FEV1, FVC and MEF25%–75% was measured before, 4 h and 24 h after exposure. Wilcoxon-rank sum tests (comparing exposure scenarios) and mixed linear regression using particle concentrations and adjusting for personal characteristics, travel time and transportation means before exposure sessions were performed. While no effect was seen comparing the exposure scenarios and in the unadjusted model, inverse associations were found for PMC from CB and FS in relation to FEV1 and MEF25%–75%. with a change in 10 µg/m3 in PM2.5 from CB being associated with a change in FEV1 of −19 mL  after 4 h. PMC from TB and PNC of UFP were not associated with lung function changes, but PSC from CB was. Elevated indoor fine particles from certain sources may be associated with small decreases in lung function in healthy adults.Particulate air pollution is linked to impaired respiratory health. We analyzed particle emissions from common indoor sources (candles burning (CB), toasting bread (TB), frying sausages (FS)) and lung function in 55 healthy volunteers (mean age 33.0 years) in a randomized cross-over controlled exposure study. Lung-deposited particle surface area concentration (PSC), size-specific particle number concentration (PNC) up to 10 µm, and particle mass concentration (PMC) of PM Exposure to ambient particulate matter (PM) is linked to increased morbidity and mortality with over a million premature deaths worldwide ,2,3. TheIncreased levels of ambient PM air pollution have been associated with asthma exacerbations, increased respiratory symptoms, decreased lung function, and increased hospital admissions for respiratory symptoms due to asthma, chronic obstructive pulmonary disease, and other respiratory ailments as well as for cardiovascular disease ,6,7.Ultrafine particles (UFP) defined as particles with aerodynamic diameters below 100 nm received special attention recently  related et al. examined 13 different indoor particle sources in a full-scale chamber [To date the majority of studies investigated ambient particles, although in most industrialized countries people spend most of their time indoors  and sign chamber . They foSeveral controlled exposure studies so far investigated the effects of diesel engine exhaust, wood smoke and strand board emissions on the respiratory and cardiovascular system ,21,22,23i.e., size-specific particle mass concentration (PMC) with novel exposure metrics such as size-specific particle number concentration (PNC) and lung deposited surface area concentration (PSC) to detect a link between special characteristics of the emitted particles and lung function.In a controlled chamber exposure study, we investigate whether exposure to particles from candles burning, frying sausages and toasting bread as common indoor activities leads to changes in lung function in healthy volunteers. We perform an in-depth characterization of emitted particles combining established exposure metrics The Effects of ultrafine Particles from Indoor Activities (EPIA) Study is a cross-over sham-controlled exposure study with healthy volunteers, integrating a detailed exposure characterization, the investigation of biological pathways with toxicological methods, and human health effect analyses. The study was performed with approval of the Ethics Committee of the Heinrich-Heine University of Düsseldorf, in accordance with the declaration of Helsinki, and with written informed consent of all participants.etc.), fatal disease in the end stage, heart attack or stroke in the past 3 months, uncontrolled arterial hypertension without medication, diabetes mellitus, chronic respiratory disease, chronic infectious disease and neurologic or mental disease .Out of a convenience sample of 154 subjects who were interested in the study, a total number of 55 adult men and women met inclusion criteria and could be recruited for this study. Inclusion criteria comprised age between 18 and 79 years, speaking and understanding German, and being a non-smoker or ex-smoker for at least ten years. Predefined exclusion criteria were: existing or planned pregnancy (within the next six months), planned surgery in the following six months, occupational exposure to environmental tobacco smoke or air pollutants . Flow rate was approximately 250 m3/h. The air conditioning system worked in a circulating mode, hence not causing additional air exchange. Particle monitors operated during the exposure periods at a time resolution of minutes showed a relatively constant steady-state particle concentration, any fluctuation of concentrations was included in the exposure values. The sampling ports were installed close to the seats where the participants remained during exposure, distances ranged between 0.5 and 1.5 m. The vents (air conditioner and active air changer) ensured a good dispersion within the chamber. Tests made by handheld monitors  at various locations around the participant’s seating area revealed concentrations comparable within the instrument uncertainties.Each exposure session lasted for two hours and took place in an air-conditioned laboratory room as exposure chamber. The chamber had an area and a volume of approximately 16 mParticipants were exposed on separate occasions at the same time of day at least two weeks apart. Participants were not blinded against the type of exposure because the particle sources were placed inside the exposure chamber so that the subjects could see and smell them. For the sham exposure with room air (RA), an air ventilator, was placed in the chamber. Participants did not exercise during exposure sessions so breathing patterns correspond to usual quiet indoor activities.®-coated pans for 10 min without additional fat. To avoid extensive accumulation and burning of residues the pans were cooled down and cleaned after three cycles of frying. For this, the active emission was stopped transiently for about 10 min after each 0.5 h frying period.In the experimental exposure scenarios, participants were exposed to candle burning (CB), toasting bread (TB) and frying sausages (FS). Indoor sources were operated by staff being present continuously inside the chamber. Exposure scenarios were performed on two different exposure levels for each emission source. During CB, 20 white Christmas-tree candles  or 40 candles  were burning simultaneously. During the exposure scenario TB, bread was toasted with one  or two  identical time controlled 2-slice-toasters. The duration of each toasting period lasted up to 3 min. To ensure constant emission and in order to prevent the toaster from overheating, the toasting process was done alternately in different toasters of the same type. During the exposure scenario FS, three sausages per pan were fried simultaneously in one  or two  Teflon2/cm3, using a Nanoparticle Surface Area Monitor . Additionally size-specific mass concentrations of PM1, PM2.5 and PM10 were calculated from particle size and number concentrations assuming spherical particles and a particle density of 1 g/cm3. During exposure, temperature and relative humidity was controlled and temperature maintained at 24 °C by a re-circulating air conditioning system installed in the chamber’s ceiling.Particle concentration in the exposure chamber was monitored continuously during each session to ensure consistent particle pollution levels for all participants and each exposure scenario and to calculate the individual cumulative particle exposure for each participant. Before a subject entered the exposure chamber the sources had already been activated to ensure that the particle concentration had reached the required equilibrium level. Size-specific number concentration of particles from 5.6 nm to 562 nm size range was monitored by a Fast Mobility Particle Sizer  and for particles from 0.5 to >10 µm with an Aerodynamic Particle Sizer . Further, the alveolar deposited surface area particle concentration was measured in µm1) and averaged forced expiratory flow between expiration of 25% and 75% of total FVC (MEF25%–75%) were assessed before, and 4 h as well as 24 h after exposure using the ndd Easy One-world-spirometer  with provided software . Spirometry was performed according to the American Thoracic Society/European Respiratory Society guidelines [1 was reached, were used [Forced vital capacity (FVC), forced expiratory volume at one second (FEVt1 = 4 h and t2 = 24 h) were quantified as the difference to pre-exposure (t = 0) values. To estimate exposure related changes in lung function (ytik) at each time point t computed for different indoor sources  and exposure levels  in contrast to room air (RA) (i = 0), the difference-in-differences  was calculated and tested using the paired Wilcoxon Rank-Sum test:Statistical Analyses were performed using SAS version 9.2  and R . Exposure and spirometry measurements are presented as a mean ± standard deviation. Changes in lung function post exposure , PNC (for UFP), and PSC. The effects for the two different time points were modeled with an interaction term incorporating an indicator variable for time point. In addition to the crude model, we adjusted for age, height, sex, temperature, humidity, travel time and means of transportation (full model). The models were fit via linear mixed models from the R language package.Additionally, in a multivariate analysis, we used the personal cumulative exposure to the particle metrics size-specific particle mass, particle number and surface area during the exposure sessions as independent variables and the intra-individual difference to 3) in level 2. Size-specific (<100 nm) PNC at level 2 was approximately twice as high as that for level 1 for all exposure scenarios. Contrary to that, PNC of PM1, PM2.5 and PM10 for CB and TB in level 1 was two to four times higher than in level 2, but not for FS. The mass-concentration for PM1, PM2.5, and PM10 at level 2 was on average at least one and a half time larger compared to level 1, except for PM10 at TB, where the PMC at level 2 was found to be smaller than at level 1. In general, the mass concentration was dominated by the contribution of ultrafine particles, which were a factor 104 to 105 higher in number than particles with size >500 nm. Hence, calculated PMC values in most cases follow the pattern of PNC for UFP. TB was an exception where a considerable decrease in large particle number concentration from level 1 (approx. 20 cm−3) to level 2 (approx. 