 weeks' gestation, can be an evidence that thyroid hormone is important in fetal neurodevelopment.15About 10-15% of pregnant women have thyroid dysfunction during the first half of pregnancy.16-18Thyroid dysfunction specially mild or subclinical hypothyroidism is associated with impaired neurodevelopment in the offspring and there is a link between this problem and mental retardation in the newborn.19Sixty one diabetic pregnant women were enrolled in this study, 22 patients (36%) were diagnosed as GDM and 39 patients (64%) had pre-gestational diabetes. There were 35 healthy pregnant women as control group. Results showed that 25.6% of pregnant women with pre-gestational diabetes and 4.5% of women with GDM and 8.6% of control group had thyroid dysfunction. That sub clinical hypothyroidism was more prevalent. Thyroid dysfunction in GDM patients had no significant difference with control group. In Luisa Ruas et al study the prevalence of thyroid dysfunction in GDM was 5.1% that is comparable with our study.20 Agarwal et al study reported that thyroid dysfunction rate is similar in GDM and non-diabetic pregnant women,9 but Velkoska Nakova et al reported higher prevalence of hypothyroxinemia in GDM than non-diabetic controls.10A study in normal pregnant women in Iran reported thyroid dysfunction prevalence as 2.6% that is similar to our study.7 and 40.9%8 respectively; that is higher than our study. The cause of this difference may be because most of our patients had type 2 diabetes (80%) rather than type 1 diabetes (20%), whereas in Gllas PR and Lois Jovanic P study all patients had type 1 diabetes and thyroid dysfunction is higher in this group.7,8 In another study in non-pregnant diabetic women in Iran the prevalence of thyroid dysfunction was 40% in type 1 and about 20% in type 2 DM.21,22In Gllas PR and Lois Jovanic P studies thyroid dysfunction in pregnant women with type 1 diabetes was 40%23 However, Di Gilio AR and colleagues study showed no significant difference between TSH, FT4, FT3 levels in diabetic and healthy pregnant women.24 In this study sample size was 15 women with type 1 diabetes and 13 healthy persons, which was small and may affect the results. Gllas PR also showed the same frequency of clinical hypothyroidism as our study.7Gonzalez et al. also showed that 26% of pregnant women with type 1 diabetes and 4% of healthy pregnant women had thyroid dysfunction which is comparable with our study.16,17 Some studies showed that even without the overt thyroid dysfunction, autoimmune thyroid disease increases abortion rates up to 3 to 5 times.14Antithyroid antibodies can cross the placenta and therefore they can influence fetal brain and neurodevelopment. In the present study, positive TPO Ab was seen in 27% of women with GDM and in 36% of pregnant women with pre-gestational diabetes and 23% of control group and there was no statistically significant difference between them.25 Luisa R et al study evaluated Tg Ab and Anti TPO Ab in 408 GDM women and reported high titer of antibodies in 5% of study population.19 The difference between mentioned studies and our study could be due to regional differences, race and iodine consumption. Because of consumption of iodine salt for correction of iodine deficiency in our country, it can increase thyroid auto-immunity and rise in thyroid auto antibodies titer.14 In Ortega study just TPO Ab > 251 U / ml (strongly positive) was considered, whereas in our study all patients with positive TPO Ab, have been considered. Bech et al also reported 17.85% positive titer of anti-microsomal antibody in a group of type 1 diabetes pregnant women.26Ortega-Gonzalez et al studied 50 healthy pregnant women, 50 GDM women and 50 pregnant women with type 2 diabetes TPO Ab> 251U/ml ( strongly positive) was found in 10% of healthy women, 10% of pregnant women with type 2 diabetes and 6% in women with GDM without statistically significant difference.Positive titer of auto antibody in pregnancy may be predictive of higher prevalence of post-partum thyroiditis in our population. Therefore for long term prospective studies to follow up of patients with positive antibodies and subclinical Thyroid dysfunction after delivery is recommended.This study showed that thyroid dysfunction has a high prevalence in women with pre-gestational DM and therefore thyroid function should be evaluated in these patients during pregnancy. Thyroid dysfunction was equally prevalent in women with GDM and normal pregnant women but 27% of them had positive titer of TPO Ab that warrants follow up for post-partum thyroiditis, dysfunction.


An analysis of cohorts of patients with compensated cirrhosis caused by the Hepatitis B Virus (HBV) indicates that the disease decompensates at an annual rate of 1.5% to 5%. Following decompensation, the 5-year survival rate has varied from 14% to 35% . Given i


Streptococcus pneumoniae strains has caused significant health problems worldwide.The development of antibiotic resistance among Streptococcus pneumoniae strains isolated from clinical specimens.The aim of this study was to determine antibiotic resistance pattern and serotypes distribution of Streptococcus pneumoniae strains were isolated from Tehran Hospital’s laboratory from 2008 to 2012. Antimicrobial susceptibility testing was performed using broth microdilution method and minimum inhibitory concentration (MIC) of each strain was determined. to verify the resistant strains and demonstrate the presence of antibiotic resistant genes, the PCR was performed.A total of fifty The study showed that three strains (6%) and six strains (12%) indicated intermediate resistance and complete resistance to penicillin, respectively, 58% strains were susceptible to ceftazidime, two ones (4%) indicated resistance to ciprofloxacin, one (2%) indicated intermediate resistance to ceftriaxone , two strains (4%) indicated complete resistance and four (8%) strains indicated resistance to vancomycin.Streptococcus pneumoniae strains with multiple resistance needs permanent monitoring of antibiotic susceptibility patterns of clinical isolates. We have found that ceftazidime is not a suitable drug for choosing the treatment of pneumococcal infections.The emergence of  Streptococcus pneumoniae (S. pneumoniae) is a major invasive pathogen that is the most common cause of community-acquired pneumonia (CAP)   (ia (CAP) . Despiteia (CAP) , 3. Baseia (CAP) , 5. Thisia (CAP) . Based oerotypes , 7. In aerotypes . In thes created , 10. Recsistance . Howeversistance , 12. In ountries . In one  average . Another average . Moreove average . In addibiotics) , 18.S. pneumoniae strains with multiple-resistance needs permanent monitoring of antibiotic susceptibility patterns of clinical specimens. The aim of this study was to determine the antibiotic resistance pattern and measurement of minimum inhibitory concentration (MIC), of the serotypes determined, S. pneumoniae strains isolated from clinical specimens.As a consequence, the emergence of Antimicrobial agents used in this study consist of penicillin G  , Ciprofloxacin (200 mg/100mL) , ceftazidime  , ceftriaxone  , vacomycin  . Mueller-Hinton broth , defibrinated sheep blood , 96-well microplate (from Technogen) and injectable distilled water.S. pneumoniae strains were isolated from Tehran Hospital’s laboratory from 2008 to 2012. In a previous study, all isolated strains were identified and their serotypes were determined and classified. They were then lyophilized and stored at -80oC.A total of fifty In this study, based on CLSI and EUCAST criteria  , 20  andDesigning of the 96-well microplate was performed as follows, for preparation of the serial dilutions of selected antibiotics, a stock solution of each antibiotic was prepared. In a 96-well microplate; one row for negative control, one row for positive control and three rows for tests were marked, respectively. Thus, each 96-well microplate was considered to measure the antibiotic sensitivity of two bacterial strains. The negative control rows for each case consist of the serial dilutions of the antibiotic, only culture media (Mueller-Hinton broth containing 3% sheep blood) and lacking bacterial suspension. The positive control rows consist of culture media (Mueller-Hinton broth containing 3% sheep blood) and the bacterial suspension. The positive control rows lack an antibiotic. The three rows of tests with equal conditions were used for each bacterial strain testing. The test rows composed