The Transcriptomic and Bioinformatic Characterizations of Iron Acquisition and Heme Utilization in Avibacterium paragallinarum in Response to Iron-Starvation

Avibacterium paragallinarum is the pathogen of infectious coryza, which is a highly contagious respiratory disease of chickens that brings a potentially serious threat to poultry husbandry. Iron is an important nutrient for bacteria and can be obtained from surroundings such as siderophores and hemophores. To date, the mechanisms of iron acquisition and heme utilization as well as detailed regulation in A. paragallinarum have been poorly understood. In this study, we investigated the transcriptomic profiles in detail and the changes of transcriptomes induced by iron restriction in A. paragallinarum using RNA-seq. Compared with the iron-sufficiency control group, many more differentially expressed genes (DEGs) and cellular functions as well as signaling pathways were verified in the iron-restriction group. Among these DEGs, the majority of genes showed decreased expression and some were found to be uniquely present in the iron-restriction group. With an in-depth study of bioinformatic analyses, we demonstrated the crucial roles of the Hut protein and DUF domain-containing proteins, which were preferentially activated in bacteria following iron restriction and contributed to the iron acquisition and heme utilization. Consequently, RT-qPCR results further verified the iron-related DEGs and were consistent with the RNA-seq data. In addition, several novel sRNAs were present in A. paragallinarum and had potential regulatory roles in iron homeostasis, especially in the regulation of Fic protein to ensure stable expression. This is the first report of the molecular mechanism of iron acquisition and heme utilization in A. paragallinarum from the perspective of transcriptomic profiles. The study will contribute to a better understanding of the transcriptomic response of A. paragallinarum to iron starvation and also provide novel insight into the development of new antigens for potential vaccines against infectious coryza by focusing on these iron-related genes.

Do We Need Fluid Restriction After Stapled Hemorrhoidopexy? A Pilot, Double-Blinded, Randomized Controlled Trial

Background: Conventional hemorrhoidectomy leads to a high urinary retention rate and fluid restriction is commonly recommended to minimize complications. However, the need for postoperative fluid restriction among patients who have undergone stapled hemorrhoidopexy is unclear. We aimed to determine whether fluid restriction after stapled hemorrhoidopexy with/without partial external hemorrhoidectomy could reduce urinary retention and postoperative pain.Patients And Methods: In this prospective, double-blinded, randomized controlled trial, we enrolled patients who had grade III or IV hemorrhoids and underwent stapled hemorrhoidopexy with/without partial external hemorrhoidectomy; 250 mL/12 h of sterile 0.9% saline was administered to the fluid restriction group after the operation, and 1000 mL/12 h was given to the non-fluid-restriction group. We focused on the need for fluid restriction after stapled hemorrhoidopexy.Results: Fifty patients were studied in two groups. The surgical outcomes of the two groups were no different. The median subjective pain scores at 0, 8, 16, 24 h after the operation showed no significant difference between the two groups (p = 0.55 at 0 h; p = 0.38 at 8 h; p = 0.98 at 16 h; and p = 0.66 at 24 h). The mean times to first urination after the operation were 700.04 ± 455.03 min in the non-fluid-restriction group and 737.16 ± 426.32 min in the fluid-restriction group (no significant difference; p = 0.67). No postoperative urinary retention was found in either group of patients.Conclusions: Fluid restriction after stapled hemorrhoidopexy is unnecessary.Trial registration: This study was reviewed and approved by the Tri-Service General Hospital Institutional Review Board for human subjects (No. 2-106-05-063). This study also had trial registration with clinicalTrials.gov (Identifier: NCT04459039).