4 cm−3) caused a decrease in PM10 PMC accompanied by a rising UFP number concentration. This is driven by one particular TB exposure session that was extreme in terms of the number of particles larger than 0.5 µm leading to high PM1, PM2.5 and PM10 exposure. Excluding the three participants of this exposure session reduces the average PM concentrations of TB level 1 down approximately half of the level 2 value.The study population shown in The highest mass concentrations were observed during FS with the maximum at level 2. The surface-concentration was also roughly twice as high as at level 2 compared to level 1 for CB, TB and FS and was found to be nearly the same for all exposure scenarios at level 2.1 decreased 4 h and 24 h after the exposure to RA and FS at level 1 and level 2, whereas it increased 4 h and 24 h after CB at level 1 and TB at level 2. A similar non-uniform pattern is apparent for FVC. Regarding FEV1/FVC, we observed mostly an increase after 4 h and 24 h after the particle exposure while the expiratory flow at 25%–75% of the vital capacity (MEF25%–75%) shows a more heterogeneous picture with mainly increases but a stable value at RA  to −23  for CB and from 2  to −11  for FS when adding “travel time” and “means of transportation” to the model containing already the other covariates ) showed a trend towards a decrease with MEF25%–75%, only. This seems to be in contrast to prior toxicological findings, showing stronger associations for UFP than for fine particles of the same chemical composition and on an equal mass basis [3, comparisons between particle metrics cannot be made directly and have to be viewed with caution.The observed associations of particle metrics on lung function were visible more often for size-specific particle mass in contrast to novel metrics such as particle surface area or size-specific particle number concentration. Associations were mostly observed for PMss basis . However2), and the intra-individual changes in means of transportation , travel time and therefore ambient air pollution exposure before the start of the controlled exposure. Nevertheless, one should be cautious given that the confidence intervals generally increased considerably after adjustment, concomitant with an important change of the effect estimates.The results of the full regression model, adjusted for personal characteristics as well as for time in travel and means of transportation on the morning before exposure, showed some consistent short-term decreases in lung function for candle burning and frying sausages. In contrast, the crude models did not exhibit these effects. Similarly, the analysis of difference-in-differences, directly comparing changes to pre-exposure values for each exposure scenario with changes after room air exposure, had no consistent effect on any lung function variable. This is may be due to better resolution of exposure with individual-specific PM-concentrations and points to the potential importance of taking the individual (pre-)exposure to traffic-related air pollution prior to the controlled exposure into account, especially given the large catchment area of our participants  leads to a temporary decline in PEF in healthy subjects, along with self-reported irritations in upper airways [1 and FVC, may be less sensitive for picking up small effects of short-term exposures in healthy subjects and more sensitive indices or instruments might be necessary.Our results show most consistent associations for MEFollution ,21,22,23 and FVC ,18,19. Aobserved , similarBiological pathways for the observed effects may lead to constriction due to either include airway inflammation or neurogenic responses. Depending on the size of inhaled particles, they deposit in the nasopharynx, enter the main bronchi or even reach the alveoli. PM inhalation can cause inflammation and oxidative stress response within the lungs, detectable through an increase of e.g., cytokines and white blood cells. The inflammatory response will lead to a swelling of the bronchial mucosa, decreasing the overall diameter of the small airways. Furthermore, it can lead to an activation of bronchial smooth muscle cells, causing a subclinical constriction of the small airways . Chemicaet al., reviewed panel studies on pulmonary effects of people living in areas where the exposure was dominated by residential wood combusting, summarizing that the most information was only found for acute asthma in relation to particulate matter with an aerodynamic diameter of <10 μm [et al. [et al. [3 PM10 [1 (34 mL) and FVC (37 mL) per increase of 20 µg/m3 PM2.5 [25%–75%, as well as decreases in FEV1, and FVC in relation to ultrafine particles, and less consistently for PM2.5, which clearly reduced only MEF25%–75%. In contrast to these studies on presumably more susceptible asthmatic patients, we studied changes in healthy participants. Even though the participants in our study were thought to be less susceptible than asthmatic patients, we found effect estimates, that although they are relatively instable, compare well in size with those found in asthmatics [Several panel or controlled exposure studies investigated asthmatic participants, as these are believed to be especially susceptible to the effects of inhaled fine and ultrafine particles. Strong evidence for short-term effects on lung function has come from panel studies in asthmatic children ,28,29,30f <10 μm . The res [et al.  showed tm3 PM2.5 , in contm3 PM2.5 . The autthmatics ,27,30,31et al., the highest particle number concentrations were reached during candles burning at level 2 and were dominated by UFPs [2.5 and PM10, possibly due to fatty droplets which form larger particles.In our exposure assessment the concentration levels  at CB, TB and FS differed strongly to the sham exposure. For size-specific PMC, exposures were 10 to 50 times higher and for size-specific PNC they were 100 to 900 times higher than during room air exposure. For all exposure metrics and for all exposure session we could achieve a sufficient contrast between level 1 and level 2, allowing exposure-dependent analyses. In each of our exposure scenarios we observed very high number concentrations of UFPs. Similar to Afshari  by UFPs . InteresDespite the consistently high concentration levels during all exposure scenarios, observed associations with lung function seemed to differ between exposures, which may reflect different source-specific constituents and resulting differential toxicity of the emitted particles. Toxicological investigations have demonstrated that, depending on their chemical composition and source, the toxic potency of outdoor PM samples of similar size fractions can vary considerably ,33,34.10 and PM2.5 resulting from candles burning and frying sausages were 2 to 12 times higher during chamber exposure than ambient daily mean concentrations, measured at an urban background location in the highly industrialized and urbanized Ruhr area in Germany [et al., 2003, found out that mean indoor PM2.5 over a 14 days period was 31 μg/m3 in homes with cooking and 23.5 μg/m3 in homes without cooking, with an average increase of about 3.5 μg/m3 due to the average number of frying events (nine per week) [vice versa) should generally not be undertaken.In our study we specifically investigated effects of indoor sources of fine and ultrafine particles. Even though a large part of particles found indoors originate from outdoors , indoor sources can potentially result in relevant personal cumulative exposures in rooms with insufficient ventilation. Particle mass concentrations for PM Germany  and comper week) . Cumulatx). We have not measured O3 that could also potentially alter lung function. It cannot be excluded that the exposures under real-life conditions could provoke different effects, all the more as exposure levels may be considerably lower than those that were chosen for the present study. Regardless of the type of exposure, the exposure time of two hours was short and possibly prevented us from observing more clear and consistent effects on lung function variables. The selected time points for the examination of lung function have been selected on the basis of previous studies who also measured three to 6 h and 22 to 24 h after exposure, respectively [Our study has several limitations. Associations could be observed only in the multivariate regression analysis but not in the unadjusted results of the analysis of difference-of-differences between exposure scenarios. While the adjustment for relevant covariates helps to address potential confounding, confidence intervals increased considerably. Furthermore, the advantage of investigating several different exposure metrics is counterbalanced by the possibility to have chance findings due to multiple comparisons. Indeed, given the high number of comparisons, our results could, in the worst case, be the result of chance findings. This is a clear drawback of our analyses which reflects therefore the results of an exploratory analysis and should by no means be interpreted in terms of conclusive statistical tests. Due to sick leave, premature study discontinuation or other personal reasons not all participants received all exposure scenarios. To create controlled and standardized exposure scenarios which are as uniform as possible across study participants, we were not able to eliminate differences to real-life exposures in a common household. The candles for example have been replaced before burning down, decreasing soot emissions and the sausages were fried in a pan without fat. One major limitation is that during the exposure of the participants we did not measure gases that were emitted at the same time and are often irritants and therefore could have affected lung function. Spurious results could result if these gases correlated strongly with emitted particle concentrations. However, measurements during the exposure characterization with a proton-transfer-reaction mass spectrometer  showed sub-ppm levels for all detectable gas-phase compounds (e.g., aldehydes, alcohols, organic acids, NOectively ,19,23,31The strength of our study is a detailed exposure characterization, including continuous measurements of number size distribution, size-specific mass concentrations and surface concentrations, enabling us to assess personal cumulative exposures for established and novel particle metrics. This improves exposure assessment over the mere classification as CB, TB and FS that was used for the test. We achieved a high exposure contrast between the sham exposure and the experimental exposures for all particle metrics, which increased our power to detect even small, clinically not relevant effects.In our study of short-term exposure of healthy adults, we found some associations of fine particles emitted from common indoor sources in the fully adjusted model that also took account of the means of transportation on the day of the exam. However, no consistent effect was seen comparing merely the exposure scenarios or in the unadjusted or partially adjusted model, In the fully adjusted model, associations of fine particles emitted from candle burning and frying sausages with small negative changes in lung function variables were observed. Toasting bread showed no association with lung function. The novel particle metric size-specific PNC was not associated with reduced lung function and PSC showed associations only for candles burning. Common household activities such as candles burning, toasting bread and frying sausages may cause particle mass concentrations indoors, which can possibly cause transient negative changes in respiratory function. Thus, elevated indoor fine particles from certain sources may be associated with small decreases in lung function in healthy adults. However, because we cannot exclude the possibility of our results being in line with chance findings from multiple comparisons, further in depth investigations are necessary to corroborate these results and to investigate potential biological pathways involving local pulmonary inflammation and more sensitive markers of small airway constriction.