of serial dilutions of antibiotics, culture media and bacterial suspension.The stock solutions of each antibiotic were prepared. Different concentrations of antibiotic were then prepared and loaded into defined rows of 96 well microplates from the base antibiotic stock solution. Plates were incubated at 35°C for 24 hours until evaporation and finalization of the process of loading the antibiotic. The plates were stored in a refrigerator set at 4°C whenever required. In this study, ten serial dilutions were defined for penicillin. The stock was prepared from penicillin G  so that each 1µL volume of stock contained 40 µg of antibiotic. Seven serial dilutions were defined for ciprofloxacin. The stock was prepared from ciprofloxacin  so that each 1µL volume of stock contained 2µg of antibiotic. Ten serial dilutions were defined for ceftazidime. The stock was prepared from ceftazidime  so that each 1µL volume of stock contained 40 µg of antibiotic. The stock was prepared from ceftriaxone  so that each 1µL volume of stock contained 10µg of antibiotic. Eight serial dilutions were defined for ceftriaxone. Finally nine serial dilutions were defined for vancomycine. The stock was prepared from the vancomycin vial (500mg) that each 1µL volume of stock contained 20 µg of antibiotic. The details of the serial dilutions prepared for each antibiotic are presented in oC. When the absorption rate of bacterial suspension was equivalent to McFarland 0.5 turbidity standard in 620 Nanometer wavelengths, it was used as the bacterial inoculums.Based on CLSI standard, using the broth microdilution method, antimicrobial susceptibility testing was carried out. For this purpose, the turbidity of bacterial suspension equivalent to McFarland 0.5 turbidity standard which represents 1.5×108 bacteria/mL was prepared. A colony of each bacterium was inoculated into a BHI broth medium containing 3% sheep blood separately, and incubated at 37As aforementioned, each antibiotic was loaded in triplicates row and employed for each serial dilution testing in the 96-well microplate. In the same manner, one row of the 96-well microplate was considered as a negative control for each antibiotic and one row of the 96-well microplate was considered as a positive control which consists of culture medium and bacterial suspension. A volume of 150µL Mueller-Hinton broth containing 3% sheep blood was added to each well and was shacked for 30 seconds. Finally, 50µL of bacterial inoculums with McFarland 0.5 turbidity standard were added to wells.In this study, a daily maximum of four bacterial strains were tested. For each case, the pre- loaded plates were removed from the refrigerator and 150 μL of Mueller-Hinton broth containing 3% sheep blood was added to each well. The negative control well or blank did not contain bacterial inoculums, whereas the positive control wells were free from antibiotics. The McFarland 0.5 standard provides turbidity (OD = 7) comparable to that of a bacterial suspension containing 1.5 ×108 colony-forming units (CFU)/mL or 1.5 × 105 CFU/μL. The turbidity of the prepared bacterial suspensions was compared by observing the black lines through the suspension. A 50μL volume of this suspension was added to each well to obtain a final concentration of inoculums, which was approximately 50 × 1.5 × 105 CFU/wells. Two bacterial strains were tested in a plate separately. Inoculated plates were incubated at 37°C, and the optical density (OD) of each well was measured at time 0 and 24 hours after initiation of the incubation using an ELISA reader device set at 450 Nanometer. The mean of the OD of different concentrations of antibiotics per well for each bacterium was compared and analyzed using unilateral variance analysis (ANOVA). In our statistical analyses, α = 0.005 was considered acceptable significant variation and the results were analyzed using SPSS Ver. 16.S. pneumoniae. In our study, the mentioned primers pair were selected and synthesized by CinnaGen Co Iran. In purpose of detection and amplification of pbp2b gene, overnight growth bacterial cells of an exponential phase were subjected to DNA extraction using the salting out method , and 20 (isolated from throat) had fully resistance. Furthermore, serotypes 17 (isolated from blood) and 20 (isolated from throat) have MICs more than 10µg/mL. The results of molecular detection gene pbp2b which amplified fragment 1500 bp showed that 4 strains revealed the fragment 1500 bp   in clinical samples . In anoticillin) . Meanwhiicillin) . In Chincrobials . Moreovene 14.3% . Resultsne 14.3% . So, forbacteria . This mebacteria . Finally


In an attempt to find an early, clinically relevant metabolic signal of modest IAH, we investigated abdominal wall metabolite concentrations in a small group of patients undergoing laparoscopic surgery. We hypothesized that elevated intra-abdominal pressure [IAP] due to pneumoperitoneum leads to an increased lactate/pyruvate [L/P] ratio Six patients scheduled for elective laparoscopic gastric fundoplication were studied. Two hours before surgery, a microdialysis catheter  was inserted into the RAM under local anaesthesia. Catheter placement was confirmed by ultrasound. The microdialysis perfusion rate was set at 0.3 μL/min. Dialysate was collected hourly prior to pneumoperitoneum, during pneumoperitoneum, and for 2 h after pneumoperitoneum resolution. IAP was maintained at 12 to 13 mmHg during the surgery. The glucose, glycerol, pyruvate and lactate contents of the dialysate were measured.p = 0.03). The median pneumoperitoneum duration was 86 (77 to 111) min. The L/P ratio at 2 h post-pneumoperitoneum was not different from that at baseline (p = 1.0). No changes in glycerol or glucose levels were observed.The median (interquartile range) L/P ratio was 10.3 (7.1 to 15.5) mmol/L at baseline. One hour of pneumoperitoneum increased the L/P ratio to 16.0 (13.6 to 35.3) mmol/L (IAH of 12 to 13 mmHg, even for a relatively short duration, is associated with metabolic changes in the abdominal wall muscle tissue of patients undergoing laparoscopic surgery. We suggest that tissue hypoperfusion occurs even during a modest increase in IAP, and intramuscular metabolic monitoring could therefore serve as an early warning sign of deteriorating tissue perfusion. The development of intra-abdominal hypertension [IAH] in critically ill patients admitted to the ICU is an independent predictor of mortality . The incThe diagnosis of IAH/ACS is made based on the measurement of intra-abdominal pressure [IAP], and it is dependent on the accuracy and frequency of the measurements . To dateMicrodialysis has been successfully used in animal experiments to assess metabolic changes during IAH. A high lactate-pyruvate [L/P] ratio was detected in the dialysate from the rectus abdominis muscle [RAM] in animal models of severe IAH . More geIn an attempt to find an early metabolic signal of IAH, we measured the abdominal wall metabolite concentrations in a small group of patients undergoing laparoscopic surgery. We hypothesized that IAP elevation during pneumoperitoneum leads to insufficient tissue perfusion of the RAM and that the L/P ratio is increased in parallel as a signal of anaerobic metabolism.The study was approved by the University of Tartu Ethics Review Committee on Human Research (protocol number: 170/T-11 28.04.2008). Informed consent was obtained prior to inclusion in the study. The study was performed in accordance with the Helsinki Declaration. Because this was a prospective observational study and not an interventional trial, we did not register the study with clinicaltrials.gov.Eight patients scheduled for elective laparoscopic surgery were enrolled. Two of these patients were excluded because of catheter misplacement during the surgery. All patients underwent laparoscopic gastric fundoplication. Altogether, three women and three men with a median age of 36 (30 to 42) years were studied. The patients' median body mass index was 26.2 (23.3 to 28.1), and the median haemoglobin concentration before surgery was 145 (137 to 160) g/L.2 ranging from 30% to 50%, as decided by the attending anaesthesiologist. Atracurium besylate  was used as a muscle relaxant. To ensure an equal degree of muscle relaxation in every patient, the dose was adjusted according to kinesiomyography . Ventilators were set in volume-controlled mode and regulated to achieve normal (35 to 45 mmHg) end-tidal CO2. The end-expiratory pressure was set at 0 