Screening of polymorphisms in the folate pathway in Turkish pediatric Acute Lymphoblastic Leukemia patients

Background and aim: Folate metabolic pathway plays a significant role in leukemogenesis because of its necessity for nucleotide synthesis and DNA methylation. Folate deficiency causes DNA damage. Thus polymorphisms of folate-related genes may affect the susceptibility to childhood Acute Lymphoblastic Leukemia (ALL). MTHFR (Methylenetetrahydrofolate Reductase), DHFR (Dihydrofolate reductase), CBS (Cystathionine b-synthase) and TYMS (Thymidylate Synthase) have an important role in folate pathway because their activated variants modulate synthesis of DNA and levels of folate. In this study, we aimed to investigate whether polymorphisms in genes related to folate metabolic pathway influence the risk to childhood ALL.Subject and methods: The patient groups who were diagnosed with childhood ALL at Losante Pediatric Hematology-Oncology Hospital and healthy control groups were included in the study. MTHFR 677 CT, MTHFR 1298 A-C, CBS 844ins68, DHFR 19-bp and TYMS 1494del6 polymorphisms were screened. Genotyping of these polymorphisms was performed by Restriction Fragment Length Polymorphism (RFLP) analysis and Real Time Polymerase chain Reaction (Real Time-PCR).Results: In total, we have screened 5 polymorphisms in the studied genes. The results were compared between childhood ALL patients and healthy groups. Genotype frequencies of MTHFR 677 C-T, MTHFR 1298 A-C, CBS 844ins68 and DHFR 19-bp del were similar for childhood ALL patients and healthy groups. However, statistical results showed that TYMS 1494del6 may be associated with ALL pathogenesis (p < 0.001).Conclusion: We showed that TYMS polymorphism (rs2853542) may be associated with ALL pathogenesis. In addition, our results demonstrated that MTHFR, DHFR and CBS do not affect development of leukemia. Our study displays also importance as it is the first screening results to identify association with the studied polymorphisms in Turkish patients with childhood ALL and determination of the frequency in Turkish population

Štetni učinci pušenja tijekom trudnoće na DNA i razine reaktivnih oblika kisika (ROS) u krvi majke i novorođenčeta

Some of the genotoxic/carcinogenic substances or metabolites in cigarette smoke are capable of passing through the placenta and harming a newborn’s health. Smoking is also known as a factor in the formation of oxidative damage and the main mechanism involved in the carcinogenic process. Predetermining this genotoxic risk can be successfully achieved by measuring certain parameters of oxidative stress. The comet assay is considered an important biomarker for the evaluation of genotoxic substances and is effective for detecting DNA damage caused by smoking. This study examined third trimester bloods and the cord blood of 28 actively smoking and 22 non-smoking mothers in terms of DNA damage and oxidative stress parameters. Cu/Zn superoxide dismutase (CuZn-SOD), malondialdehyde (MDA), catalase (CAT), plasma nitrite/nitrates (NO2•/NO3•), selenium-dependent glutathione peroxidase (Se-GPx), Cu, and Zn levels were measured as indicators of oxidative damage. There were no significant increases in DNA damage of the actively smoking pregnant group in comparison with the non-smoking pregnant group, either in the third trimester or cord blood. Oxidative stress parameters of smoker and non-smoker groups were statistically different for MDA (p<0.05), CuZn-SOD (p<0.01), Se-GPx (p<0.05) values while the difference was not significant for NO2•/NO3•, CAT, Zn, and Cu values. The same values were also investigated in cord blood, and only NO2•/NO3• (p<0.01), Se-GPx (p<0.01 and CAT (p<0.001) values were found statistically different. Smoking mothers may have been exposed to more oxidative stress than non-smoking mothers.Pojedine genotoksične/kancerogene supstancije ili metaboliti u cigaretnom dimu mogu proći kroz posteljicu i naštetiti zdravlju novorođenčeta. Pušenje je također poznato kao čimbenik pri nastanku oksidacijskog oštećenja DNA i u procesu kancerogeneze. Ovaj genotoksični rizik može se uspješno odrediti mjerenjem određenih parametara oksidacijskog stresa. Komet-test smatra se važnim biološkim biljegom pri evaluaciji genotoksičnih supstancija i iznimno učinkovitim sredstvom pronalaženja oštećenja DNA uzrokovanih pušenjem. Ova studija proučava krv trudnica u trećem tromjesečju trudnoće i fetalnu krv 28 majki aktivnih pušačica te 22-ju majki nepušačica vezano za oksidacijska oštećenja DNA i parametre oksidacijskog stresa. Razine Cu/Zn superoksidne dismutaze (CuZn-SOD), malondialdehida (MDA), katalaze (CAT), nitrita/nitrata u plazmi (NO2-/NO3-), selenijeve glutation peroksidaze (Se-GPx), Cu i Zn mjerene su kao pokazatelji oksidacijskog oštećenja. Nije bilo značajnih povećanja oštećenja DNA u skupini trudnica aktivnih pušačica u usporedbi sa skupinom trudnica nepušačica, ni u krvi iz trećeg tromjesečja ni u fetalnoj krvi. Parametri oksidacijskog stresa pušačke i nepušačke skupine bili su statistički različiti za vrijednosti MDA (p<0,05), CuZn-SOD (p<0,01), Se-GPx (P<0,05), dok razlika nije bila značajna za vrijednosti NO2-/NO3-, CAT, Zn i Cu. Iste su vrijednosti ispitane i u fetalnoj krvi, a jedino su vrijednosti NO2-/NO3- (p<0,01), Se-GPx (p<0,01) i CAT (p<0,001) bile statistički različite. Vjerojatno je da su majke pušačice bile izložene većem oksidacijskom stresu od majki nepušačica

