Analysis of the Genome and Mobilome of a Dissimilatory Arsenate Reducing Aeromonas sp. O23A Reveals Multiple Mechanisms for Heavy Metal Resistance and Metabolism

Aeromonas spp. are among the most ubiquitous microorganisms, as they have been isolated from different environmental niches including waters, soil, as well as wounds and digestive tracts of poikilothermic animals and humans. Although much attention has been paid to the pathogenicity of Aeromonads, the role of these bacteria in environmentally important processes, such as transformation of heavy metals, remains to be discovered. Therefore, the aim of this study was a detailed genomic characterization of Aeromonas sp. O23A, the first representative of this genus capable of dissimilatory arsenate reduction. The strain was isolated from microbial mats from the Zloty Stok mine (SW Poland), an environment strongly contaminated with arsenic. Previous physiological studies indicated that O23A may be involved in both mobilization and immobilization of this metalloid in the environment. To discover the molecular basis of the mechanisms behind the observed abilities, the genome of O23A (∼5.0 Mbp) was sequenced and annotated, and genes for arsenic respiration, heavy metal resistance (hmr) and other phenotypic traits, including siderophore production, were identified. The functionality of the indicated gene modules was assessed in a series of minimal inhibitory concentration analyses for various metals and metalloids, as well as mineral dissolution experiments. Interestingly, comparative analyses revealed that O23A is related to a fish pathogen Aeromonas salmonicida subsp. salmonicida A449 which, however, does not carry genes for arsenic respiration. This indicates that the dissimilatory arsenate reduction ability may have been lost during genome reduction in pathogenic strains, or acquired through horizontal gene transfer. Therefore, particular emphasis was placed upon the mobilome of O23A, consisting of four plasmids, a phage, and numerous transposable elements, which may play a role in the dissemination of hmr and arsenic metabolism genes in the environment. The obtained results indicate that Aeromonas sp. O23A is well-adapted to the extreme environmental conditions occurring in the Zloty Stok mine. The analysis of genome encoded traits allowed for a better understanding of the mechanisms of adaptation of the strain, also with respect to its presumable role in colonization and remediation of arsenic-contaminated waters, which may never have been discovered based on physiological analyses alone

Disturbances in angiogenesis and vascular maturation in the skin are associated with diabetic kidney disease in type 1 diabetes