cmH2O. Patient vital signs during anaesthesia were recorded with a GE Aisys anaesthesia machine . Blood samples for lactate measurements were drawn from an arterial cannula before and after pneumoperitoneum. Pneumoperitoneum was introduced by insufflation of CO2 at the beginning of surgery. The IAP was held at 12 to 13 mmHg using an automated insufflator.Five patients received sodium thiopental and one patient received propofol for the induction of general anaesthesia. During all surgeries, anaesthesia was maintained with sevoflurane with FiOTwo hours before surgery, a microdialysis catheter  was inserted into the RAM under local anaesthesia (2% lidocaine). Catheter placement was confirmed by ultrasound . The microdialysate perfusion rate was set at 0.3 μL/min. Dialysate was collected hourly prior to pneumoperitoneum, during pneumoperitoneum, and for 2 h after pneumoperitoneum resolution. The samples were stored in a freezer at -80°C for a maximum of 10 months at Tartu University Hospital, Estonia and were sent to Tampere University Hospital in a single shipment for further analysis. The glucose, lactate, pyruvate and glycerol contents of the microdialysates were measured with a CMA 600 analyser .p < 0.05.This was a pilot study, and prior RAM L/P ratio data for IAH in humans were not available. We were therefore not able to perform any meaningful power calculations for sample size estimation. Rather, these data will be used for those purposes in future studies to allow for adequate sample size calculation. The first analysis was undertaken after obtaining data from six patients. Statistical analysis was performed using GraphPad Prism 5.02 . The Wilcoxon matched-pairs test (non-parametric) was used to test the median differences in paired data. Data are presented as medians with interquartile ranges. Differences were considered statistically significant at p = 0.2). The median arterial pCO2 was 34.6 (32.9 to 39.9) mmHg before and 43.0 (42.0 to 45.7) mmHg after pneumoperitoneum (p = 0.003).The surgery and clinical course were uneventful in each patient. One patient experienced syncope before anaesthesia during intravenous line insertion. This resolved quickly upon the intravenous [IV] injection of 1 mg atropine. During the surgery, this patient's mean arterial pressure [MAP] was 94 (88 to 103) mmHg. The average volume of IV infusions administered during the operations was 1,600  mL. The median pneumoperitoneum duration was 86 (77 to 111) min. IAP during the surgery was stable at 12 (12 to 13) mmHg. The patient's global haemodynamics were stable. The median MAP during pneumoperitoneum was 77 (74 to 94) mmHg, and the median abdominal perfusion pressure [APP] was 65 (62 to 82) mmHg. The median duration of MAP ≤ 65 mmHg was 1.5 (0.0 to 13.8) min. The median blood lactate level was 0.7 (0.6 to 1.4) mmol/L before and 1.0 (0.6 to 1.5) mmol/L after pneumoperitoneum (p = 0.03). L/P ratio values were not correlated with MAP or APP. Individual dynamics are shown in Figure The L/P ratio increased during pneumoperitoneum in all patients. The median L/P ratio was 10.3 (7.1 to 15.5) mmol/L at baseline, and it increased to 16.0 (13.6 to 35.6) mmol/L after 1 h of pneumoperitoneum (p = 0.44), and at 80 to 120 min, the level was 0.9 (0.1 to 4.4) mmol/L (p = 0.81). The blood glucose level increased to 4.4 (2.1 to 6.1) mmol/L (p = 0.06) at 1 h after pneumoperitoneum and was 2.7 (0.02 to 6.1) mmol/L (p = 0.75) at 2 h after the pneumoperitoneum.The median RAM tissue glucose level at baseline was 3.3 (0.1 to 4.4) mmol/L  at 60 min and to 326 (144 to 730) μmol/L (p = 0.13) at 80 to 120 min. At 1 h after pneumoperitoneum, the glycerol level was 204 (84 to 284) μmol/L (p = 0.31), and at 2 h after pneumoperitoneum, the glycerol level was 155 (105 to 272) μmol/L (p = 0.63).The median glycerol level at baseline was 103 (65 to 169) μmol/L Figure . During The present study demonstrates that IAP increases to only 12 mmHg for a relatively short duration are associated with metabolic changes in the abdominal wall muscle tissue of patients undergoing laparoscopic surgery. This indicates that anaerobic metabolism, and thereby RAM hypoperfusion, occurs upon very modest increases of IAP.Several recent studies have shown that an increase in the mean IAP is associated with adverse ICU outcomes ,14-16. Hper se. Microdialysis is an invasive, relatively expensive and time-consuming procedure. Therefore, this study lacks comparative data from control patients or other anatomical regions of the same patient. Because the insertion of a central venous line was not clinically indicated in these patients, we also did not measure global indices of preload and circulation, which would be required for an in-depth interpretation of the microdialysis data. However, the observed arterial blood pressure and heart rate dynamics do not suggest remarkable alterations in cardiac output during pneumoperitoneum.The limitations of our study include its small sample size and microdialysis-related problems, specifically catheter displacement before or during surgery in two patients. The first limitation is expected, given that we are reporting these results as a pilot trial. The latter reflects the limitations of the method The L/P ratio of a tissue reflects its reaction to changing oxygen and glucose supplies , and it During ischemia, both the oxygen and the glucose supply to the tissue are compromised. Inadequate delivery combined with increased uptake leads to a marked decrease in the interstitial glucose levels during severe ischemia . We obseElevated tissue glycerol levels indicate cell membrane damage. Bäckström et al. have demonstrated that mesenteric vein glycerol is an indicator of splanchnic ischemia . We obse2 insufflation [IAH has a global impact on the human body. It results in a variable series of pathophysiological consequences, the specifics of which depend on the underlying diagnosis . Althougfflation . In all fflation . The Eurfflation .The present study demonstrates that even a mild to moderate increase of IAP for a short duration is associated with unfavourable metabolic changes in the RAM. Further investigation is required to evaluate whether microdialysis can be used as a diagnostic tool for the detection and grading of IAH.ACS: abdominal compartment syndrome; APP: abdominal perfusion pressure; GE: General Electric; IAH: intra-abdominal hypertension; IAP: intra-abdominal pressure; IV: intravenous; L/P ratio: lactate-to-pyruvate ratio; MAP: mean arterial pressure; RAM: rectus abdominis muscle.The authors declare that they have no competing interests.LM participated in the design of the study, data acquisition and analysis, and drafted the first version of the manuscript. JK was involved in the study design and data acquisition, and revised the manuscript. JS contributed to the concept and the design of the study, and revised the manuscript. JT contributed to the laboratory analysis, data processing and critical revision of the manuscript. All authors have read and approved the final manuscript.


Chronic refractory angina is a challenging clinical problem with limited treatment options. The results of early cardiovascular stem cell trials using ABMMC have been promising but have utilized intracoronary or intramyocardial delivery. The goal of the study was to evaluate the safety and early efficacy of autologous bone marrow derived mononuclear cells (ABMMC) delivered via percutaneous retrograde coronary sinus perfusion (PRCSP) to treat chronic refractory angina (CRA).8 ± 4.3*108 mononuclear and 1.65*107 ± 1.42*107 CD34+ cells by PRCSP..From May 2005 to October 2006, 14 patients, age 68 +/- 20 years old, with CRA and ischemic stress-induced myocardial segments assessed by SPECT received a median 8.19*10ABMMC delivery was successful in all patients with no arrhythmias, elevated cardiac enzymes or complications related to the delivery. All but one patient improved by at least one Canadian Cardiovascular Society class at 2 year follow-up compared to baseline (p < 0.001). The median baseline area of ischemic myocardium by SPECT of 38.2% was reduced to 26.5% at one year and 23.5% at two years (p = 0.001). The median rest left ventricular ejection fraction by SPECT at baseline was 31.2% and improved to 35.5% at 2 year follow up (p = 0.019).PRCSP should be considered as an alternative method of delivery for cell therapy with the ability to safely deliver large number of cells regardless of coronary