Türkiye’de bulunan yoğun bakımlarda sabun, kağıt havlu ve alkol bazlı el dezenfektanı yeterli mi?: Phokai çalışması sonuçları

Introduction: Hand hygiene is one of the most effective infection control measures to prevent the spread of healthcare-associated infections (HCAI). Water, soap, paper towel and hand disinfectant must be available and adequate in terms of effective hand hygiene. The adequacy of hand hygiene products or keeping water-soap and paper towel is still a problem for many developing countries like Turkey. In this multicenter study, we analyzed the adequacy in number and availability of hand hygiene products.Materials and Methods: This study was performed in all intensive care units (ICUs) of 41 hospitals (27 tertiary-care educational, 10 state and four private hospitals) from 22 cities located in seven geographical regions of Turkey. We analyzed water, soap, paper towel and alcohol-based hand disinfectant adequacy on four different days, two of which were in summer during the vacation time (August, 27th and 31st 2016) and two in autumn (October, 12th and 15th 2016).Results: The total number of ICUs and intensive care beds in 41 participating centers were 214 and 2357, respectively. Overall, there was no soap in 3-11% of sinks and no paper towel in 10-18% of sinks while there was no alcohol-based hand disinfectant in 1-4.7% of hand disinfectant units on the observation days. When we compared the number of sinks with soap and/or paper towel on weekdays vs. weekends, there was no significant difference in summer. However, on autumn weekdays, the number of sinks with soap and paper towel was significantly lower on weekend days (p<0.0001, p<0.0001) while the number of hand disinfectant units with alcohol-based disinfectant was significantly higher (p<0.0001).Conclusion: There should be adequate and accessible hand hygiene materials for effective hand hygiene. In this study, we found that soap and paper towels were inadequate on the observation days in 3-11% and 10-18% of units, respectively. Attention should be paid on soap and paper towel supply at weekends as well

Self-reported antibiotic stewardship and infection control measures from 57 intensive care units: An international ID-IRI survey