Introduction. The skin, as one of the most accessible tissues, is frequently used for investigations of microcirculation and angiogenesis. The aim of this study was to assess the relationship between the dermal microvessel density (MVD) and maturity and the presence of diabetic kidney disease (DKD) in adults with type 1 diabetes (T1D). Skin as the most accessible organ served as a model for the study of angiogenesis. Materials and methods. 148 consecutive T1D patients (87 men), median age of 41 [interquartile range (IQR): 31–49] years and diabetes duration of 21 (17–30) years, participated in the study. The patients were under the care of the Department of Internal Medicine and Diabetology, Poznan University of Medical Sciences. Diabetic kidney disease was diagnosed in patients with increased albuminuria and at least 10-year duration of diabetes or evidence of diabetic retinopathy. The skin biopsy was performed on distal part of lower leg, using a sterile, disposable 3 mm biopsy punch with plunger (Disposable Biopsy Punches, Integra™ Miltex®). In the immunohistochemical analyses, we used: anti-CD133, anti-CD34, anti-CD31, and anti-von Willebrand factor (vWF) autoantibodies. Microvessel density measurement in all specimens was performed using “hot spots technique”. Slides were scanned using the MiraxMidi scanner (Carl Zeiss) and were viewed using CaseViewer (3DHISTECH Ltd. Budapest, Hungary). Data were analyzed using Statistica v. 13 software. Results. In the study group 21 patients with diagnosis DKD+, as compared to 127 subjects withaout DKD–, had longer duration of diabetes [30 (IQR: 21–36) vs. 21 (16–28) years, p = 0.002], higher prevalence of hypertension [14 (67%) vs. 37 (29%), p = 0.002], lower estimated glomerular filtration rate (eGFR) [66 (55–88) vs. 94 (83–106) mL/min/1.73 m2, p &lt; 0.001]. Median MVD compared between groups with and without DKD, was similar for CD34+ vessels/1 mm2 [123 (100–170) vs. 121 (100–170), p = 0.775], CD133+ vessels/1 mm2 [79 (50–100) vs. 79 (63–93), p = 0.823], and for CD31+ vessels/ 1 mm2 [29 (21–46) vs. 38 (17–58), p = 0.454]. Median MVD vWF+ vessels/1 mm2 was lower in the group with than without DKD: 42 (25–54) vs. 54 (43–71), p = 0.009. The values given above were calculated for both layers of the dermis (papillary and reticular dermis). In multivariate logistic regression analysis presence of diabetic kidney disease was associated with lower median vWF+ MVD [odds ratio: 0.97 (95% confidence interval: 0.95–0.99), p = 0.017], with adjustment for age, gender, eGFR value, diabetes duration and presence of hypertension. MVD did not differ significantly between chronic kidney disease stages. Conclusion. In patients with type 1 diabetes and diabetic kidney disease the disturbances in the angiogenesis and vascular maturation are present. The number of mature blood vessels (vWF+) in the skin is reduced. Disturbances in the angiogenesis occur at early stages of diabetic kidney disease.Introduction. The skin, as one of the most accessible tissues, is frequently used for investigations of microcirculation and angiogenesis. The aim of this study was to assess the relationship between the dermal microvessel density (MVD) and maturity and the presence of diabetic kidney disease (DKD) in adults with type 1 diabetes (T1D). Skin as the most accessible organ served as a model for the study of angiogenesis. Materials and methods. 148 consecutive T1D patients (87 men), median age of 41 [interquartile range (IQR): 31–49] years and diabetes duration of 21 (17–30) years, participated in the study. The patients were under the care of the Department of Internal Medicine and Diabetology, Poznan University of Medical Sciences. Diabetic kidney disease was diagnosed in patients with increased albuminuria and at least 10-year duration of diabetes or evidence of diabetic retinopathy. The skin biopsy was performed on distal part of lower leg, using a sterile, disposable 3 mm biopsy punch with plunger (Disposable Biopsy Punches, Integra™ Miltex®). In the immunohistochemical analyses, we used: anti-CD133, anti-CD34, anti-CD31, and anti-von Willebrand factor (vWF) autoantibodies. Microvessel density measurement in all specimens was performed using “hot spots technique”. Slides were scanned using the MiraxMidi scanner (Carl Zeiss) and were viewed using CaseViewer (3DHISTECH Ltd. Budapest, Hungary). Data were analyzed using Statistica v. 13 software. Results. In the study group 21 patients with diagnosis DKD+, as compared to 127 subjects withaout DKD–, had longer duration of diabetes [30 (IQR: 21–36) vs. 21 (16–28) years, p = 0.002], higher prevalence of hypertension [14 (67%) vs. 37 (29%), p = 0.002], lower estimated glomerular filtration rate (eGFR) [66 (55–88) vs. 94 (83–106) mL/min/1.73 m2, p < 0.001]. Median MVD compared between groups with and without DKD, was similar for CD34+ vessels/1 mm2 [123 (100–170) vs. 121 (100–170), p = 0.775], CD133+ vessels/1 mm2 [79 (50–100) vs. 79 (63–93), p = 0.823], and for CD31+ vessels/ 1 mm2 [29 (21–46) vs. 38 (17–58), p = 0.454]. Median MVD vWF+ vessels/1 mm2 was lower in the group with than without DKD: 42 (25–54) vs. 54 (43–71), p = 0.009. The values given above were calculated for both layers of the dermis (papillary and reticular dermis). In multivariate logistic regression analysis presence of diabetic kidney disease was associated with lower median vWF+ MVD [odds ratio: 0.97 (95% confidence interval: 0.95–0.99), p = 0.017], with adjustment for age, gender, eGFR value, diabetes duration and presence of hypertension. MVD did not differ significantly between chronic kidney disease stages. Conclusion. In patients with type 1 diabetes and diabetic kidney disease the disturbances in the angiogenesis and vascular maturation are present. The number of mature blood vessels (vWF+) in the skin is reduced. Disturbances in the angiogenesis occur at early stages of diabetic kidney disease