Chromium (Cr) is commonly used as a complementary medicine for diabetes mellitus. Several studies suggest that Cr intakes may improve glucose metabolism and decrease oxidative stress. Therefore, we aimed to assess the effects of chromium histidinate (CrHis) supplementation using a range of reliable biomarkers of oxidative damage and histopathological changes in rats with diabetic retinopathy.Diabetes was induced with streptozotocin  by intraperitoneal injection in male Long-Evans rats. Three weeks after STZ injection, rats were divided into four groups, namely, untreated normal controls, normal rats receiving CrHis (110 μg/kg/day); untreated diabetics and diabetics treated with CrHis (110 μg/kg/day) orally for 12 weeks.In the untreated diabetic group, levels of serum glucose, glycosylated haemoglobin (HbA1c), total cholesterol (TC) and retina malondialdehyde (MDA) were significantly increased, while expressions of retina insulin, and glucose transporter 1 (GLUT 1) and glucose transporter 3 (GLUT3) and level of serum insulin were decreased. CrHis supplementation was found to reduce the levels of glucose, HbA1c, total cholesterol and MDA and to improve the GLUT1, GLUT3 and insulin expressions in STZ-induced diabetic rats. CrHis prevents the changes in the expressions of GLUT1, GLUT3 and insulin and the level of MDA in the retina tissue, confirming the protective effect of CrHis supplementation against the retinopathy caused by STZ. Histopathologic findings suggest that the CrHis-treated diabetic group had normal retinal tissue appearance compared with the untreated diabetic group.These results verify that CrHis has critical beneficial effects against retinal complications. Although detailed studies are required for the evaluation of the exact mechanism of the ameliorative effects of CrHis against diabetic complications, these preliminary experimental findings demonstrate that CrHis exhibits antidiabetic effects in a rat model of diabetic retinopathy by regulating the glucose metabolism and suppressing retinal tissue damage. Diabetic retinopathy is the most common diabetic eye disease and a leading cause of blindness in working-age population. The pathophysiology of diabetic retinopathy is complex and multifactorial. The pathogenic process involves intricate interactions between oxidative stress and hyperglycemia ,2. OxidaIn a high energy-demanding tissue such as retina, the regulation of glucose uptake and its utilization is important for the maintenance of normal retinal function. It is well known that glucose uptake is regulated by glucose transporter proteins (GLUTs) and all mammalian cells contain one or more members of this GLUT protein family. Among these transporters the GLUT3 and GLUT1 isoforms have been expressed in retinal cells ,8. GlucoThe incidence of diabetes mellitus is associated with various events such as obesity, increasing age, physical inactivity etc. Furthermore, certain nutritional deficiencies have also been associated with a higher incidence of diabetes mellitus ,9. ChromTwenty eight Long Evans rats, useful animal model for studying ocular diabetic complications, aged 8 weeks with 250 ± 20 g of body weight were obtained from Ertugrul Kilic . All the animals were kept and maintained under the standard guidelines and housed in the Veterinary Control Institute . The animals were kept and maintained at 22 ± 2°C, humidity of 55% ± 5% and 12/12-hour light/dark cycle. The rats were allowed free access to feed [standard pellet diet] and drinking water. The experiment was approved by the Animal Care and Use Committee, University of Firat with ethics number: 2011-10-131. All procedures involving rats were conducted in strict compliance with relevant laws, the Animal Welfare Act, Public Health Services Policy, and guidelines established by the Institutional Animal Care and Use Committee of the University. The rats were weighed every week and at the end of the study. Blood sample was collected from the tail vein of each rat for the measurement of biochemical efficacy and safety markers.The CrHis  was given in the water and administered at a concentration of 110 μg/kg bw/d) to get 9.16 μg elemental Cr (kg body/d), which is an equivalent dose of 614 μg Cr for a 70-kg adult human based on a previous work . A P value of less than 0.05 was considered significant.P < 0.5). When diabetic retinopathy rats were treated with CrHis, significant increases in blood glucose, HbA1c, insulin and total cholesterol levels were observed in diabetic retinopathy rats. CrHis treatment also resulted in a significant decrease in mean serum total cholesterol concentration of diabetic retinopathy animals. Body weight was significantly decreased (P < 0.001) in the untreated diabetic rats when compared to control group. CrHis treatment significantly increased body weight (P < 0.001) compared to the untreated diabetic group . Blood glucose, HbA1c and total cholesterol levels were significantly increased in untreated diabetic rats compared to control groups while insulin levels were decreased  in the retina of diabetic rats compared to control. Treatment using CrHis significantly (P < 0.05) reversed these changes to near control levels . A statistically significant reduction of MDA expression was found in retina of diabetic rats when the diabetic rats were treated with CrHis  rats, with intact layers. The retinas were disorganized in the diabetic rats with impaired layers. But the retinas in CrHis group were improved compared to the diabetes group Figure .Figure 2Diabetic retinopathy is the most common microvascular complication of diabetes. It remains a major cause of vision impairment or blindness in the world and high blood sugar level is considered the chief instigator in its development. The pathogenetic mechanisms of diabetic retinopathy are rather obscure. Many hyperglycemia-induced metabolic abnormalities are implicated in its pathogenesis, such as increased oxidative stress, and increased advanced glycation endproduct formation and other tissue proteins, which are observed during long-term hyperglycemia . The retThe risk of loss in visual acuity was correlated with the degree of retinal hard exudates, reducing serum lipid levels in patients with diabetic retinopathy . The mosAs one of the essential trace elements, trivalent cr plays a significant role in the regulation of glucose and insulin metabolism. Chromium is biologically active as part of an oligopeptide–chromodulin (low-molecular-weight chromium binding substance)–potentiating the effect of insulin by facilitating insulin binding to receptors at the cell surface. With Cr acting as a cofactor of insulin, Cr activity in the organism is parallel to insulin functions . Cefalu Chromium has consistently indicated improvement in glucose transport in various tissues both in vitro and in animal models. Several mechanistic studies have demonstrated improvements in insulin receptor/postreceptor signaling, leading to enhanced activity of GLUTs. Interestingly, Cr can directly bind insulin, making the insulin more stable and possibly prolong its duration of action. Chromium treatment also helps the translocation of GLUTs and increases membrane fluidity by reducing the plasma membrane cholesterol . The retIn conclusion, the results of the present study demonstrate the benefits of supplementation with CrHis in rats with diabetic retinopathy. CrHis supplementation offer improvements to markers of disease risk, including hyperglycemia insulin sensitivity, and increased oxidative stress. These findings are encouraging and warrant further clinical trials of this promising CrHis supplementation.


Therefore, the aim of this study was to assess if bacterial DNA from primarily oral bacteria could be detected on coronary angioplasty balloons by use of an optimized sampling process combined with an internally validated sensitive polymerase chain reaction (PCR) assay. Coronary angioplasty balloons and control samples from a total of 45 unselected patients with stable angina, unstable angina/non-ST elevation myocardial infarction, and ST-elevation myocardial infarction (n = 15 in each group) were collected and analyzed using a PCR assay with high sensitivity and specificity for 16S rRNA genes of the oral microbiome. Despite elimination of extraction and purification steps, and demonstration of sensitivity levels of 25–125 colony forming units (CFU), we did not detect bacterial DNA from any of the coronary angioplasty balloons. A subsequent questionnaire indicated that the prevalence of periodontitis in the study cohort was at least 39.5%. Although coronary angioplasty balloons are unlikely to be useful for detection of bacteria with current PCR techniques in unselected patients with coronary artery disease, more studies are warranted to determine the extent to which bacteria contribute to atherosclerosis and its clinical manifestations and whether the presence of bacteria in the arteries is a transient phenomenon.Periodontitis is a chronic, bacterially-induced inflammatory disease of the tooth-supporting tissues, which may result in transient bacteremia and a systemic inflammatory response. Periodontitis is associated with coronary artery disease independently of established cardiovascular risk factors, and translocation of bacteria from the oral cavity to the coronary arteries may play a role in the development of coronary artery disease. Very few studies have used angioplasty balloons for  Chlamydophilia pneumonia, Helicobacter pylori, Porphyromonas gingivalis and Cytomegalovirus (CMV) [Cardiovascular disease (CVD) is the leading cause of death worldwide and its primary cause, atherosclerosis, is a chronic inflammatory disease of the arteries, which is strongly associated with risk factors such as hypertension, smoking, diabetes, obesity, hyperlipidemia, a family history of CVD, and socioeconomic deprivation. However, for more than three decades it has been hypothesized that infection may play a role in the atherosclerotic process from initial endothelial dysfunction to clinical manifestations, e.g., myocardial infarction (MI) and stroke –8. Previus (CMV) ,9–11. Hous (CMV) –8,12.Periodontitis, occurring in as much as 50% of the population over 50 years of age, is a polymicrobially-induced chronic inflammatory disease in which multiple bacterial species, including anaerobic Gram negative rods and spirochetes organized in a biofilm, colonize the periodontal pockets . The resin vivo by use of angioplasty balloons from unselected patients undergoing PCI, 2) the severity of the clinical manifestations of coronary atherosclerosis, i.e., stable angina, unstable angina (UAP), non ST-segment elevation MI (NSTEMI), or ST-segment elevation MI (STEMI) in these patients was proportional to the amount and diversity of the bacteria detected on their respective angioplasty balloons.The usefulness of coronary angioplasty balloons for retrieval of atherosclerotic tissue material for detection of bacteria has previously received very limited interest and results have been both positive and negative –37. Duride novo coronary lesion were enrolled at two tertiary invasive cardiology centers , including 15 patients with stable angina, 15 patients with UAP or NSTEMI, and 15 patients with STEMI. Patients with a history of infection requiring systemic antibiotic treatment and those with immunocompromising conditions, e.g., chemotherapy or chronic inflammatory diseases receiving immunosuppressive treatment, within a three months period prior to admission were excluded from the study as were subjects with clinical and biochemical signs of active infection. After a mean 17 months after PCI, two patients had died and the remaining patients were contacted by phone and subjected to a standardized questionnaire to determine if at the time of PCI