anatomy, valvular disease or myocardial dysfunction. The clinical improvement in angina, myocardial perfusion and function in this phase 1 study is encouraging and needs to be confirmed in randomized placebo controlled trials. An increasing number of patients with coronary artery disease remain symptomatic with disabling angina despite the optimal use of antianginal medications and percutaneous or surgical revascularization ,2. TheraPatients with Canadian Cardiovascular Society (CCS) class III-IV angina despite maximal medical or surgical therapy who were ineligible for further percutaneous or surgical revascularization (based on coronary anatomy) and who had evidence for reversible ischemia on an exercise single photon emission computed tomography (SPECT) were enrolled. A committee comprising two cardiovascular surgeons and two interventional cardiologists determined the ineligibility for percutaneous or surgical revascularization. Patients with acute myocardial infarction, percutaneous or surgical revascularization within six months of enrollment in the study, a history of malignant disease, severe renal dysfunction, or unexplained hematology or biochemical abnormalities were excluded. The local ethics committee at Clinica Maison de Sante and Centro Cardiovascular San Nicolá s approved the protocol and all patients gave informed consent. The review and analysis of the data was also approved by the IRB at the University of Utah.The baseline screening assessment of patients included clinical evaluation, electrocardiogram (ECG), laboratory evaluation . Patients kept a record of daily angina frequency for three weeks, and the severity of angina was graded according to the CCS class at baseline, 3, 12, and 24 months. Within two weeks prior to cell therapy, exercise capacity was evaluated using bicycle ergometry in conjunction with SPECT imaging to assess myocardial ischemia and left ventricular (LV) function.Patients had complete blood count, creatine kinase levels and ECG performed immediately prior to and immediate after the procedure. Complete blood count, creatine kinase, and troponin T levels were also assessed eight hours post procedure with an ECG prior to discharge. All patients were monitored in the cardiac intensive care unit for 12 hours after the procedure and were discharged the following day. A transthoracic echocardiogram was performed prior to discharge to detect the presence of pericardial effusion. The clinical follow-up evaluations including the laboratory tests described above were performed 3, 12 and 24 months after the injection procedure. At 12 and 24 months of follow-up exercise bicycle testing, and gated SPECT were performed.All patients performed a symptom-limited bicycle exercise test with a 20-W starting load and 10-W increments per minute at baseline and at 12 and 24 months after the injection procedure. Antianginal medication was continued. The test points were angina pectoris, physical exhaustion, dyspnea, significant decrease in blood pressure (10 mmHg), or achievement of maximal age-related heart rate. A 12-lead ECG was recorded before, during, and after the test. The total exercise duration, maximal workload achieved in percentages  at baseline was compared to 12 and 24 months after ABMMC delivery.99 mTechnetium Sestamibi injection using a double-head SPECT camera as previously described.  Using the bicycle exercise test, 99 mTechnetium Sestamibi was injected intravenously at peak exercise (85% of the estimated heart rate) which was continued for 2 minutes after tracer injection and images were obtained 30 minutes after the stress test. Reconstruction yielded long-and short -axis projections perpendicular to the heart axis. The short-axis slices were displayed in polar map format.For the SPECT examination, a one-day rest-stress protocol was used. On the same day, images at rest were obtained 60 minutes after the 8 ± 4.3*108 mononuclear cells and 1.65*107 ± 1.42*107 CD34+ cells.A volume of 300 ml of bone marrow was harvested from the iliac crest under local anesthesia and placed in heparinized Hanks' balanced salt solution. ABMMC were isolated by Hess 6% density gradient centrifugation with a final suspension volume of 50 ml and a mean cell concentration of 8.19*10The femoral vein was cannulated with a 7 French sheath, a 6 French catheter was placed in the coronary sinus and a 0.035 mm hydrophilic guide wire was placed in the interventricular or lateral vein followed by placement of a peripheral balloon into the mid portion of the coronary sinus to allow nonselective delivery of cells. . The balloon was inflated at very low pressure 1 to 2 atm) for 10 minutes producing stagnation of the flow as previously described with infusion of the ABMMC. 50 mls of ABMMC were injected manually through the balloon at a rate of 10 mls per minute. The average total procedure time for cell delivery was 30 minutes. Ventricular arrhythmias were not observed during cell delivery or hospitalization and no patient had post procedural pericardial effusion by two-dimensional echocardiography before discharge. One patient died four months following the procedure due to liver cirrhosis, decompensated heart failure and pulmonary hypertension.The frequency of angina episodes per day decreased from 2.9 ± 3.9 at baseline to 1.0 ± 1.4 at 3 months (p < 0.001), 0.7 ± 1.2 at 12 months (p < 0.001) and 0.6 ± 1.2 at 24 months (p < 0.001). Angina improved in 13/14 patients and the mean Canadian Cardiovascular Society class improved from 3.2 ± 0.6 baseline to 1.9 ± 0.7 at 1 year (p < 0.001) and 1.8 ± 0.4 at 2 years (p < 0.001). The individual changes for anginal episodes per day for the 14 patients are shown in Figure At baseline, 14 patients had exercise duration time of 3.1 ± 1.4 minutes. This improved to 6.8 ± 1.0 and 7.4 ± 1.7 minutes at 12 and 24 months of follow-up respectively, (p < 0.001). The individual changes for exercise time are shown in Figure The median percent of ischemic myocardium by SPECT was 38.2% at baseline and was reduced to 26.5% and 23.5% (p = 0.001) at 12 and 24 month follow-up (Table Median rest LV ejection fraction by SPECT at baseline was 31.2% and was improved to 35.4% and 35.5% at 12 and 24 month follow-up respectively (p < 0.019). (Table 9 described a reduction in anginal symptoms, improved wall motion and improved wall thickening at 3 'months follow-up in 8 patients. In addition, the area of hypoperfused myocardium was reduced on SPECT. Fuchs et al reported a reduction in anginal symptoms and improved myocardial perfusion in 27 patients with a trend toward improved LV ejection fraction [This study demonstrates that ABMMC delivered by PRCSP in patients with CRA and stress-inducible ischemia was safe, may reduce anginal symptoms and improve exercise capacity. Stem cell transplantation by PRCSP is an alternative route of delivery to the myocardium for patients with coronary artery occlusion at their initial portions, or for patients with severe multivessel disease or occluded venous and arterial grafts. In this study we have demonstrated that, in patients with chronic ischemia with or without severely impaired LV function, that ABMMC injection enhances myocardial perfusion. In addition we observed that improved systolic LV function occurred only in patients with low ejection at baseline. Improvement and maintenance of the LV ejection fraction at 24 months follow-up was most likely a result of increased myocardial perfusion and global regional wall motion. This resulted in a reduced LV end-systolic volume. Consequently, the therapeutic effect appears to be more related to enhanced myocardial contractility rather than induction of LV reverse remodeling. The findings of the present study are consistent with the hypothesis that ABMMC promote angiogenesis, resulting in increased myocardial perfusion . The prefraction . Perin efraction . In a rafraction . In thisfraction ,22 and tABMMC: autologous bone marrow mononuclear cells; CCS: Canadian Cardiovascular Society; CRA: refractory angina; ECG: electrocardiogram; LV: left ventricular; PRCSP: percutaneous retrograde coronary sinus perfusion; SPECT: single photon emission computed tomography.The authors declare that they have no competing interests.JT, RFV, AC, JC, CC, AC, JE, CY, JC enrolled and treated the patients. JT, RFV, AP conceived the study. JT, TH, AP all help to draft the manuscript and were involved in the data analysis. All authors read and approved the final manuscript.