We explored the self-reported antibiotic stewardship (AS), and infection prevention and control (IPC) activities in intensive care units (ICUs) of different income settings. A cross-sectional study was conducted using an online questionnaire to collect data about IPC and AS measures in participating ICUs. The study participants were Infectious Diseases-International Research Initiative (IDI-IR) members, committed as per their institutional agreement form. We analyzed responses from 57 ICUs in 24 countries (Lower-middle income (LMI), n = 13; Upper-middle income (UMI), n = 33; High-income (HI), n = 11). This represented (similar to 5%) of centers represented in the ID-IRI. Surveillance programs were implemented in (76.9%-90.9%) of ICUs with fewer contact precaution measures in LMI ones (p = 0.02); (LMI:69.2%, UMI:97%, HI:100%). Participation in regional antimicrobial resistance programs was more significantly applied in HI (p = 0.02) (LMI:38.4%,UMI:81.8%,HI:72.2%). AS programs are implemented in 77.2% of institutions with AS champions in 66.7%. Infectious diseases physicians and microbiologists are members of many AS teams (59%&amp;50%) respectively. Unqualified healthcare professionals(42.1%), and deficient incentives(28.1%) are the main barriers to implementing AS. We underscore the existing differences in IPC and AS programs' implementation, team composition, and faced barriers. Continuous collaboration and sharing best practices on APM is needed. The role of regional and international organizations should be encouraged. Global support for capacity building of healthcare practitioners is warranted. (C) 2022 Published by Elsevier Ltd on behalf of King Saud Bin Abdulaziz University for Health Sciences

Identifying a Window of Vulnerability during Fetal Development in a Maternal Iron Restriction Model

It is well acknowledged from observations in humans that iron deficiency during pregnancy can be associated with a number of developmental problems in the newborn and developing child. Due to the obvious limitations of human studies, the stage during gestation at which maternal iron deficiency causes an apparent impairment in the offspring remains elusive. In order to begin to understand the time window(s) during pregnancy that is/are especially susceptible to suboptimal iron levels, which may result in negative effects on the development of the fetus, we developed a rat model in which we were able to manipulate and monitor the dietary iron intake during specific stages of pregnancy and analyzed the developing fetuses. We established four different dietary-feeding protocols that were designed to render the fetuses iron deficient at different gestational stages. Based on a functional analysis that employed Auditory Brainstem Response measurements, we found that maternal iron restriction initiated prior to conception and during the first trimester were associated with profound changes in the developing fetus compared to iron restriction initiated later in pregnancy. We also showed that the presence of iron deficiency anemia, low body weight, and changes in core body temperature were not defining factors in the establishment of neural impairment in the rodent offspring