they had periodontitis diagnosed by a dentist, the number of remaining teeth, the number of loose teeth, and whether gum bleeding occurred regularly when performing oral hygiene. The study was approved by The Regional Committee on Health Research Ethics of The Capital Region of Denmark (protocol number H-2-2012-154) and written, informed consent was given by all participating patients.A total of 45 patients subjected to PCI of a Immediately upon withdrawal of the angioplasty catheter from the guiding catheter, the distal balloon part of the catheter (approximately 20 mm in length) was cut directly into sterile, RNAse/DNAase-free tubes, sealed, labeled and quickly frozen at -80°C. Only angioplasty balloons used for predilation were sampled and subsequent balloons used for drug-eluting stent implantation in individual patients were specifically avoided, due to their polymeric coatings and cytostatic drug release which could potentially interfere with bacteria and the PCR assay. As negative controls we used a 20 mm control segment of the respective angioplasty catheter taken 5–7 cm proximally from the balloon. This part of the catheter was in contact with the arterial blood and potentially  in passive contact with the arterial wall of the proximal coronary artery segments. If the electro-chemical properties of angioplasty catheter material promoted passive adherence of circulating bacteria from the blood  these catheter control segments were expected to be positive to the same extent as the angioplasty balloons. In addition, to control for positive samples being the result of contamination from arterial blood, blood samples were drawn from the femoral artery sheath and transferred into heparinized collection tubes. All sample collections were done using single use sterile equipment and samples and controls were stored at -80° C until analysis.Escherichia coli as reference). This specific choice of primer pair was based on matching the sequences of several combinations of commonly used ‘universal’ 16S RNA primers with the online 16S RNA library of the Human Oral Microbiota Database (HOMD) [C. pneumoniae, H. pylori and P. gingivalis. For each analysis the forward primer sequence was fitted at the 5’-end with a 42–43 bp barcode sequence with unique recognition sequences allowing downstream deep sequencing of PCR products by 454 pyrosequencing. The reverse primer was tagged with a non-unique 31 bp sequence. Method optimization and validation was done using barcoded and tagged primers to ensure that the added sequences did not interfere with the specificity and sensitivity of the assay. The master mix contained 50 μL 2x Phusion Blood PCR Buffer  containing dNTPs, 38.5 μL DNAse/RNase and proteinase free molecular water , 0.25 μM primer 357F-barcoded and 0.25 μM primer 1391R-tagged, 2 μL (2%) DMSO and 2 μL Phusion Blood II DNA polymerase . Total reaction volume was 100 μL, assuming a 5 μL volume for template in the form of angioplasty balloon, catheter control segment or 5 μL arterial blood. Amplification was performed on a Applied Biosystems VeritiTM 96-Well Thermal Cycler  with 10 min initial denaturing at 98° C following 45 cycles of denaturation at 98° C for 5 sec, annealing at 63° C for 5 sec and extension at 72° C for 15 sec. Final extension was at 72° C for 1 min. Solid material in the PCR tube was pelleted by centrifugation at 1000g for 5 min and 10 μL of the supernatant was run on a 1% EDTA agarose gel for 30 min at 90V. The DNA ladder used was Thermo Scientific GeneRuler 1kb DNA Ladder . A positive signal was defined as a single visible band on the gel of the anticipated length.The collected samples were thawed on ice in random order, and the angioplasty balloons and catheter control segments were cut in smaller pieces, allowing them to fit directly into a pre-prepared PCR master mix in standard 200 μL PCR strips. Five microliter of the arterial blood sample was likewise pipetted directly into prepared PCR tubes. Measures to minimize risk of contamination during analysis were carefully undertaken, and all handling was done in laminar flow cabinet using sterile equipment. PCR master mix, flow cabinet, and equipment were UV-radiated for 10–20 min prior to use, to reduce risk of DNA contamination. PCR was done using Thermo Scientific Phusion Blood Direct PCR Kit . To allow for identification of a broad spectrum of bacteria we used primers annealing to the highly conserved regions of the bacterial 16S rRNA gene. The forward primer was 357F (5’-TCC TAC GGG AGG CAG C-3’) and reverse primer was 1391R (5’-GAC GGG CGG TGT GTR CA-3’) generating a 1034 bp product (using the 16S rRNA gene of e (HOMD) . This waPseudomonas aeruginosa (PAO1) as model for Gram negative bacteria and Streptococcus mutans (UA159) as model for Gram positive bacteria. Dilution rows of blood with bacteria were prepared in 1.5 mL Eppendorf tubes. Sterile angioplasty balloons  were dipped into each Eppendorf tube and dilated for 5–10 sec, then deflated and cut into sterile RNAse/DNAas free tubes and frozen at -80° C for 24 h before analyses using the method described above. Repeated weighing of angioplasty balloons, before and after they were dipped in blood, revealed that the amount of blood able to adhere to the surface and in the folds of deflated angioplasty balloons varied between 1 and 5 μL. Therefore, 5 μL of arterial blood from each tube of the dilution rows were used as control for the angioplasty balloons. The bacterial concentration of the dilution rows was determined by spectrophotometry validated by cultivation and use of microscopic counting chambers.Validation of the PCR assay was based on repeated experiments using dilution rows of human blood from healthy volunteers spiked with either P. aeruginosa), depending on the amount of blood (1–5 μL) adhering to the angioplasty balloon. The detection limit was 125 CFU, when analysis was done directly on 5 μL spiked blood without angioplasty balloon (S. mutans) the detection limit in repeated experiments was somewhat higher (102−103 CFU).Detection limits were between 25–125 colony-forming units (CFU) for angioplasty balloons dipped in blood spiked with a Gram negative bacterium  were positive  were negative, and all positive controls  who underwent PCI. The periodontal status of the patients assessed by our standardized questionnaire suggested a prevalence of periodontitis of at least 39.5% which is comparable to other published data and clearly indicates that the negative results were not explained by absence of periodontal disease .Firmicutes being the most predominantly represented phylum from subgingival samples, and Proteobacteria dominating the findings from coronary angioplasty balloons. Indeed, many of the species that were isolated exclusively from angioplasty balloons are not known to be indigenous to the oral cavity although some species, including P. gingivalis, were isolated from both subgingival samples and coronary angioplasty balloons, which may be indicative of a possible translocation from the oral cavity [Several previous studies have reported on bacterial findings from endarterectomy samples from carotid or coronary arteries, and arteries removed at autopsy, but only very few studies have used coronary angioplasty balloons for sample retrieval and none of these studies reported results according to different manifestations of coronary artery disease . With use of a ‘universal’ 16S RNA primer set, one of these studies reported an apparent 100% detection rate of bacterial DNA from 40 different species retrieved from coronary angioplasty balloons by use of an extraction, amplification, cloning and cultivation method from 18 patients with moderate to severe periodontitis selected from a total of 134 screened patients . These il cavity . HoweverC. pneumoniae on washing medium from angioplasty balloons from 76 unselected patients with de novo coronary artery disease and found traces of this obligate intracellular bacterium in 39.5% of patients [C. pneumoniae and CMV were all negative [Other investigators used a cloning technique and nested PCR with use of selective primers for detection of patients . As contpatients . In potenegative . To the in vivo material from culprit coronary arteries for PCR analysis. Indeed, other researchers have previously examined aspirated intracoronary blood/thrombus from 81 unselected patients with STEMI with use of specific primers for five oral bacterial species and detected the presence of Aggregatibacter actinomycetemcomitans, P. gingivalis, and Treponema denticola in 17, 3, and 2 subjects, respectively [Streptococcus mitis group, and 35 (34.5%) were positive for periodontal bacteria, e.g., A. actinomycetemcomitans, P. gingivalis, and T. denticola [S. mitis and other viridans streptococci [Compared with sample retrieval from coronary angioplasty balloons, catheter-based manual coronary thrombus aspiration during primary PCI in patients with STEMI has more recently provided another method for obtaining ectively . Furtherenticola . Interesptococci . Althougptococci ,46. Impoptococci ,47.In conclusion, with use of an optimized sampling process and internally validated sensitive PCR assay, we were not able to demonstrate the presence of bacterial DNA from the oral microbiome on angioplasty balloons from unselected patients representing the full spectrum of symptomatic coronary artery disease that underwent PCI and where a substantial proportion had a history of periodontitis. Although coronary angioplasty balloons are therefore unlikely to be useful for detection of bacteria with current PCR techniques in unselected patients with coronary artery disease, more studies are warranted to determine whether bacteria contribute to atherosclerosis and its clinical manifestations.


Achieving tight glycemic control for the management of type 1 diabetes is often associated with a heavy burden of care, and frequent hypoglycemia. Closed loop insulin delivery, or an “artificial pancreas”, represents a new technological frontier for the management of type 1 diabetes which aims to overcome these difficulties. Rapid advancement and improvement in the components of closed loop systems  has been translated into numerous published reports demonstrating effective glucose control in hospital studies, camps, hotels, and most recently – in home studies. Most studies to date have focused on overnight control due to the difficulties in managing glucose excursions from carbohydrate intake and exercise. We investigated the capability of the Medtronic Hybrid Closed Loop (HCL) System in managing glucose levels during both day and night. The Medtronic HCL system consists of a Medtronic MiniMed insulin pump, Medtronic MiniMed Enlite II glucose sensor, Medtronic MiniMed Minilink REAL time sensor, Medtronic MiniMed Translator, and an Android mobile device with the HCL algorithm  software application installed. When using the HCL, meal boluses are delivered manually using the Android mobile device. All basal insulin delivery is controlled by the algorithm. We present in-clinic pilot data from 3 individuals with type 1 diabetes, incorporating 140 hours of closed loop management, over 5 days and nights. A free living environment was simulated, with free access to food, and exercise encouraged. For the three participants, percent of time spent in target glucose range (4 – 10mmol/L) was 81%, 62% and 73% respectively. Including all data, the mean glucose was 8.74mmol/L, which corresponds to an HbA1c of approximately 7%. There were no hypoglycemic or adverse events. We conclude that the Medtronic HCL system is potentially effective and safe for the management of type 1 diabetes. Outpatient studies, with comparisons to sensor augmented pump therapy with low glucose suspend in a randomized cross-over trial are ongoing.