Genetic variants in transforming growth factor-β gene (TGFB1) affect susceptibility to schizophrenia

Immense body of evidence indicates that dysfunction of immune system is implicated in the etiology of schizophrenia. The immune theory of schizophrenia is supported by alterations in cytokine profile in the brain and peripheral blood. Given the strong genetic background of schizophrenia, it might be assumed that aberrant production of cytokines might be the consequence of genetic factors. This study aimed at investigating the association between schizophrenia susceptibility and selected functional polymorphisms in genes encoding cytokines including: interleukin-2 (IL2 −330T>G, rs2069756), interleukin-6 (IL-6 −174G>C, rs1800795), interferon-γ (IFNG +874T>A, rs2430561) as well as for the first time transforming growth factor-β1 (TGFB1 +869T>C, rs1800470 and +916G>C, rs1800471). We recruited 151 subjects with schizophrenia and 279 controls. There was a significant difference in the genotype distribution and allelic frequency of the TGFB1 +869T>C between patients with schizophrenia and healthy controls (p < 0.05). The risk of schizophrenia was more than two-fold higher in carriers of T allele (CT+TT genotypes) than individuals with CC genotype. Given documented gender differences in incidence of schizophrenia, we conducted separate analyses of male and female participants. We have shown that the association was significant in females, while in males it reached a trend toward statistical significance. To the best of our knowledge, it is the first report showing the association between TGFB1 +869T>C polymorphism and schizophrenia

Is there respectful maternity care in Poland? Women’s views about care during labor and birth.

Background: Abuse against women in labor starts with subtle forms of discrimination that can turn into overt violence. Therefore it is crucial to work towards prevention and elimination of disrespect and ill-treatment in medical facility perinatal care in which staff allows such abuse. The aim of the study was to analyze the experiences of women related to perinatal care. Special emphasis was put on experiences that had traits indicating disrespectful and offensive care during childbirth in medical facilities providing perinatal care. Methods: This was a cross-sectional survey. A questionnaire was prepared for respondents who gave birth in medical facilities. Information about the study was posted on the website of a non-governmental foundation dealing with projects aimed at improving perinatal care. The respondents gave online consent for processing the submitted data. 8378 questionnaires were submitted. The study was carried out between February 06 and March 20, 2018. The results were analyzed using the Chi-square independence test. The analysis was carried out at the significance level of 0.05 in Excel, R and SPSS. Results: During their hospital stay, 81% of women in the study experienced violence or abuse from medical staff on at least one occasion. The most common abuse was having medical procedures without prior consent. Inappropriate comments made by staff related to their own or a woman’s situation were reported in 25% of situations, whilst 20% of women experienced nonchalant treatment. In the study 19.3% of women reported that the staff did not properly care for their intimacy and 1.7% of the respondents said that the worst treatment was related to feeling anonymous in the hospital. Conclusions: The study shows that during Polish perinatal care women experience disrespectful and abusive care. Most abuse and disrespect involved violation of the right to privacy, the right to information, the right to equal treatment, and the right to freedom from violence. The low awareness of abuses and complaints reported in the study may result from women’s ignorance about relevant laws related to human rights