Pengaruh Kombinasi Suplementasi Omega 3 dengan Kalori Restriksi Terhadap Penyembuhan Luka Bakar Pada Tikus Sprague Dawley Kajian Pada: Tumor Nekrosis Faktor - Alfa dan Perbaikan Luas Luka Bakar

Latar Belakang: Luka bakar merupakan masalah klinis dengan komplikasi yang sering menyebabkan morbiditas dan mortalitas. Omega 3 memiliki efek anti inflamasi dan restriksi kalori memiliki efek anti oksidan yang diduga mampu mempercepat proses penyembuhan pada luka. Penelitian bertujuan untuk mengetahui apakah kombinasi suplementasi Omega 3 dan restriksi kalori dapat membantu dalam proses penyembuhan luka bakar yang dapat ditinjau dari kadar TNF Alfa dan perbaikan luas luka bakar. Metode: Penelitian eksperimental dengan desain “Randomized post test with control group” pada tikus yang di intervensi luka bakar pada punggungnya. Random alokasi dilakukan untuk membagi tikus Sprague Dawley dalam 4 kelompok yaitu kelompok kontrol(K1) dengan diet biasa, kelompok restriksi kalori(P1), kelompok suplementasi omega 3(P2) dan kelompok kombinasi restriksi kalori dengan suplementasi omega 3 (P3). Data dianalisis dan diolah menggunakan uji hipotesis Kruskall Wallis - Post Hoc test Mann-Whitney. Hasil: Penelitian didapatkan perbedaan bermakna yang signifikan kadar TNF Alfa pada kelompok Kontrol dengan kelompok P1, P2, dan P3 dengan p < 0,001. Luas luka bakar didapatkan hasil bermakna signifikan antar kelompok dengan p < 0,001. Post Hoc test kadar TNF alfa dan luas luka bakar menunjukkan tidak ada perbedaan bermakna antara kelompok P2 dengan P3. Kesimpulan: Kombinasi pemberian suplementasi omega 3 dengan restriksi kalori dapat menurunkan kadar TNF Alfa dan memperkecil ukuran luas luka bakar secara bermakna namun tidak untuk restriksi kalori saja Kata Kunci: penyembuhan luka, Omega 3, Kalori Restriksi, TNF Alfa Background: Burns are clinical problem with complications that often cause morbidity and mortality. Omega 3 has an anti-inflammatory effect and calorie restriction has an anti-oxidant effect which is thougth to accelerate the healing process in wounds. The research aims to determine wheteher the combination of Omega 3 supplementation and calorie restriction is expected to help in the process of healing burns that can be viewed from the level of tumor necrosis factor alpha and large burn healing. Methods: This study an experimental study with design of “randomized post test with control group” in rats given burns to their backs. Random allocation was made to divide Sprague Dawley rats into 4 groups: the control group with a normal diet (K1), the calorie restriction group (P1), the omega 3 supplementation group (P2), and the combination of omega 3 with calorie restriction group (P3). Data were analyzed and processed using the Kruskall Wallis - Post Hoc Mann-Whitney hypothesis test. Results: The study found significant differences in levels of TNF Alpha in the control group with groups P1, P2, and P3 with p<0.001. Burn area showed significant results between groups with p<0.001. Post Hoc tests for levels of TNF alpha and burn area showed no significant diffrence between P2 and P3 groups. Conclusion: The combination of omega 3 supplementation with calorie restriction can significantly reduce TNF alpha levels and significantly reduce the area of burns but not for calorie restriction only. Keywords: wound healing, omega 3, calorie restriction, TNF Alpha

The Relationship of Intrauterine Growth Restriction with Placental Pathologic Changes in Newborns