Sebastes serranoides), black (Sebastes melanops), and blue rockfish (Sebastes mystinus) are all common inhabitants of nearshore ecosystems on the West coast of North America and important components of the recreational fishery off California. Acoustic monitoring studies indicate that olive rockfish are highly residential and that black rockfish are capable of long migrations and have less site fidelity; yet little is known about the long-term movements of blue rockfish. External tag-recapture studies indicate that blue rockfish may have intermediate movements relative to these congener nearshore species. To better understand the site fidelity, and daily and seasonal movements of blue rockfish over long (>1-year) time scales, we placed acoustic transmitters into 21 adult blue rockfish  in Carmel Bay, California. Blue rockfish displayed intermediate movement patterns and residency relative to other similar kelp forest rockfish species. Two-thirds of tagged blue rockfish (13 fish) exhibited high residency to the study area (>12 mo). When in residence, mean home range of blue rockfish was 0.23 km2, however as many as 30% of tagged blue rockfish shifted their core home range area during the study. Most shifts in home range occurred during upwelling season, and tagged fish moved up to 3.1 km when in residence. Blue rockfish with short residence times were last detected in the study area in late winter and early spring. Blue rockfish were observed at shallower depths during day than night, likely indicative of diurnal feeding. However, over longer time scales, blue rockfish were detected at deeper depths during upwelling periods and with increased wave heights. Daily and seasonal vertical movements of blue rockfish may be influenced by upwelling conditions and local prey abundance.Olive ( Sebastes mystinus), olive (S. serranoides), and black (S. melanops) rockfish are common semi-pelagic nearshore rockfish species in central California that share similar ecological and life history characteristics Blue (2), and utilized even smaller core areas within that home range.Tag-recapture studies, however, lack detailed information about fish movements. With the advent of acoustic telemetry studies, the paradigm of the “nearshore resident” can be more closely evaluated at daily, seasonal, and annual time scales. For example, although tag-recapture data for olive and black rockfish show relatively similar trends in movement, i.e. in most studies, approximately 90% of fish were recaptured <5 km from release 2) for 10 acoustically tagged blue rockfish in northern California, yet the short duration of the study (21 d) and small spatial scale precluded any conclusions regarding seasonal and annual patterns of movements. From these studies, it seems probable that blue rockfish maintain small home ranges for at least part of the year, but whether these home ranges expand or if migration occurs over longer time scales has yet to be determined.Interpreting the long-term movement patterns for the third species, the blue rockfish, is more difficult in the absence of long-term acoustic telemetry studies. Available data on blue rockfish movements are limited to long-term external tagging studies with poor spatial resolution and one high resolution, but short-term (three week) acoustic study. Tag-recapture studies indicate that blue rockfish may have intermediate spatial movements relative to black and olive rockfish. For example, 90% of tagging studies on blue rockfish have reported a mean recapture distance of <3 km, whereas 10% of blue rockfish movement studies have reported larger movements (as great as 41 km) We designed a study to track the movements of adult blue rockfish over a 15-month period using acoustic telemetry in Carmel Bay, California, USA. Our goal was to explore the movement patterns of blue rockfish and determine whether movements of blue rockfish fit the paradigm of the nearshore resident (e.g. olive rockfish), or are characterized by greater seasonal movements that are related to environmental conditions. The specific objectives of this study were to: (1) quantify annual home range and residency of blue rockfish, (2) describe the daily and seasonal movement patterns of blue rockfish with respect to local oceanographic conditions; and (3) estimate factors influencing detection capabilities of acoustic receivers in nearshore physical environments.This study was conducted with the approval of San Jose State University, and carried out in strict accordance with the San Jose State University Animal Welfare Policy according to Academic Senate Policy F06-4. The protocol was approved by the Institutional Animal Care and Use Committee (Permit Number: 824). All surgery was performed under tricaine methanesulfonate (MS 222), and all efforts were made to minimize suffering. This research was conducted under California Department of Fish and Game Scientific Collecting Permit SC-008142.Macrocystis pyrifera, Nereocystis luetkeana and perennial understory algae Pterygophera californica are abundant in the rocky habitats. The majority of the study area is within the boundaries of two Marine Protected Areas (MPAs)  . The Car−1) of MS 222 and Vemco V13P-1H-S256 acoustic transmitters  were surgically implanted into the peritoneal cavity. Transmitters emitted coded signals, which included fish depth, at a random interval between 90–270 s. Predicted battery life for each transmitter was 445 d. An external T-bar anchor tag imprinted with a unique tag identification number also was implanted into the dorsal musculature of the fish in case of angler recapture. Fish were released at the location of capture.We caught blue rockfish with rod and reel fishing gear during September and October 2006. To avoid damage due to barotrauma, we targeted and caught fish in <15 m of water. Large blue rockfish (> length at 50% maturity) We placed 29 acoustic receivers (Vemco VR-2) along the coastline between Pescadero Point and Carmel Point, in depths of 7–40 m . ReceiveAcoustic detection capabilities of VR-2 receivers are affected by biological and anthropogenic noise, sea state, bottom topography, and submerged vegetation The variation in receiver detection ranges over longer time scales was examined by deploying a reference transmitter (Vemco V13-1H-RO4K) 1 m off the seafloor at a fixed location, 140 m equidistant from two receivers located near kelp beds. Reference transmitter data were then correlated with physical parameters including time of day, wave height, mean wave direction, and wind speed. Differences between day and night signal detection for the reference pinger were compared using different filtering methods and tested using a Pearson’s chi-square test. The reference transmitter was deployed from March through September of 2007.http://www.ndbc.noaa.gov). Day lengths were derived from the historical archives of the US Naval Observatory from Carmel, California (http://aa.usno.navy.mil/data/). Daily and monthly upwelling indices, expressed as m3 s–1 100 m–1 of coastline at 122° W, 36° N, were obtained from NOAA Fisheries Pacific Fisheries Environmental Laboratory (PFEL) (www.pfeg.noaa.gov/products/PFEL).Oceanographic and atmospheric data were acquired from the historical data archives of the National Oceanic Atmospheric and Administration (NOAA). Wave height, wind speed, and barometric pressure were recorded from Monterey Buoy 46042, . Occasionally, receivers were temporarily missing from the array because they were lost in storm events. For presence and absence calculations for a given fish, we excluded the dates from analyses for which a receiver was missing due to storms.Diel changes in presence of blue rockfish were characterized by calculating the percentage of hours a fish was detected in the study area during the day versus at night. Day hours were defined as the time period between sunrise and sunset. Night hours were defined as the time from 1 h after sunset until 1 h before sunrise. Crepuscular hours (1 h before sunrise and 1 h after sunset) were excluded from analyses. A fish was considered to be present in an hour if there were ≥2 tag detections in that hour. The number of hours a tagged fish was detected during the day or night on each date was normalized by the total number of night or day hours available on that date and summed to create a contingency table with the expected and observed percentage of hours detected for night periods versus day periods. Fisher’s exact test was used to test the null hypothesis that there was no difference in fish presence in the study area, either between day and night periods annually or for selected seasons.Vertical movements of blue rockfish were examined by comparing differences in depth anomalies of tagged fish. To calculate daily depth anomalies, we subtracted the annual mean depth of a given fish from the daily mean depth of that fish on each date of the study. Depth anomalies were averaged across all fish on each date of the study to obtain a mean anomaly for each date and for day and night periods. Positive anomalies represent deeper mean depths than the annual mean depth; negative anomalies represent shallower mean depths than the annual mean depth.X and Y time series, using these fitted models to obtain estimated residuals, and then determining the cross correlation function between the two residual time series. We conducted the cross correlation analysis for a year period (October 2006–October 2007), the high upwelling season (April–June), and the low upwelling season (December–February), as defined by monthly upwelling indices. To compare mean day depths with mean night depths of fish across the different oceanographic seasons, we used a repeated-measures ANOVA implemented through a mixed-effect model with depth as the dependent variable and fish as the random term. In the model, we tested a time of day effect, a seasonal effect, and an interaction term.We used cross correlation analysis and a generalized linear model (GLM) to compare daily depth anomalies of blue rockfish with water temperatures, the square of mean wave heights, wind speeds, upwelling indices, and atmospheric pressure th percentile of the total hours that a fish was recorded during time at liberty were used to calculate the home range of that fish. In ArcGIS, a polygon with the estimated home range for each individual fish was drawn around these receiver locations, assuming the full receiver detection radius of 150 m from the receiver location. The area encompassing home range for each fish was calculated using the ArcGIS spatial analyst extension, XToolsPro.We calculated a measure of home range for each tagged fish in the following manner. For each hour that a fish was present in the array, it was assigned one or more locations, based on the receiver(s) that recorded signals from the tagged fish. The number of hours detected at each receiver location was tallied for each fish. Receiver locations that ranked in the 90Twenty-one blue rockfish were captured, tagged, and released in the study area between 27 September and 11 October 2006 . FourteePreliminary range trials, conducted before the tagging study began, enabled us to estimate VR-2 detection ranges of 150 m in nearshore areas near kelp forests. Long-term acoustic testing with the stationary reference transmitter, placed 140 m equidistant between two receivers, revealed that the total number of daily detections received was highly variable, ranging from 7–401 detections per day. An average of 9.3±5.9 signals/h was recorded from the reference transmitter during the day and 6.1±4.3 signals/h at night; this difference was significant . When daily  and hourly  detection filters were applied to the data, the reference transmitter was detected for 100% of the days and 91% of the nights it was deployed.2 = 0.5878, p<0.05) was found between the number of hours without recorded signals and Δ T, when the temperature differential between 14 m and 31 m was >1°C.We found no significant (p>0.05) correlations among detections of the reference transmitter and wave height, mean wave direction, or wind speed. Thermal stratification at deeper depths did cause variation in reference transmitter detections. A positive correlation (rResidence times of blue rockfish in the study area ranged from 32.4–100% (144–445 d) of the expected battery life of transmitters . Mean reFive blue rockfish were present in the study area 100% of days during time at liberty after receiver loss was accounted for. Of the remaining 15 fish that did periodically leave the study area, absences were <15% of the total time at liberty. There was no daily pattern of synchronous absences and returns from the study area among fish, although the six fish with the shortest residence times were last detected in the array during the upwelling season (between late April and June 2007). Four of these fish were last detected within the two-week period between 18 April and 2 May 2007, at the peak of the spring upwelling season. Also, absences were recorded for eight fish for 1–3 d during intense storms (4 December 2007 and 5 January 2008), when sea states exceeded 10 m for >24 h.There was no difference in the proportion of hours blue rockfish were detected in the study area between day and night periods , nor for the high and low upwelling seasons . Blue rockfish were detected for an annual mean of 77.7%±3.9 of day hours and 59.5%±5.0 of night hours. The monthly mean percentage of hours in which tagged fish were detected during the day was relatively constant (76–90%) between October 2006 and October 2007. Monthly mean percentage of night hours with detections was consistently lower than the mean percentage of daytime hours (49–68% of night hours). The highest mean nighttime presence occurred in August 2007 (68.3%±7.4).Water depths in the study area ranged from 10–20 m, and tagged blue rockfish were released throughout the study area in depths between 5 and 12 m. Tagged fish often exhibited depth changes of 5–10 m over short time intervals (5–20 min). The mean annual depth of all blue rockfish was 8.7 m±0.6; mean annual depths of individuals ranged from 5.5–12.6 m. An evaluation of blue rockfish depths using a mixed-effects model showed significant interactions between depth and time of day (i.e. day/night) and season (p<0.0001), with no significant interaction between season and time of day . Tagged The GLM analysis indicated significant positive relationships between the depth anomaly and upwelling indices, wave height, and atmospheric pressure (p<0.001). Cross correlation analyses of all data combined indicated that depth anomalies of tagged blue rockfish were positively correlated with upwelling indices with a lag of 1 d (p = 0.001), atmospheric pressure with no lag (p = 0.030) and the square of wave height (p<0.001) with a lag of 1 d . There wWhen evaluated with respect to the months with the highest mean monthly upwelling indices (April–June), depth anomalies were again positively correlated with upwelling indices (p<0.001), atmospheric pressure (p = 0.050), and the square of wave height (p = 0.014), all with lags of 0–2 d . There w2. Mean home range area was 0.23 km2±0.03, and a mean of 2.8±0.3 receiver locations were used in these home range estimations. Regression analysis revealed no significant relationship between total length and home range areas for all fish combined (p = 0.329). Home ranges of blue rockfish were centered near the original site of release for each fish  was obtained from a short-term, small-scale acoustic positioning study Published studies have reported that blue rockfish have small home ranges 2 and 0.25 km2 respectively). Similar to black rockfish One of our hypotheses was that blue rockfish would have a smaller home range than the more mobile, piscivorous black rockfish. However, blue and black rockfish mean home ranges, were nearly equal in Carmel Bay, CA (0.23 km2) Our hypothesis that blue rockfish would have larger home ranges than olive rockfish proved to be true. The mean home range of blue rockfish in our study was several orders of magnitude larger than that reported for olive rockfish (0.0013 kmBlue rockfishes are reliant upon local oceanographic conditions for food and demonstrate immediate feeding responses to increased zooplankton densities in northern California Diel movements of rockfishes have been reported for a variety of rockfish species Similar to previous studies Although blue, olive, and black rockfish occupy the same general spatial areas annually, the vertical distributions of the three species were significantly different from one another during the non-upwelling season Many acoustic studies have been conducted and published without measurement or description of the acoustic underwater environment and how it affects the detection range of receivers. Characterizing the acoustic properties of the water column and how they affect the transmission of animal transmitter signals is critical to prevent false conclusions about movements of tagged animals that may actually be artifacts of the underwater environment. In our study, a reference transmitter was recorded significantly more times during the day than at night. This was likely due to the interference of acoustic signals from biological noise associated with increased nocturnal activity of crustaceans and other marine organisms. We accounted for this observed daily bias in the array's detection capability by combining detections into hour bins; we observed similar transmitter detection rates once the data were filtered and analyzed at the scale of hours. By standardizing the detections of blue rockfish using hours as an indication of presence or absence rather than individual detections, we are confident that differences in detections are due to actual movements of fish and not due to characteristics of the acoustic environment.Blue, black, and olive rockfishes successfully cohabitate in the kelp forest ecosystem but have distinctly different movement patterns. All three rockfishes generally have small home ranges, but different temporal activity and site fidelity to a given location. Olive rockfish maintain high site fidelity and small home ranges year round, and are diurnally active. Black rockfish are nocturnally active, have small home ranges when in residence, and demonstrate the capacity for long-distance migrations


Soil bacterial community structures in terraced rice fields and abandoned lands in a hilly and mountainous area were analyzed using 16S rRNA gene sequences. The DGGE band patterns of each soil were similar. Based on pyrosequencing data, the richness and diversity of bacterial species were slightly higher in paddy fields than in other soils. A beta-diversity analysis clearly indicated that the bacterial community structure in paddy fields differed from those in non-paddy field lands and crop fields that had not been used as a paddy field. These results may reflect the history of land use. Soils are regarded as dynamic living substances and the most diverse microbial habitats on Earth. Bacterial communities in soil are very important particularly for agriculture and vary from soil to soil depending on the environment. Several factors are known to affect bacterial community structures, such as soil physicochemical properties , 13, 15,Agriculture in hilly and mountainous areas is very important in Kochi Prefecture, Japan, because more than 80% of this prefecture is covered by forest and plain land is limited. Although agriculture was widely practiced in mountainous terraced paddy fields more than 30 years ago, many paddy fields have since been abandoned and have remained unused for a long time. A large number of studies have been conducted on the isolation of soil microbes and evaluation of microbial communities for agricultural aspects in Japan , 16. HowCitrus junos, C), planted forests (PF), vegetable crop fields (CF), bamboo thickets , and a ginkgo garden (Ginkgo biloba. G). Soil samples were collected from depths of 0–15 cm and 15–30 cm in triplicate, and were mixed well to make one composite sample that was air-dried and passed through a 2-mm mesh sieve for a general physicochemical analysis. Only soils from 0–15 cm were used in the DNA analysis. Na+, K+, Mg2+, and Ca2+ concentrations were determined using an ion analyzer . Available P was measured by Troug’s method  on March 29 and 30, 2013. Soil samples were collected from seven land use types: paddy fields (P), abandoned paddy fields (AL), Yuzu citrus gardens , 2 vegetable crop fields (CF1 and CF2), a ginkgo garden (G1), and 3 bamboo thickets (B1 to B3). The sites with similar altitudes were adjacent or at least located close to each other. AL were abandoned 10 to 30 years ago and had since been covered with herbaceous species (mainly Poaceae spp.). C1–4 and PF1 and PF2 were converted 7 to 8 years ago and more than 30 years ago, respectively. PF3, CF1 and CF2, and G1 were located on the natural slope.Twenty-five sampling sites were selected ranging from 381 to 655 m above sea level (asl.) on the slope . These wIn terms of differences in the altitudes of the sampling sites, no appreciable feature was observed in the properties of soils from 0–15 cm. The soil texture class was silty clay loam or finer because the parent materials were derived from the green rock. The cation exchange capacity (CEC) was high . AlthougCyanobacteria and Acidobacteria, respectively. Sequence identity between the two bands was 84%. Soils of the same land use contained very similar bacterial communities regardless of the altitude. This was consistent with the similar soil properties observed at different altitudes. The DGGE profiles of amplified 16S rRNA genes from the soils of natural slopes, including PF3, B1–3, G1, and CF1–2, were similar to each other  in combination with a bead beater at 5,500 rpm for 45 s . The V3 region of the 16S rRNA gene was amplified by the universal primers, 341F-GC and 534R , with NEch other .The pyrosequencing technique was considered to have high levels of robustness, consistency, and resolution . The hypAcidobacteria subdivision 6, which is the most phylogenetically diverse and numerous Acidobacteria subgroup, was the most abundant in all soils tested. The dominancy of an OTU belonging to the class Anaerolineae was greater in PF than in the other land use types. Since the major bacterial community structures were almost similar in each land use regardless of the altitude, we combined the OTU data of each land use from different altitudes. Proteobacteria and Acidobacteria were two major phyla in all the soils examined . The heat map analysis at the genus level showed that the major bacterial community structures were almost similar in P1–6 . This isexamined . Proteobnt in PF . The relreported , 18. Thehat in P . This mabacteria , 22 suppironment  may explIn order to determine differences in the relative diversity and richness of soil bacterial communities among land uses, the observed number of clusters, Chao1 richness, and Shannon diversity index were calculated at a genetic distance of 0.03. Bacterial species richness was slightly higher in PL than in the other land uses . BacteriNo significant differences were noted in bacterial richness or diversity among soil samples; therefore, we attempted to establish whether bacterial community structures differed depending on land use. A beta-diversity analysis was performed using weighted UniFrac distance metrics. In order to assess differences, distances were used for a Principal Coordinate Analysis (PCoA) using QIIME  and visuWe collected soils from terraced P before irrigation and did not analyze the soil bacterial community in flooded P. When bacterial communities in the bulk soil in Japanese P are analyzed from flooded to upland conditions, they are found to remain stable throughout the year . A previThe sequenced read data have been deposited in the DDBJ Sequence Read Archive (DRA) under accession numbers DRA004024 and DRA004025.