Pungtungia herzi were developed using a combination of next-generation sequencing and Sanger sequencing. One hundred primer sets in the flanking region of dinucleotide and trinucleotide repeat motifs were designed and tested for efficiency in polymerase chain reaction amplification. Of these primer sets, 16 new markers (16%) were successfully amplified with unambiguous polymorphic alleles in 16 individuals of Pungtungia herzi. Cross-species amplification with these markers was then examined in two related species, Pseudopungtungia nigra and Pseudopungtungia tenuicorpa. Fifteen and 11 primer pairs resulted in successful amplification in Pseudopungtungia nigra and Pseudopungtungia tenuicorpa, respectively, with various polymorphisms, ranging from one allele (monomorphic) to 11 alleles per marker. These results indicated that developing microsatellite markers for cross-amplification from a species that is abundant and phylogenetically close to the species of interest is a good alternative when tissue samples of an endangered species are insufficient to develop microsatellites.Nuclear microsatellite markers for  Pseudth Korea ,2. The parasites –5.A good understanding of the genetics of endangered species can assist in developing strategies for their conservation ; howeverPungtungia herzi) is widely distributed in most rivers and lakes in South Korea, northern China, and southern Japan [Pungtungia herzi uses two reproductive strategies, nonparasitic crevice spawning and brood parasitism with multiple host species, including Coreoperca herzi, C. kawamebari, Pseudobagrus nudicepts, and Odontobutis obscura (Perciformes: Odontobutidae) [Pungtungia herzi and Pseudopungtungia nigra appear to be closely related phylogenetically, due to their morphological similarities and rather distinct zoogeographic distributions [Pungtungia herzi and Pseudopungtungia nigra was reported in the Ungcheon River [The striped shiner  –19. Pungibutions . In factPungtungia herzi using a NGS protocol [Pungtungia herzi in the two related endemic species, Pseudopungtungia nigra and Pseudopungtungia tenuicorpa.Next-generation sequencing (NGS) methods are revolutionizing molecular ecology by simplifying the development of molecular genetic markers, including microsatellites . In thisprotocol . We then2.2.1.Pungtungia herzi. The total number of contigs was 9708 and the number of singletons was 146,093. A total of 1981 regions containing perfect dinucleotide/trinucleotide repeats were identified. The most frequent dinucleotide type in the Pungtungia herzi genome was CA repeats (51.66%), followed by AT (25.10%), CT (21.77%), and GC (1.47%). These results are consistent with previous findings that the CA repeat is the most frequent in 353 species of vertebrata genomes and is generally 2.3-fold more frequent than the second most common microsatellite, AT [We obtained 243,749 total reads (91.87 Mb) for lite, AT . Of the 2.2.Pungtungia herzi. The nucleotide sequences of the 16 new microsatellite markers were deposited in the GenBank database under accession numbers JQ889798–JX915813  produced strongly amplified polymerase chain reaction (PCR) products in all specimens with unambiguous alleles in JX915813 .Pungtungia herzi, these 16 markers were polymorphic , and PH_CT03, PH_CA37, PH_ACT01 (p < 0.05) departed from HWE  and are recommended for use in future population genetics studies of Pungtungia herzi (Hardy–Weinberg equilibrium (HWE) was tested in the Gyeongho River population. In this population, PH_CA36, PH_CA38, PH_ATC01  was amplified only in Pseudopungtungia tenuicorpa. The number of alleles for the 15 markers amplified in Pseudopungtungia nigra ranged from 1 to 11. The number of alleles on the 11 markers from Pseudopungtungia tenuicorpa ranged from 1 to 6  following the manufacturer’s protocol and stored at −20 °C. The quality of genomic DNA was checked on 1% agarose gels with a spectrophotometer . The NGS libraries were generated with aliquots of approximately 10 μg of genomic DNA from a single 3.2.The sequence reads from the 454 GS-FLX were assembled using Newbler 2.6  with a 96% minimum overlap identity. The dinucleotide and trinucleotide repeats of more than four iterations were searched using the perl program “ssr_finder.pl” ,24. Thes3.3.® version 2.0; TaKaRa Bio, Otsu, Japan) in a final reaction volume of 20 μL. The PCR amplification protocol was 94 °C for 5 min, followed by 30 cycles of 94 °C for 30 s, 56 °C for 45 s, and 72 °C for 45 s, then 12 cycles of 94 °C for 30 s, 53 °C for 45 s, and 72 °C for 45 s, and a final extension at 72 °C for 10 min. Subsequently, the PCR product was added to Hi-Di™ formamide with 500 LIZ® size standard  and run on an ABI 3730xl automatic sequencer . The microsatellite profiles were examined using GeneMarker ver. 1.85 . To verify the flanking sequences of the microsatellite repeats and to confirm whether target fragments of interest were amplified, PCR was performed with 8 pmol of forward and reverse primers without fluorescent-labeled M13 primer. The amplified products were sequenced using an ABI 3730XL and the targeted sequences were checked for correct amplification.The PCR mix contained 10 ng of genomic DNA, 8 pmol each of the reverse primer and fluorescent labeled  M13 primer, and 4 pmol of the forward primer, Premix Taq (TaKaRa Ex Taq3.4.HO and HE using Arlequin ver. 3.11 [Pungtungia herzi was analyzed and the program MICRO-CHECKER ver. 2.2.3 [We estimated the proportion of polymorphic markers and the average number of alleles per marker and calculated er. 3.11 . Hardy-Wr. 2.2.3  was used4.Pungtungia herzi. Of these, 15 and 11 markers were applied successfully in the endangered species Pseudopungtungia nigra and Pseudopungtungia tenuicorpa, respectively. This achievement demonstrates that the microsatellite markers developed using the NGS method from a related species can be applied effectively to an endangered species. Developing these microsatellite primers will assist work to determine genetic relationships among the species, as well as improving our understanding of the population genetic structure within each species.We developed 16 new microsatellite markers in


Rhipicephalus sanguineus and Ctenocephalides felis felis on dogs were conducted over periods of 226 days and 71 days respectively.Two studies evaluating the efficacy of an imidacloprid/flumethrin collar , a deltamethrin collar , a fipronil/(s)-methoprene spot-on , a dinotefuran/pyriproxyfen/permethrin spot-on  and an amitraz/fipronil/(s)-methoprene spot-on  against repeated infestations with The first study comprised 4 groups of treated dogs and one untreated control group, and the second 3 groups of treated dogs and one control group. Each group consisted of 8 dogs. All dogs were infested with ticks and fleas at regular intervals. Ticks were counted 6 h, 18 h or 48 h after infestations and fleas 24 h after infestations. Efficacies of the treatments were calculated by comparison with the untreated control groups using standard descriptive statistics.The protective 48 h tick efficacy was 97.8% to 100% for the IVP (226 days), 69.3% to 97.4% for CP1 (170 days), 99.6% to 43.4% for CP2 (35 days) and 98% to 61.4% for CP3 (35 days).The protective 18 h tick efficacy was 98% to 99.6% for the IVP (71 days), 100% to 86.5% for CP4 (29 days), 100% to 72.8% for CP4 after re-treatment (35 days) and 98.8% to 54.3% for CP5 (35 days).The protective 6 h tick efficacy was 85.6% at Day 7 and 90.1% to 97.1% from Day 14 onwards for the IVP (70 days), 92.3% to 70.7% for CP4 (35 days), 97.5% to 65.2% for CP4 after re-treatment (35 days) and 95.1% to 51.8% for CP5 (35 days).The protective 24 h flea efficacy was 99.5/90.9% to 100% for the IVP (71/226 days), 66.7% to 83% for CP1 (170 days), 100% to 88.5% for CP2 (35 days), 100% to 73.3% for CP3 (35 days), 100% to 98.7% for CP4 (35 days), 100% to 87.5% for CP4 after re-treatment (35 days) and 100% to 79.5% for CP5 (35 days).These data suggest that the long-term efficacy provided by a medicated collar that is effective, is a means to overcome the fluctuating efficacy of spot-on treatments resulting from a lack of pet owner re-treatment compliance, and consequently protect animals successfully against ectoparasites and probably vector-borne diseases. Rhipicephalus sanguineus is present on these animals in several regions and predominant in others , the expression of efficacy can also vary within the same molecule. There is a dose-time-dependency for lethal effects in pyrethroids [Repellent effects are a general property of pyrethroids. Their in-contact efficacy comprises a mixture of a very rapid lethal effect, a knock down effect and the so-called “hot-foot effect”, instead of a vapour-based classical repellent effect. Besides the well-described difference of specific ectoparasiticidal potency amongst different pyrethroids of even the same chemical subclass [e.g. Mendes  describeethroids  so that ethroids , the haiR. sanguineus, was, with the exception of Day 7 after treatment, above 90%, followed by “rapid killing” acaricidal efficacies ≥98% against the same population of ticks 12 h later -methoprene/fipronil, dinotefuran/pyriproxyfen/permethrin and (s)-methoprene/amitraz/fipronil against  on dogs . In addi on dogs .Although the effectiveness of the various remedies tested in these studies may at various stages after their application be excellent, it is the number of living ticks or fleas remaining on treated dogs, when efficacy decreases or is poor, that are important. Tick burdens exceeding 10 individuals after treatment are quite adequate for the transmission or acquisition of organisms responsible for tick-borne diseases in the field, while a few fleas remaining after ineffective treatment are capable of inducing severe signs of flea allergy dermatitis. This implies that if pet owners do not comply with the recommended time-periods between the administration of spot-on remedies serious gaps in efficacy may occur. The long-term, persistently high efficacy of the imidacloprid/flumetrin collars would appear to be an excellent counter to these eventualities.R. sanguineus remained above 97% over a period of 8 months on regularly shampooed dogs, and above 94% on dogs regularly immersed in water. Efficacy against C. felis felis remained above 90% on shampooed dogs for the 8-month duration of the study, but declined to below 90% at 6 months in the group of dogs that were regularly immersed in water [The studies above were all laboratory based, while in the field various challenges to the efficacy of the medicated collars may occur. The most obvious of these is that dogs are inevitably going to be washed or shampooed, or swim or go out into the rain while being walked by their owners or working in the field. Importantly collars do not have to be removed during any of these events , and thain water .Rhipicephalus sanguineus and Ctenocephalides felis felis on dogs, provides the wherewithal to overcome the fluctuating medium-term efficacy of spot-on treatments resulting from a lack of pet owner re-treatment compliance. The sustained high level of efficacy of the collars against ticks 6 hours and fleas 24 hours after infestation, may well interfere with the transmission of vector-borne diseases.The 8-month long period of efficacy of medicated collars incorporating a combination of 10% imidacloprid and 4.5% flumethrin against repeated infestations of These clinical studies were completely funded by Bayer Animal Health GmbH, Monheim, Germany, of which D. Stanneck (Germany) is an employee. ClinVet is an independent Contract Development Organisation, which was contracted to manage the conduct of both studies. I.G. Horak is a long-term, contract employee of Clinvet and an Extraordinary Professor at the Universities of the Free State and Pretoria. All authors voluntarily publish this article and have no personal interest in these studies other than publishing the scientific findings that they have been involved in via planning, setting-up, monitoring and conducting the investigations and analysing the results.DS and JJF designed the study and drafted the protocol and JJF conducted the study. IGH compiled and analysed the data and wrote the initial drafts of the manuscript. All authors read and approved the final manuscript.