Development of Antimicrobial Therapies Against Campylobacter jejuni in Poultry

Campylobacter jejuni is a Gram negative bacterium and well-recognised as a major etiologic agent for acute gastro-intestinal diseases in humans, particularly in the elderly and in children. C. jejuni can be isolated from a variety of wild and domestic animals, especially poultry and is primarily transmitted through uncooked or raw foods of animal origin. The consumption and mishandling of poultry products is the main cause of this illness in humans, but no effective control measures are available at present. Development of strategies to control colonisation of Campylobacter in poultry may be effective in reducing the incidence of this disease. The major aim of the present study is to improve food safety in the chicken industry by the reduction or prevention of C. jejuni colonization and carriage in poultry. To achieve this, the present study has focused on the antimicrobial potential of Australian native plants and plant derived compounds for their potent feed additives for their potential in reducing colonization and carriage of C jejuni in poultry. In addition, selected phage-displayed peptides were also tested for potential antimicrobial activity against C. jejuni. Antimicrobial activity of 115 extracts from 109 Australian plant species was screened against two C. jejuni strains using an in vitro broth microdiluton assay. The effects of plant extracts against various phases of C. jejuni growth were compared. C. jejuni cells were more susceptible in the exponential growth phase compared to the stationary phase. Most of the plant extracts (93%) showed activity at a concentration between 32 and 1024 μg/mL against at least one C. jejuni strain. Seventeen plant extracts that had activity at concentration less than 256 μg/mL were selected for further testing against six additional C. jejuni strains, as well as Campylobacter coli, and also against a panel of enteric gut pathogens (Escherichia coli, Salmonella typhimurium, Bacillus cereus, Proteus mirabilis and Enterecoccus faecalis). The best antimicrobial activity was obtained from the extract of Eucalyptus occidentalis, with an inhibitory concentration of 32 μg/mL against C. jejuni and B. cereus. In addition, three essential oils (EOs) and five terpenoid compounds were examined for their antimicrobial potential against two representatives of C. jejuni strains, C. coli and the other gut bacteria. Antimicrobial activity was determined by the use of disc diffusion and broth dilution techniques. Additionally, the antimicrobial activity of neem oil (Azadirachta indica) in different formulations with some of these active components was investigated for its synergistic activity towards the same bacteria in disc diffusion assay. The essential oil from Melaleuca alternifolia showed the most prominent activity against all bacteria with inhibitory concentrations in the range 0.001-0.25% v/v. In these in vitro studies, Campylobacter spp. was found to be the most susceptible organism. The usefulness of in vitro fermentation technique to test antimicrobial activity of natural compounds towards C. jejuni in mixed culture was also explored. Mixed caecal bacteria spiked with C. jejuni were incubated in anaerobic media for 48 h with chicken feed as the substrate. The number of C. jejuni was determined and gas production was recorded throughout the incubation period. At the end of the experiment, methane and volatile fatty acids (VFAs) concentration was determined as well. Test agents, which reduced the number of C. jejuni had no adverse effects on total gas and VFAs production, were selected for in vivo trial. Phage display technology was also used to identify antimicrobial peptides that would inhibit C. jejuni. This technique allows the peptides and proteins to be displayed on the surface of filamentous phage by inserting them into phage major and minor coat proteins. Using this approach, seven ligands with 15-mer peptide interacting with C. jejuni cells were selected and tested to determine their antimicrobial activity against C. jejuni. All phage peptides had high growth-inhibitory potency against C. jejuni. Among them, DT 3/15 showed particularly a high bactericidal activity in in vitro assay. The peptides were also tested towards the other gut bacteria given above and did not show any activity. In addition, the effect of DT/15 on the fermentation kinetics, C. jejuni counts and VFAs production was monitored using an in vitro fermentation method. DT3/15 had no effects on fermentation parameters and no inhibition of C. jejuni. Regardless of the results obtained in vitro fermentation assay, broth dilution study demonstrated that the isolated clones displayed high specificity against Campylobacter cells as no activity was observed on a panel of other bacteria. The use of plant derived agents as C. jejuni inhibitor in chicken feed was tested in an animal trial. Over a seven-week, birds were fed a normal basal diet or basal diet supplemented with the plant extracts derived from Acacia decurrens and Eremophila glabra, lemon myrtle oil, terpinene-4-ol, formulation of compounds named α-tops and antibiotic, virginiamycin. The number of C. jejuni was determined by using the traditional culture and RTQ-PCR methods from the faecal and caecal samples. In addition, body weight gain (BWG) and feed intake (FI) were recorded weekly, and feed conversion efficacy (FCE) were calculated. The mean log10 counts of C. jejuni tended to be lower in faecal and caecal samples obtained from α-tops supplemented group than other treatments and control diet. However, other supplementations did not (P>0.05) cause differences in C. jejuni numbers, even though observably less C. jejuni was counted in the first faecal shedding. No difference (P>0.05) in broiler performance (BWG, FI and FCE) were obtained for dietary supplementation, except E. glabra extracts which had negative impact (

Insufficient decontamination in Sewage treatment plants induce the risk of artificial selection of extended-spectrum beta-lactamase producing Escherichia coli

Quantitative data about extended-spectrum beta-lactamase producing E. coli ( ESBLEC) in the wastewaters are scarce, especially in developing countries. These data could be useful to raise awareness about the potential risk of spreading ESBLEC strains in the community. Water samples were collected weekly over a 10-week period, from one urban sewage treatment plant(STP), one rural STP and one hospital complex's wastewater(HWW) in Turkey. Mean E. coli and ESBLEC loads were determined for each sampling point. For the 580 ESBLEC isolated, antimicrobial resistance profiles, phylogenetic grouping, presence of common beta-lactamese-typesand integrons were studied using PCR. The mean ESBLEC ratio was accounted for 0.58%, 0,12%, 1.53% of the total E. coli in urban, rural untreated wastewater and HWW, respectively. These values were higher for the outlets. The mean number of different antimicrobial classes to which the strains were resistant was highest in urban STP(4.0 +/- 1.6). The antimicrobial resistance ratios of ESBLEC strains isolated from HWW were observed to be in between those of urban and rural STPs. The most common phylogenetic group was C composing(29.7%) and the most susceptible strains belonged to phylogroup B1. Wastewater treatments without sufficient decontamination, resulting in artificial selection of ESBLEC might lead to public health risk as these strains reach communities through environment. To avoid such risks and protect the human health as well as the environment, well-established decontamination measures imposing barriers against this artificial selection should be implemented