Uridylation by TUT4/7 Restricts Retrotransposition of Human LINE-1s

LINE-1 retrotransposition is tightly restricted by layers of regulatory control, with epigenetic pathways being the best characterized. Looking at post-transcriptional regulation, we now show that LINE-1 mRNA 3′ ends are pervasively uridylated in various human cellular models and in mouse testes. TUT4 and TUT7 uridyltransferases catalyze the modification and function in cooperation with the helicase/RNPase MOV10 to counteract the RNA chaperone activity of the L1-ORF1p retrotransposon protein. Uridylation potently restricts LINE-1 retrotransposition by a multilayer mechanism depending on differential subcellular localization of the uridyltransferases. We propose that uridine residues added by TUT7 in the cytoplasm inhibit initiation of reverse transcription of LINE-1 mRNAs once they are reimported to the nucleus, whereas uridylation by TUT4, which is enriched in cytoplasmic foci, destabilizes mRNAs. These results provide a model for the post-transcriptional restriction of LINE-1, revealing a key physiological role for TUT4/7-mediated uridylation in maintaining genome stability

Malnutrition, inflammation, atherosclerosis syndrome (MIA) and diet recommendations among end-stage renal disease patients treated with maintenance hemodialysis

Malnutrition-inflammation-atherosclerosis syndrome is one of the causes of increased mortality in chronic kidney disease (CKD). The aim of the study was to assess the inflammation and nutritional status of patients in end-stage kidney disease treated with maintenance hemodialysis. The study included a group of 98 hemodialyzed patients with stage 5 CKD (38 women and 60 men). Albumin, prealbumin (PRE), and C-reactive protein (CRP) were measured in serum samples collected before mid-week dialysis. Fruit and vegetables frequency intakes were assessed with a questionnaire. CRP was above the reference limit of 5 mg/L in 53% of patients. Moreover, the Glasgow Prognostic Score (GPS) indicated the co-occurrence of inflammation and protein calorie malnutrition in 11% of patients, and the presence of either inflammation or malnutrition in 25%. The questionnaire revealed that hemodialyzed patients frequently exclude fruit and vegetables from their diets. Nearly 43% of the interviewed patients declared frequently eating vegetables, and 35% declared frequently eating fruit, a few times per week or less. The most frequently selected fruit and vegetables had a low antioxidant capacity. The strict dietary restrictions in CKD are difficult to fulfill, and if strictly followed, may lead to protein-calorie malnutrition

Effects of interactions between variation in dopaminergic genes, traumatic life events, and anomalous self-experiences on psychosis proneness : results from a cross-sectional study in a nonclinical sample

Background: there is a growing number of studies showing interactions between genetic polymorphisms associated with dopaminergic neurotransmission and traumatic life events (TLEs) on a risk of psychotic-like experiences (PLEs). Anomalous self-experiences (ASEs) have been associated both with TLEs as well as with PLEs. However, it remains unknown what is the role of ASEs in the complexity of gene - environment interactions on the emergence of PLEs. Patients and methods: we included 445 young adults - university students from three big cities in Poland. We used the Traumatic Events Checklist to assess TLEs, the Inventory of Psychotic-Like anomalous self-experiences in order to measure ASEs, and the Prodromal Questionnaire (PQ16) to record the level of PLEs. The following gene polymorphisms, related to dopaminergic neurotransmission, were determined: the catechol-O-methyltransferase (COMT) rs4680 polymorphism, the dopamine D2 receptor (DRD2) rs6277 polymorphism, and the dopamine transporter 1 (DAT1) rs28363170 polymorphism. Results: there was a significant effect of the interaction between the DAT1 polymorphism, a severity of ASEs, and a history of TLEs on the level of PLEs. Among the DAT1 10R/10R homozygotes with low level of ASEs, a severity of PLEs was significantly higher in individuals with a history of any TLEs. Higher scores of the PQ16 were associated with a greater severity of ASEs both in the DAT1 9R allele carriers and the DAT1 10R/10R homozygotes. Conclusion: our findings imply that genetic liability related to aberrant dopamine transport might impact the association between TLEs and PLEs in subjects with high levels of ASEs

Częstość występowania zespołu rozrostu bakteryjnego jelita cienkiego (SIBO) u pacjentów z cukrzycą