Exposure to dust or fumes can cause a verity of lung problems, including chronic airflow obstruction.The study was carried out on the 256 workers exposed to dust at the rubber factory. 16 workers with moderate exposure, 240 Workers with high exposure, spirometry  were performed. Information on occupational history, duration of exposure, smoking habits, alcohol consumption, respiratory symptoms  and self reported symptoms with disease were collected. By employing multiple linear regression modeling the potentially confounding effects of age, sex and body mass index were also incorporated into the analysis. Odds ratio were calculated for FVC<80% predicted in different exposure subgroups.Statistically, significant reduction in FVC, FEV1 and PEFR were found when compared to age, small airway obstruction, and also in shortness of breathing. Small airway obstructions were found in dust fume (27.2%), Smoking (30.3%), Alcohol (29.3%). Lung function indices were found to be reduced with increasing duration of exposure to working environment. The FVC of the workers exposed to factory with a mean of 3.6 ± 0.6. The FEV1, for workers exposed with a mean of 2.4 ± 0.6. The mean value of the ratio of FEV1/FVC in exposed workers was 76.8 ± 8.2 there was no statistical difference between these two means.Due to high ambient dust concentration and the observed adverse effects on lung functions worker exposed to dust have more respiratory symptoms and a grater risk of airflow obstruction. A reduction of dust exposure and secondary preventive measure is advised.


Background: Exposure to wildfire smoke has been associated with cardiopulmonary health impacts. Climate change will increase the severity and frequency of smoke events, suggesting a need for enhanced public health protection. Forecasts of smoke exposure can facilitate public health responses.Objectives: We evaluated the utility of a wildfire smoke forecasting system (BlueSky) for public health protection by comparing its forecasts with observations and assessing their associations with population-level indicators of respiratory health in British Columbia, Canada.2.5 forecasts with PM2.5 measurements from air quality monitors, and BlueSky smoke plume forecasts with plume tracings from National Oceanic and Atmospheric Administration Hazard Mapping System remote sensing data. Daily counts of the asthma drug salbutamol sulfate dispensations and asthma-related physician visits were aggregated for each geographic local health area (LHA). Daily continuous measures of PM2.5 and binary measures of smoke plume presence, either forecasted or observed, were assigned to each LHA. Poisson regression was used to estimate the association between exposure measures and health indicators.Methods: We compared BlueSky PM3 increase in BlueSky PM2.5 was associated with an 8% increase in salbutamol dispensations and a 5% increase in asthma-related physician visits. BlueSky plume coverage was associated with 5% and 6% increases in the two health indicators, respectively. The effects were similar for observed smoke, and generally stronger in very smoky areas.Results: We found modest agreement between forecasts and observations, which was improved during intense fire periods. A 30-μg/mConclusions: BlueSky forecasts showed modest agreement with retrospective measures of smoke and were predictive of respiratory health indicators, suggesting they can provide useful information for public health protection.http://dx.doi.org/10.1289/ehp.1306768Citation: Yao J, Brauer M, Henderson SB. 2013. Evaluation of a wildfire smoke forecasting system as a tool for public health protection. Environ Health Perspect 121:1142–1147;  As the global climate continues to change, more frequent and intense wildfire events and longer wildfire seasons are expected . Wildfir2.5 (particulate matter ≤ 2.5 μm in aerodynamic diameter) has been the most consistently elevated and widely measured exposure metric  Hazard Mapping System  whether forecasts are accurate and precise compared with reference measurements, and b) whether forecasts are associated with population health responses. Most evaluations of forecasting models address only the first question, but for an exposure without a “gold standard” reference measurement, like wildfire smoke, answering the second question is also important. Here we address both questions in an integrated evaluation of the operational BlueSky Western Canada Wildfire Smoke Forecasting Framework .Forecasts have been implemented for many health hazards, including extreme heat , pollen 2.5 concentrations from wildfires up to 60 hr in advance since 2010. Detailed information about the system is described elsewhere  in Canada, which is divided into 89 local health areas (LHAs) for administrative purposes . Although BlueSky produces PM2.5 forecasts up to 60 hr in advance, we present results only for the 48-hr forecasts because this is a relevant averaging period from the public health perspective.2.5. Hourly PM2.5 measurements from 36 monitoring stations  in BC  with daily mean PM2.5 values > 0.BlueSky plumes. Daily smoke plume shapes were derived from the outline of all BlueSky PMhttp://www.firedetect.noaa.gov/viewer.html). These plumes were hand-drawn by trained NOAA analysts based on imagery from seven satellites  to estimate population-weighted exposures. One DA typically includes 400–700 people  or nearest to (monitoring stations) the DA centroids. We also overlaid BlueSky Plumes and HMS Plumes with the DA centroids, and LHAs with > 50% of the total population covered by the smoke plumes were assigned a value of 1, and the other LHAs were assigned a value of 0. As a result, two continuous variables (BlueSky PM2.5 and Monitor PM2.5) and two binary variables (BlueSky Plumes and HMS Plumes) were created.0 people , and eacStatistical analyses. Four model evaluation statistics were calculated for the relationship between BlueSky PM2.5 and Monitor PM2.5: a) the Pearson’s correlation coefficient (r); b) normalized root mean squared error (NRMSE); c) index of agreement ; and d) fractional bias . These statistics were calculated in three different analyses:A global analysis, in which all forecasted and measured values at any time and location were includedA spatial-only analysis, in which forecasted and measured values were compared at fixed times for all locationsA temporal-only analysis, in which whole time series of forecasted and measured values were compared for fixed locations.BlueSky ∩ AHMS, area covered by both BlueSky and HMS plumes) and union  of the two plumes for each day (Equation 1):To quantitatively assess the extent of agreement between BlueSky and HMS plumes, the figure of merit in space (FMS)  was calcBlueSky ∩ AHMS/ ABlueSky ∪ AHMS × 100%. [1]FMS = A3 increase in PM2.5, which was equivalent to 2 SDs in the daily PM2.5 concentrations measured across all monitoring stations. For the binary variables, the effect of being covered by the smoke plume was compared with not being covered. To account for potential autocorrelation within the time-series data from any individual LHA, parameters in the regression models were calculated with generalized estimation equations (GEEs) in R , assuming an exchangeable correlation structure . Model estimates were adjusted for daily maximum temperature from the closest monitor to each LHA, day of week, holidays, and the week of the study period. A lag of 0–1 days was used for all analyses, based on the best fitted lag time for the acute effects from forest fire smoke in previous BC fire smoke studies , and less smoky areas were defined as other LHAs.2.5 versus Monitor PM2.5BlueSky PM. Model evaluation statistics for BlueSky PM2.5 forecasts compared with monitored PM2.5 observations showed modest agreement  concentration