Dents were more heat tolerant than Flints. QTL for heat tolerancewith respect to grain yield at field conditions were identified considering multiplepopulations and environments.Hightemperatures have the potential to cause severe damages to maize production. This study aims to elucidate the genetic mechanisms of heat tolerance under field conditions in maize and the genome regions contributing to natural variation. In our study, heat tolerance was assessed on a multi-environment level under non-controlled field conditions for a set of connected intra- and interpool Dent and Flint populations. Our findings indicate that Dent are more heat tolerant during adult stage than Flint genotypes. We identified 11 quantitative trait loci (QTL) including 2 loci for heat tolerance with respect to grain yield. Furthermore, we identified six heat-tolerance and 112 heat-responsive candidate genes colocating with the previously mentioned QTL. To investigate their contributionto the response to heat stress and heat tolerance, differential expression and sequence variation of the identified candidate genes should be subjected to further research.The online version of this article (doi:10.1007/s00122-016-2674-6) contains supplementary material, which is available to authorized users. Zea mays L.) was grown on 184 million hectares in 2013 and was, thus, the second most widely cultivated crop after wheat  propose a measure for heat tolerance which integrates observations from multiple levels of heat stress and assess the heat tolerance of a set of six connected segregating Dent and Flint populations for several traits and on a multi-trait level, (II) identify QTL for heat tolerance with the previously mentioned populations and (III) identify heat-tolerance candidate genes in these QTL regions.1 genotypes  and four Flint  maize inbred lines from the University of Hohenheim  and the KWS Saat AG , comprising two locations with standard conditions in Germany, namely Greven and Einbeck, and two locations with heat conditions, namely Zsombó (Hungary) and Monselice   and female flowering (FF), respectively, were assessed. Furthermore, data for leaf scorching (LS) of young leaves before flowering from 1 (weak damage) to 9 (strong damage) were collected. Total grain fresh yield (FY) was assessed by machine harvesting at physiological maturity, where grain moisture (GM) was measured by near infrared spectroscopy. MF, FF, GM and FY were determined by the respective plant breeding company, LS was assessed by the author of this paper. Grain dry yield per hectare (DY) at 15 % grain moisture was calculated. The anthesis silking interval (ASI) was calculated with  Wilhelm :1\documeThe parental inbred lines of the populations were genotyped with a set of 56,110 single nucleotide polymorphism (SNP) markers using a 50K SNP array  for each trait of the genotypes at each location, the above-mentioned adjusted phenotypic observations were analysed with model  at each AEM across locations with the same condition, i.e. standard and heat, were estimated using model ,4\documeTo calculate AEM of the traits for each location across genotypes and to assess the significance of the condition effect (standard vs. heat conditions), model  was usedHeritability Genotypic j were calculated using model .ng model  with a rp and location j. Therefore, the p and location j  and E the number of replications per locations (2). All mixed model analyses were performed using the software ASReml  was calculated in two steps for each heat-dependent trait  times genotype combination. In the first step, the AEM of the genotypes at each location and the AEM of each location were adjusted by calculating the ratios i and location j withj across all genotypes withi at location j, calculated with model  was adjusted with the HSI for FF (HSIFF) as a cofactor using a linear regression. The residuals of the regression represented the HSI for the adjusted dry yield (HSIDYA).The second step consisted in a stability analysis (cf. Finlay and Wilkinson DY), LS (HSILS), GM (HSIGM), MF (HSIMF) and FF (HSIFF) for all genotypes were used as multi-trait measures for heat susceptibility.For a multi-trait approach, the first two principal components (PC1 and PC2) of a principal component analysis (PCA) considering the previously calculated HSI of the traits DY  ] were estimated on the basis of a 1.5 LOD unit fall.The analyses were performed across all populations  with To test if the dominance effects of each population on the respective QTL were significantly different from 0, significance  from sowing until maturity were between 1400 and 1600 at locations with heat condition and between 900 and 1100 at the two locations with standard conditions  of the respective HSI or PC. With simultaneous fits across all QTL detected for each HSI or PC with several QTL, 19, 17, 19 and 18 % of the variance could be explained for HSIDY, HSIDYA, HSIMF and PC1, respectively. The highest additive effects on QTL for HSIDY and HSIDYA (QHSI:DYa and QHSI:DYb as well as QHSI:DYAa and QHSI:DYAb) were observed for the parental alleles of inbreds P040 and S067, which were the parental inbred lines of population 1. At the genomic position of QHSI:DYa and QHSI:DYAa, the S067 allele had a negative additive effect, whereas at position of QHSI:DYb and QHSI:DYAb, the P040 allele showed a negative additive effect. We observed further a highly significant dominance effect in population 1 for the previously mentioned four QTL, which was negative at QHSI:DYa and QHSI:DYAa and positive at QHSI:DYb and QHSI:DYAb. A total of 6 heat-tolerance genes and 112 heat-responsive genes, identified by Frey et al.  compared to the locations with standard conditions (Einbeck and Greven), except for a heat wave in Germany in late July Table . One weementclass2pt{minimNovel approach to assess heat susceptibility A novel approach to calculate heat susceptibility was applied in our study to combine two characteristics of each genotype, which are involved in its response to heat stress in multiple environments. First, heat susceptibility of each genotype was defined as the difference between observations collected at heat conditions and those collected at standard conditions , we first related the adjusted entry means (AEM) calculated for each genotype at each location to the AEM at the location with least heat stress, i.e. lowest temperature during the entire growing period, in this case the location Einbeck, with 899 of GDD Table . With thHeritability and assessment of traits The heritabilities observed at the location level   Fig. . This co16) Fig.  and, thuMulti-trait measure for heat tolerance The first principal component (PC) of the PC analysis represented a multi-trait measure which combined several HSI to one trait  were observed at both QTL associated with PC1 in population 1, where the parental inbreds were S067 and P040. Heterozygosity for alleles P040 and S067 at position QPC1a and QPC1b resulted in a combined dominance effect of PC1a and QPC1b would improve heat tolerance in terms of grain yield, lower leaf damages produced by heat stress and lead to an increased speed of development enabling plants to escape the strongest summer heat waves.ait Fig. . PC1 expGriffing , who relHeat tolerance of Flint and Dent heterotic pools We observed a lower yield loss and a lower increase of leaf scorching at locations with heat conditions of genotypes derived from Dent × Dent crosses in comparison with genotypes derived from Flint × Flint crosses  were identified, one on chromosome 2 and one on chromosome 3  and between 104 and 141 cM on chromosome 3 (QHSI:DYb and QHSI:DYAb)  were with 0.06 and 0.17 higher than for the other parental alleles   Table , these gCompared to other abiotic stresses associated with climate change (e.g. drought stress), relatively little research has been conducted on heat stress in maize. Existing studies on heat tolerance in maize focussed on a limited number of genotypes with short artificial heat stress events, rather than on the response to heat under field conditions Supplementary material 2 (pdf 23 KB)Below is the link to the electronic supplementary material.


Surgical site infection (SSI) is the second most common infectious complication after urinary tract infection following a delivery by caesarean section (CS). At Bugando Medical Centre there has no study documenting the epidemiology of SSI after CS despite the large number of CSs performed and the relatively common occurrence of SSIs.This was a prospective cohort study involving pregnant women who underwent a CS between October 2011 and February 2012 at Bugando Medical Centre. A total of 345 pregnant women were enrolled. Preoperative, intraoperative and postoperative data were collected using a standardized questionnaire. Wound specimens were collected and processed as per standard operative procedures; and susceptibility testing was carried out using a disc diffusion technique. Data was analyzed using STATA version 11.P = 0.021), severe anaemia , surgical wound class III , multiple vaginal examinations , prolonged duration of operation  and an operation performed by an intern or junior doctor . Staphylococcus aureus was the most common organism (27.3%), followed by Klebsiella pneumoniae (22.7%). Patients with a SSI had a longer average hospital stay than those without a SSI  and the case fatality rate among patients with a SSI was 2.9%.The overall cumulative incidence of SSI was 10.9% with an incidence rate of 37.5 per 10,000 people/day . The median time from CS to the development of SSI was 7 days (interquartile range [IQR] = 6–9 days). Six independent risk factors for post caesarean SSI as identified in this study by multivariate analysis are: hypertensive disorders of pregnancy (HR: 2.5; 95% CI, 1.1-5.6; SSIs are common among women undergoing CSs at Bugando Medical Centre. SSIs were commonly associated with multiple factors. Strategies to control these factors are urgently needed to control SSIs post CS at Bugando Medical Centre and other centres in developing countries. Caesarean section (CS) is the most commonly performed major abdominal operations among women in both developed and developing countries . GloballSSI is a common postoperative complication and is associated with significant morbidity and mortality -7. The oThe occurrence, risk factors and distribution of the pathogens isolated from post CS SSIs at Bugando Medical Centre are not known. Moreover, there is no evidence-based data to support the preoperative and postoperative care protocols at the centre. Therefore this study was carried out to determine the incidence, susceptibility pattern of the isolates and factors associated with SSI among pregnant women undergoing CS at Bugando Medical Centre. The results of this study are necessary to develop an evidence-based treatment protocol for post CS SSIs in this clinical setting and other settings with similar problems.This was a prospective cohort study to determine the incidence, predictors and patterns of surgical site infections among patients undergoing a CS at the Bugando Medical Centre(BMC) between October 2011 and February 2012. At BMC, no routine operation theatre (OT) and sterile services check up; however OT is undergoing major cleanliness once per week and whenever dirty case has been operated. All autoclaves are monitored using chemical and physical indicators daily.All patients who delivered by CS at Bugando Medical Centre and consented for the study were included in the study. Patients who failed to provide information and those who died during the procedure or immediately after CS were excluded from the study.Patients were serially enrolled until the sample size was reached. The sample size was calculated using the Schlesselman formula of the cohort study . The occData was collected using a standardized questionnaire. The data collected included patient, pre- /intra- and postoperative information such as demographic data, wound characteristics and laboratory investigations. Data collected included patient characteristics such as age, parity, co-morbidities such as BMI, DM, and hypertension; pre-operative data such as labour characteristics, preoperative hospital stay and preoperative skin preparation. Intra-operative