Introduction. Gastrointestinal symptoms are present in 50–70% of patients with diabetes. Small intestinal bacterial overgrowth (SIBO) is expansion of microorganisms colonizing the large intestine into the small intestine, leading to disorders of digestion and ab­sorption. Material and methods. The study included 200 patients (91 with type 1 and 109 with type 2 diabetes hospital­ized in years 2013–2015 in the Department of Internal Medicine and Diabetology, Poznan University of Medi­cal Sciences. The control group consisted of 20 healthy volunteers. Hydrogen breath tests with Gastro+ Gastrolyzer (Bedfont) were used to assess SIBO. After drinking 20 g of lactulose we checked exhaled hydro­gen concentration. The result was considered positive when exhaled hydrogen of first breath was ≥ 20 ppm (parts per milion) or an increase in the output value of the exhaled hydrogen was 12 ppm during the first 60 minutes of the test. Results. We observed lower prevalence of SIBO in the test group as compared to controls [82 patients (41%) vs. 15 patients (75%); p = 0.0043]. Moreover, patients with type 1 diabetes as well as type 2 diabetes had lower incidence of SIBO as compared to controls [accordingly, 42% vs. 75%, p=0.008, 40% vs. 75%, p=0.006]. In the logistic regression model this relationship was independent of age, gender and BMI (OR: 5,57, 95% CI: 1,78–17,47; p = 0,003). Conclusion. The prevalence of microbial proliferation of the small intestine in patients with diabetes is lower than in healthy subjects. One possible cause might be the beneficial effect of nutritional therapy in patients with diabetes.Wstęp. Dolegliwości ze strony przewodu pokarmo­wego mogą występować u 50–70% osób z cukrzycą. Zespół rozrostu bakteryjnego (SIBO) to nadmierny rozplem w obrębie jelita cienkiego drobnoustrojów kolonizujących jelito grube, prowadzący do zaburzeń trawienia i wchłaniania. Celem pracy była ocena częstości występowania SIBO u pacjentów z cukrzycą, a także związek występowania SIBO z wyrównaniem metabolicznym cukrzycy i obecnością przewlekłych powikłań choroby. Materiał i metody. Grupę badaną stanowiło 200 osób z cukrzycą (typu 1 — 91 osób; typu 2 — 109 osób), ho­spitalizowanych w latach 2013–2015 w Klinice Chorób Wewnętrznych i Diabetologii Uniwersytetu Medycz­nego im. Karola Marcinkowskiego w Poznaniu. Grupę kontrolną stanowiło 20 zdrowych ochotników. Obec­ność rozrostu bakteryjnego oceniano w nieinwazyjnym teście oddechowym przy użyciu urządzenia Gastro+ Gastrolyzer (firmy Bedfont) z wykorzystaniem 20 g laktulozy rozpuszczonej w 200 ml wody, z oceną stężenia wodoru w wydychanym powietrzu. Za wynik dodatni uznano zawartość wydychanego wodoru ≥ 20 ppm (parts per milion — części na milion) lub wzrost wartości wyjściowej wydychanego wodoru o 12 ppm w czasie pierwszych 60 minut badania. Wyniki. Występowanie SIBO w grupie badanej stwierdzono u 82 pacjentów (41%), natomiast w grupie kontrolnej u 15 osób (75%), p = 0,0043. Istotną różnicę stwierdzono również w oddzielnym porównaniu osób z cukrzycą typu 1 i 2 i z grupą kontrolną. Obecność SIBO stwierdzano istotnie częściej u osób zdrowych (75%) niż u osób z cukrzycą typu 1 (42%, p = 0,008) oraz typu 2 (40%, p = 0,006). W modelu regresji logistycznej obecność cukrzycy wiązała się z rzadszym występowaniem SIBO niezależnie od wieku, płci i wartości BMI (OR: 0,18, 95% CI: 0,06–0,56; p = 0,003). Wnioski. U pacjentów z cukrzycą częstość występo­wania zespołu rozrostu bakteryjnego jelita cienkiego jest mniejsza niż u osób zdrowych. Jedną z możliwych przyczyn stwierdzonego związku może być korzystny wpływ leczenia żywieniowego u osób z cukrzycą