was 10.0 ± 14.5 μg/m3, with an interquartile range of 3.0–10.1 μg/m3. The mean BlueSky PM2.5 forecast ranged from 0 to 988 μg/m3 across the LHAs. The arithmetic mean was 4.0 ± 27.3 μg/m3 with an interquartile range of 0–0.1 μg/m3. The distribution of BlueSky PM2.5 was highly skewed to the right because of the large number of zero values in the output. This distribution is typical for air quality model outputs  for a 30-μg/m3 increase in Monitor PM2.5  for Monitor PM2.5. Larger point estimates and wider CIs were observed for the Monitor PM2.5 compared with the BlueSky PM2.5, partially due to some very high concentrations (up to 988 μg/m3) forecast by BlueSky. The difference between the two was attenuated when BlueSky PM2.5 estimates > 300 μg/m3 were truncated to 300 μg/m3 (The rate ratios [RRs (95% CIs)] for salbutamol dispensations were 1.08  for a 30-μg/mor PM2.5 . The RRs00 μg/m3 . In the 00 μg/m3 .BlueSky Plumes versus HMS Plumes. The mean areas of BlueSky Plumes and HMS Plumes during the study period were 153,200 and 334,500 km2, respectively. BlueSky generally forecasted smaller smoke plumes than those observed by HMS during major fire events. The mean FMS score was 0.21, with a range of 0–0.52. Higher FMS scores were observed during the major fire event periods ( periods .The number of days with ≥ 50% of the population covered by HMS Plumes ranged from 4 to 21 (of 35 days) across the LHAs, with a mean of 12 days. The mean number of days with ≥ 50% of the population covered by BlueSky Plumes was 9 (of 33), with a range of 2 to 21 days. Although the study period was 35 days, 2 days of BlueSky forecasts were missingThe RRs for salbutamol dispensations associated with BlueSky and HMS Plumes were very similar, both with a point estimate of 1.05 . The RR a) comparing its output with observations from other air quality assessment tools, and b) evaluating associations between its output and health indicators known to be associated with those other air quality assessment tools. During short-term air pollution episodes, such as wildfire smoke events, different strategies (ranging from public education to community evacuation) may be implemented based on assessment of exposure levels and their corresponding health risks. BlueSky is one of the many tools available for smoke exposure assessment, but it is different from the other tools because it provides a forecast rather than an observation in near–real-time or a retrospective measure. This study helps to highlight the potential role of smoke forecasting systems in the public health response process.Here we assess a smoke forecasting system for public health protection by 2.5 and Monitor PM2.5, with a global correlation of 0.4. The results compared well with correlations of 0.3 and 0.5 between forecasts from the two branches of the European Fire Assimilation System and the MODIS (Moderate Resolution Imaging Spectroradiometer) PM2.5 observations  increase in salbutamol dispensations in fire-affected populations of BC during the 2003 to 2010 fire seasons, using meta-regression from different LHAs. 3 increase in monitor-observed PM10 was associated with a 16% increase in the odds of an asthma-specific physician visit in a cohort of 280,000 people during the 2003 wildfire season in BC. Although 10 instead of PM2.5, the results are comparable because PM2.5 is the major fraction of PM10 from wildfire smoke  was associated with a 2% increase of odds of asthma-specific physician visits. 3 increase in PM2.5 was associated with a 16% increase in asthma hospital admissions in Los Angeles, California.We found 8% and 12% increases in salbutamol dispensations associated with 30-μg/mre smoke . In that2.5 forecasts were consistently associated with health indicators and the effect estimates were comparable with Monitor PM2.5 measurements, the estimated effects were generally smaller and the CIs were narrower. An important contributor to this result was the large range of values in the BlueSky forecasts. When forecasts > 300 μg/m3 were truncated to 300 μg/m3 (approximately double the highest Monitor PM2.5 concentration), the point estimates were larger and the CIs were wider than those calculated with the original data (Although BlueSky PMnal data .We also found a 5% increase in salbutamol dispensations associated with being covered by BlueSky or HMS plumes. For asthma-related physician visits, the increases were 6% and 9% for BlueSky and HMS Plumes, respectively. In the Larger point estimates of association were found for salbutamol dispensations in more smoky areas compared with less smoky areas, but this was not observed for asthma-related physician visits. This may suggest the increase in dispensations was more relevant to wildfire smoke exposures. Although only a few wildfire smoke studies  have repWe found that agreement between BlueSky forecasts and observed data was reasonable compared with evaluations of other existing smoke forecasting systems. Better agreement was generally observed during intense fire periods. We also found significant associations between BlueSky forecasts and respiratory health outcomes, with risk estimates consistent with those calculated using the observed data and with those reported by other epidemiologic studies. These results suggest that BlueSky forecasts can provide useful information for public health decision making. Because the 2010 fire season was among the most extreme in the history of British Columbia, ongoing evaluation during typical fire seasons is needed to further validate the role of the BlueSky forecasts in public health protection.


Charcot Neuroarthropathy (CN) is a major foot complication of diabetes which may lead to debilitating foot deformity, recurrent ulceration and lower limb amputation. Podiatrists play a key role along the continuum of the disease from diagnosis to treatment to long-term follow-up and monitoring. The aim of this clinical audit was to gather data on patients managed with active CN in a tertiary hospital podiatry department.Thirty cases (26 patients) were diagnosed with active CN during the four-year period January 2005 to December 2008, and managed through the Royal Perth Hospital Podiatry Department. Data was retrospectively collected from their medical records including demographics, diabetes parameters, CN treatment and outcomes at twelve months.Seventy three percent of cases were male. Average age 55 ± 8.6 years. Sixty percent had Type 2 insulin-requiring diabetes. Diabetes duration greater than 10 years in 83.3% cases. Diabetes control was poor (HbA1c 9 ± 1.9%). The most common locations of CN were the tarsometatarsal joints and midfoot. Seventeen percent of cases were re-activated CN. Although 40% of cases were accurately diagnosed within 4 weeks, 33% took over 2 months. Overall CN management time was 44.7 ± 25.3 weeks. Of cases (67%) which went into a total contact cast (TCC), the overall management time was 39.3 ± 29.6 weeks versus 57.5 ± 33.6 weeks for those patients who never received TCC as part of their management. Forty seven percent developed a foot ulcer within 12 months following stabilisation of their CN, and one case went on to trans-tibial amputation.These results provide an estimate for patients and practitioners as to the overall treatment time of active CN. Our findings suggest that management of CN can be prolonged, but overall treatment is reduced through the use of TCC. Despite increased awareness and education, disparity still exists for rapid diagnosis of CN. Patients are at risk of CN related foot ulceration despite close monitoring in a tertiary hospital, however, are at a relatively low risk of amputation on the short-term. The audit highlights CN is a potentially recurrent condition and patients require lifelong care and monitoring.