data collected were type of caesarean section, duration of operation, extent of surgical wound contamination, surgical techniques, blood loss volume, antibiotic prophylaxis and type of anaesthesia used.Surgical wounds were inspected at the time of dressing on day 3 then on day 7 when stitches are removed and thereafter telephone calls were used in every 4 days to enquire for development of SSI and all patients were re-examined at day 30 post operatively Surgical site infection was defined as per Centres for Disease Control and Prevention’s National Nosocomial Infections Surveillance System as described below . SuperfiDeep incisional SSI was defined as infection which occurs within 30 days after the operation and infection involves deep soft tissue  of the incision and at least one of the following: Purulent drainage from the deep incision but not from the organ/space component of the surgical site, OR a deep incision spontaneously dehisces or is deliberately opened by a surgeon when the patient has at least one of the following signs or symptoms: fever (>38C), localized pain, or tenderness, unless site is culture-negative, OR an abscess or other evidence of infection involving the deep incision is found on direct examination, during re-operation, or by histopathologic or radiological examination, OR diagnosis of a deep incisional SSI by a surgeon or attending physician.Organ/space SSI infection was defined as infection occurs within 30 days after the operation and infection involves any part of the anatomy (e.g. organs or spaces), other than the incision, which was opened or manipulated during an operation and at least one of the following: Purulent drainage from a drain that is placed through a stab wound into the organ/space OR organisms isolated from an aseptically obtained culture of fluid or tissue in the organ/space OR an abscess or other evidence of infection involving the organ/space that is found on direct examination, during re-operation, or by histopathologic or radiological examination OR diagnosis of an organ/space SSI by a surgeon or attending physician.Specimens were taken from all patients who clinically were suspected to have surgical site infections. Before sample collection surrounding skin was cleaned with 70% alcohol . Exudates were obtained from the open discharging wounds with a sterile cotton swab or with a syringe . Swabs oAll specimens were processed in accordance with the standard operating procedures of the laboratory. Briefly specimens were inoculated on Blood agar and Mackonkey agar  and incubated aerobically for 24–48 hours. Identification of bacteria was done using conventional physiological and biochemical methods. Biochemical and physiological methods included; gram stain, catalase reaction, coagulase reaction, haemolytic activity on 5% sheep blood agar plate, hippurate hydrolysis and CAMP test for Gram positive bacteria while for gram negative; colonies morphology on blood agar and MacConkey agar, triple sugar iron (TSI) reaction, indole, motility, citrate, urease and hydrogen sulphide production  were used ,18. AntiData collected was entered into a computer using Epidata version 3.1  and analyzed using STATA version 11 . Data was summarized in the form of proportions and frequent tables for categorical variables. Medians with standard deviation or medians with interquartile range were used to summarize continuous variables. A chi-square test was performed to test for significant associations between the predictor and outcome variables within the categorical variables and for continuous variables a Student t-test was performed. Hazard ratio (HR) with 95% confidence interval (CI) was calculated to test for the strength of association between predictor variables and SSIs using univariate analysis followed by multivariate Cox regression analyses for all predictors found to be significant on the univariate analysis. Significant association was defined as a p-value of less than 0.05.Catholic University of Health and Allied Sciences /Bugando Medical Centre review board cleared the study and informed consents were obtained from patients.Twenty six percent of the patients who developed an SSI after discharge did not return for specimen collection. No anaerobic culture was done and lack of antibiotic policy may have affected the SSI rate.A total of 3250 deliveries were performed during the study period, 2476 (76.2%) by the vaginal route and 774 (23.8%) by CS Figure . A totalThe age of the women ranged from 14 to 44 years with a mean of 26.8 ± 5.8 years. The majority of women  were between 20 to 34 years of age and 218 (63.3%) were multiparas. The gestational age at caesarean delivery ranged between 28–44 weeks and a total of 268 neonates (79.8%) were delivered at term. Scars as indication of previous CS were found in 106 (30.7%) patients  and an incidence rate of 37.5 per 10,000 people/day . Types of SSI found were superficial SSI , organ space  and deep SSI . All SSIs occurred among patients who had emergency CSs. The rate of SSI was 12/202(5.9%) for clean wounds and 22/143(29.4%) for clean-contaminated wounds (p = 0.028 Fischer exact test). Most SSIs occurred among patients who had a vertical skin incision .P < 0.0001). The majority of patients with SSIs (n = 23/ 67.6%) were readmitted and one patient with an organ/space SSI died 2 days post relaparotomy due to septicemia.All SSIs occurred between the 3rd and 22nd day postoperatively with a median time of occurrence of 7 days post operation (IQR 6–9 days). Patients with a SSI had longer hospital stays than those without a SSI , higher ASA physical status class , prolonged duration of labor , rupture of membranes prior to surgery lasting 8 hours or longer,, 3 or more vaginal examinations , higher wound class prior to surgery , vertical skin incision , prolonged duration of operation  , and a procedure performed by an intern or junior doctor  ; contaminated wound , multiple vaginal examinations , prolonged duration of operation  and an operation performed by an intern or junior doctor   risk index of 1 to a HR of 9.1 in patients with a NNIS System risk index of 2.Almost all  women received antibiotics as prophylaxis with different timings of administration either before or after skin incision. No specific policy was followed, and the choice of antibiotics used was based on indication of CS and surgeons preference. The antibiotics given could be divided into ampicillin based regimens (170/344) and non-ampicillin based regimen combinations (174/344). A higher SSI rate was observed among patients which received the ampicillin based regimen (11.5%) compared with those which received non-ampicillin based regimen .Pus swabs for aerobic culture and sensitivity were collected for 25 (73.5%) of the clinically suspected postoperative infection cases. Of 25 aerobic cultures, 18 (72.0%) were culture positive and 4 had significant polymicrobial infections, resulting in a total of 22 bacterial isolates.Staphylococcus aureus was the most common organism . Of Staphylococcus aureus, 5 (83.3%) were methicillin sensitive Staphylococcus aureus (MSSA) and 1 (16.7%) was methicillin resistant Staphylococcus aureus (MRSA). Other isolates include Klebsiella spp , Escherichia coli , Acinetobacter spp , Pseudomonas spp , Proteus spp , Enterobacter spp , Micrococcus spp  and coagulase negative Staphylococcus spp .Among the etiological agents isolated, 14 (63.6%) were gram-negative bacilli. Staphylococcus aureus.The majority of gram-positive bacteria isolated were 100% sensitive to vancomycin and imipenem and 85% sensitive to cefazolin. Most of the enteric gram-negative bacteria were highly resistant to ampicillin (100%), amoxicillin/clavulanate (93%), sulphamethaxazole/trimethoprim (78.5%), fosfomycin (70%), tetracycline (70%) and gentamicin (35.7%). They were 100%, 85.7%, 85.7% and 78% sensitive to meropenem, ceftazidime, ciprofloxacin and ceftriaxone respectively. A total of 2 (13%) of the enteric gram-negative bacteria were found to produce extended spectrum beta lactamases (ESBL). Out of 8 organ infections, 5 (62.5%) were due to gram-negative enteric bacteria and 29.4% (5/17) of incisional SSIs were caused by As reported in many other studies in the developing countries ,21,the iEnteric gram-negative bacteria have previously been reported to be associated with severe SSI ,28 . In Various risk factors have been found to predict post caesarean SSI ,9,26. OnIn contrast to previous studies ,29, the ASA physical status classification score of 3 or more was significantly found to predict SSI (p = 0.005), in agreement with previously reported findings ,32. ProlAnother important risk factor for SSI is the absence or delay of antibiotic prophylaxis. In this study there was no standard policy of antibiotic prescription. The choice of antibiotic administered depends on the surgeon and indication of CS. The antibiotics given in this study could be divided into two groups; an ampicillin based regimen and a non-ampicillin based regimen. The timing of antibiotic administration was not consistent. In contrast to previous studies ,34 no siIn this study emergency procedures were a strong predictor of SSI. This finding could be due to sampling bias because approximately 92.5% of CSs were done on an emergence basis. Vertical incision was significantly found to predict SSI; women with vertical skin incisions had a 3.6 fold risk of developing a SSI compared to those with transverse skin incision, and this has been observed before . ProlongEscherichia coli, group B streptococcus (GBS) and Enterococcus spp, or with nasopharyngeal flora such as Staphylococcus aureus or skin flora such Staphylococcus epidermidis[Staphylococcus aureus has been found to be the most common cause of SSI post CS [Escherichia coli, Klebsiella spp., Pseudomonas aeruginosa, Enterobacter spp., Proteus spp. and Enterococcus spp., show a variable distribution pattern [Staphylococcus aureus was the most common isolate taken from superficial skin infections followed by Klebsiella pneumoniae in deep and organ infections. As described in other studies most of Staphylococcus aureus isolated from SSIs were MSSA and only 1 isolate (16.7%) was found to be MRSA. The prevalence of MRSA is similar to the previous study carried out in the same hospital in 2009 [Staphylococcus aureus isolates were resistant to ampicillin, co-trimoxazole and erythromycin; similar to the findings of Fantahamu et al.[The microbial etiology of post CS SSIs has been shown to be diverse, being associated with both vaginal microorganisms such as idermidis. Staphyl post CS ,37. Othe pattern . A similmu et al.. The ratStaphylococcus aureus and Klebsiella pneumoniae which was an ESBL producer. Patients with a SSI had prolonged hospitalization compared with those without a SSI (12.7 vs 4.1 days), similar to the results observed by Killian et al.[As reported previously, the majority of gram-negative enteric bacteria were highly resistant to ampicillin, amoxycillin/clavulanate, tetracycline and co-trimoxazole . A totalan et al..Hypertensive disorders of pregnancy, contaminated wound, multiple vaginal examinations, operations carried out by an intern or junior doctor and prolonged duration of the surgical procedure (longer than 60 minutes) have been found to be independent factors which increase the risk of SSI at Bugando Medical Centre. Identifying high-risk patients who require intensive postoperative care is critical in order to reduce the incidence of SSIs. This can be achieved if independent risk factors for SSI are well understood within the given clinical setting.The authors declare that they have no competing interests.FJM; participated in collecting specimens, collecting clinical data and follow up of the patients and data analysis, SEM; participated in the design and execution of the work, performed microbiological procedures, data analysis, interpretation of data and preparation of the manuscript, MM: Perform data analysis and writing the manuscript; BRK; participated in data analysis, BG; designed the study, collected clinical data and participated in manuscript writing, CI; Designed the study, participated in the interpretation of the data and manuscript writing. All authors have read and approved the final manuscript.