Methods. Unsensitized  adult primary KTRs who received a living (83%) or deceased (17%) KT in our Institution during 2004/2005 were included. DSA testing was performed monthly, and Tregs were quantified by flow cytometry every 3 months, during the 1st year after KT. All patients received triple drug immunosuppressive therapy (CNI + MMF or AZA + PDN); 83% received anti-CD25. Results. 53 KTRs were included; 32% developed DSA during the 1st year after KT. Significantly lower 7-year graft survival was observed in those who developed DSA. No difference was observed in Treg numbers up to 9 months after KT, between DSA positive and negative. However, at 12 months after KT, DSA-negative patients had significantly higher numbers of Treg. Conclusions. Early development of DSA was not associated to variations in Treg abundance. The differences in Treg numbers observed at the late time point may reflect better immune acceptance of the graft and may be associated to long-term effects. Additional inhibitory mechanisms participating earlier in DSA development after KT deserve to be sought.The aim of this study was to determine whether the abundance of regulatory T cells (Tregs) (CD4 Effective immunosuppressive regimens have greatly improved the early survival of renal allografts. However, the rate of late allograft loss has remained relatively constant . This isThe presence of donor-specific antibodies (DSAs) directed against human leukocyte antigens (HLAs) has been associated in a growing number of reports to poor prognosis of renal allografts –7. The ade novo produced antibodies against an implanted graft do not cause immediate failure . Th. Thhigh Acute graft dysfunction was defined as an unexplained increase ≥ 25% in serum creatinine (SCr). In all cases, graft biopsy was performed and urinary obstruction and/or infection were ruled out. Acute rejection was defined according to Banff ′97 and the ′03 update working classification of renal allograft pathology , 19. TreData required for glomerular filtration rate estimation (eGFR) according to the Modification of Diet in Renal Disease (MDRD) formula, at 1, 12 months, and last followup after KT was prospectively gathered from medical records.t or Mann-Whitney U tests according to data distribution . We used Chi-square test for categorical variables. The Fisher exact test was used when expected values were under 5. The association between HLA mismatches and DSA status was evaluated with Chi-Square for trend. To compare all measurements of Treg during 1 year, we used Kruskal-Wallis test. Graft survival was analyzed using the Kaplan-Meier method with log rank test for comparison of survival curves. A P < 0.05 value was considered statistically significant.For continuous variables, differences between two groups were evaluated by independent sample Student's n = 2) and followup in another institution (n = 7) during the 1st year after transplant. Therefore, the total number of KTRs included in this analysis was 53. Seventy-six first kidney transplants were performed in our center from January 2004 to December 2005. Fourteen out of 76 KTRs had a pretransplant PRA > 10% and therefore were not included. Sixty-two patients were enrolled in the study, but 9 were not considered in the final analysis. Reasons for excluding these patients were as follows: death during the first 12 months (Baseline characteristics of the patients are shown in P = 0.021). Graft survival was also lower in patients who developed class I DSA (P = 0.079). No difference was associated to development of class II DSA (P = 0.494) (data not shown).Seventeen patients (32%) developed DSA during the first year after KT. Class I DSA, class II DSA, or both were detected in 11, 4, and 2 patients, respectively. As shown in ss I DSA ; howeverυL) and as percentage of CD4+ T cells. The median (interquartile range) absolute number detected at baseline and during months 3, 6, 9, and 12 were 6.45 (1.7–10.9), 1.49 (0.2–5.0), 5.98 (2.3–11.9), 5.51 (3.9–8.6), 4.81 (2.1–14.6), respectively (P < 0.001); these correspond to 1.20% (0.4–1.7), 0.16% (0.02–0.52), 0.97% (0.34–1.44), 0.80% (0.41–1.69), and 0.68% (0.29–1.84), respectively (P < 0.001). There was a decrease in Treg number at 3 months. This was probably associated to the effect of anti-CD25 induction therapy. At month 6, the abundance of peripheral blood Tregs tended to increase and remained similar to the baseline number up to the end of the followup (12 months).Tregs were considered as absolute number (cells/P = 0.023). Between month 6 and the end of the first year, the numbers of peripheral blood Tregs differed between the patients who developed DSA and those who did not. A progressive decrease in Treg numbers was observed in patients who developed DSA. In contrast, an increase in this population was observed in patients who remained DSA negative Figures . ImportaP = 0.01).The number of biopsy-proven acute rejection episodes that occurred during the entire followup was 11. Five grafts were lost during followup. The causes of graft loss in these patients corresponded to biopsy-confirmed chronic antibody-mediated rejection (1), grade III chronic allograft nephropathy (1), grade III acute and chronic antibody mediated rejection plus thrombotic microangiopathy (1), acute cellular rejection 1B superimposed to a chronic vascular rejection (1), and acute and chronic cellular mediated rejection (1). It is worth mentioning that 4 out of these 5 patients belonged to the group that developed DSA during the first 12 months after transplantation . PatientOne of the most pursued aspects of the surveillance of these patients was the evolution of renal function for at least 5 years after transplantation. Overall, graft function assessed through SCr and MDRD eGFR shows that regardless DSA status during the first year, there was an increase in SCr as time elapsed. Albeit not significant, deterioration of graft function was more evident in the DSA-positive group according to this parameter . When we+CD25high) at different time points during the first year. We found that, as expected, Treg numbers dropped following administration of anti-CD25, but their numbers recovered 6 months after transplantation. Moreover, we observed that, in a subset of the patients, Treg numbers remained stable and tended to increase towards the end of the first year. These patients remained DSA negative, and their renal allograft had a better outcome. We also detected a group of patients whose Treg numbers decreased in the second semester after transplantation. DSA development was associated with this phenomenon. We studied prospectively a cohort of DSA-negative renal transplant patients and quantified the numbers of peripheral blood regulatory T cells , are generated in the thymus, and display a powerful suppressive capacity [One of the aims of our study was to explore the behavior of the number of Tregs during the first year after transplantation in nonsensitized kidney transplant recipients, and the temporal relationship between Treg numbers and capacity . Tregs acapacity . +CD25high T cells at month 3 after transplant. This finding is in agreement to the data published by Segundo et al. [+ and FoxP3− CD25+ T cells [+CD25high recovered by month 6 and remained stable during the first year after transplant. We were able to detect a drop in the number of CD4o et al.  and mosto et al. , and Blu T cells . In the +CD25high T cells between DSA positive/negative groups was significant at month 12. It is interesting to note that more DSA-positive patients received induction therapy with anti-CD25 compared to DSA-negative patients (94% versus 72%). Even though the difference did not reach statistical significance, the trend suggests that administration of anti-CD25 therapy might in some cases cause a prolonged decrease in Treg numbers. No differences in Treg numbers were evident during the first 6 months between the patients who developed or not DSA Figures . Howeverhigh. Since CD25 is a cell activation marker, CD25+ cells may represent activated T cells. Other markers, in particular FoxP3, are more specific for Tregs. Nevertheless, we believe that we did not overestimate the number of Tregs because we gated in CD25high cells that were virtually all FoxP3+ . This represents a weakness of the study. Another weakness of the study is that Tregs were defined as CD25l FoxP3+  and beca+CD25high cells display a protective role against DSA development during the 1st year after KT. Additional inhibitory mechanisms participating earlier in DSA development after KT deserve to be investigated. Summing up, we have presented data that suggests that CD4


Hand hygiene (HH) is a key measure to prevent healthcare-associated infections. Between 2006 and 2008, the WHO conducted pilot testing of the implementation of a multimodal HH improvement strategy in six sites worldwide. Collected data presented several aspects of complexity and levels of clustering that needed to be taken into account in the analysis. We describe a statistical approach aimed at minimizing potential bias arising from such complex datasets.Through a before/after observational study, HH compliance was assessed in several wards ofdifferent hospitals in five countries from April 2006 to December 2008 using a validated method. The HH opportunity was the unit of analysis; data on HH indications and actions, observed professional categories, and day of the week observed were available. Simple logistic regression and mixed models using a nested clustering effect  were used to assess the effect of the WHO strategy.A total of 45 420 HH opportunities were collected during 3613 sessions in 94 wards inin 43 hospitals from six pilot sites. Using a logistic regression model with fixed effects only, the crude odds for HH compliance after the intervention were 1.71 (95% CI: 1.65-1.78) and the adjusted odds, 1.80 (95% CI: 1.73-1.88). With a mixed model, the crude odds for HH compliance after intervention were 1.99 (95% CI: 1.86-2.14) and 2.13 (95% CI: 1.97-2.29) after adjustment.The effect of the strategy implementation may be underestimated if the correlation existing between data is not taken into account. A generalized linear mixed model using a nested clustering effect may be a solution to deal with correlated data, but as paired data are lacking, all correlations cannot be taken into account.None declared.


In this study, we attempted to assess whether the early plasma lactate (PL) level is a useful biomarker to predict septic complications and outcome in burn patients.A retrospective study was conducted in the burn care center in Tunis. Patients admitted within 24 hours from the thermal injury, from 1 January 2009 to 30 June 2010, were included. PL was measured early in the first 24 hours and controlled more than twice. For each measurement, 5 ml venous blood was drawn into a heparin-coated syringe. The normal lactate value was defined as 1 ± 0.5 mmol/l.Over an 18-month period of study, 80 patients were enrolled. There were 60 males and 20 females. The mean age was 40.7 ± 19.5 and the average TBSA was 32 ± 21%. Upon admission, patients with an initial lactate value of more than 2 mmol/l were 86.7%. Fifty-eight percent of them have a lactate initial value of more than 4 mmol/l. In order to evaluate the potential impact of using early lactate measurements (H24 post burn injury) as predictor biomarker of sepsis in burn patients, a linear discrimination function was performed, by measuring the area under the ROC curve, and found that initial lactate value of more than 4 mmol/l provides the best sensitivity and specificity: 88% and 79% respectively. Also, the PL cut-off value for prediction of mortality was 4 mmol/l with a good sensitivity (86%) and specificity (92%). The area under the ROC curve was 0.96.Lactate appears to be a powerful predictor biomarker of sepsis and mortality in burn patients. A serum lactate of 4 mmol/l provides the best sensitivity and specificity.