Introduction: Intrauterine growth restriction is a multifaceted problem and is associated with a significant increase in the level of morbidity and perinatal mortality. According to some studies, failure of the placenta is responsible for the most cases of intrauterine growth restriction. The aim of this study was to evaluate the placental pathologic changes in the intrauterine growth restriction (IUGR) samples and compare them with normal cases. Methods: A study population consisted of 60 intrauterine growth restriction neonates and 60 normalized neonates born at Tehran Imam Khomeini Hospital between June 2016 and July 2017. The placenta was weighed, immediately after delivery, and the umbilical cord was separated, then stored in 10% formalin and sent for pathological examination as soon as possible. Data collection was performed according to the following items: the pathologist's report, the results of the infants' examination, and the data in the neonatal cases. Results: The intrauterine growth restriction group showed a high frequency of placenta infarction (P < 0.001), inflammation of the villous (P < 0.001), villous fibrosis (P = 0.044), villous vascularization disorder (P = 0.001), prevalence of chorioamnionitis (P = 0.027), prevalence of Syncytiotrophoblastic knots (P < 0.001) and placental necrosis (P = 0.048) than normal group. However, the mean weight of the placenta (P < 0.001), the length and width of the macroscopic placenta changes was less (P < 0.001). Conclusion: The results of the current study showed that a major part of the macroscopic and histological changes are detectable in the intrauterine growth restriction samples, which are considerably more common than normal, although they are not pathognomonic, but in the future, more accurate results can be obtained from more extensive studies.

Early detection of insulin resistance in children with different types of intrauterine growth restriction

Relevance of the research. Intrauterine growth restriction in children is associated with an increased risk of insulin resistance and non-insulin-dependent diabetes mellitus later in life. The influence of type of intrauterine growth restriction and the mechanisms of insulin resistance are still unknown; the role of catch-up growth in this process is controversial. The aim of the study was to identify insulin resistance in children with different types of intrauterine growth restriction and to analyze the role of catch-up growth in this process. Materials and methods. The research involved 95 newborns, which were divided into groups based on birth weight and length: 60 newborns with intrauterine growth restriction (group I – 31 with asymmetrical intrauterine growth restriction; group II – 29 with symmetrical intrauterine growth restriction) and control group (group III) – 35 newborns without intrauterine growth restriction. Children also were divided into groups according to the presence of catch-up growth to 3 months old. The levels of insulin, insulin-like growth factor-1, growth hormone were measured in cord blood at birth and in blood serum at 3 months old. Glucose levels were measured in serum and insulin resistance index “the homeostasis model assessment of insulin resistance” (HOMA-IR) was calculated in children at 3 months. Results. The children with intrauterine growth restrictioncompared to control group had significantly lower levels of insulin-like growth factor-1 in cord blood. The differences between types of intrauterine growth restrictionhave been observed: children with symmetrical intrauterine growth restriction had higher levels of growth hormone, insulin and HOMA-IR then in asymmetrical one. Correlations between insulin and glucose in children with symmetrical intrauterine growth restriction were absent unlike to asymmetrical intrauterine growth restriction (+0.61). The negative role of catch-up growth in insulin resistance development has been defined: it was related to hyperinsulinemia and increased the frequency of insulin resistance in children either in asymmetrical or in symmetrical intrauterine growth restriction, but more in symmetrical one. Conclusion. Children with symmetrical intrauterine growth restriction especially with catch-up growth are at the highest risk of metabolic syndrome development in later life and require increased monitoring by pediatricians and endocrinologists.

Intradialytic exercise with blood flow restriction is more effective than conventional exercise in improving walking endurance in hemodialysis patients: a randomized controlled trial

Objective: This study aims to compare the effect of intradialytic aerobic exercise with blood flow restriction, without blood flow restriction (conventional) and no exercise (control) on muscle strength and walking endurance among chronic kidney disease patients. Design: Open label and parallel group randomized controlled trial. Subjects: Adult patients with chronic kidney disease on hemodialysis. Intervention: A 12-week intradialytic training with or without blood flow restriction compared with a control group. Main measures: Strength and walking endurance were measured using thoracolumbar dynamometry and a 6-minute walk test, respectively, before and after training. Results: A total of 66 patients were randomized into three groups: blood flow restriction group ( n = 22), conventional exercise group ( n = 22) and control group ( n = 22). There were seven dropouts, and 59 patients were included in the analysis. There was a significant increase in the 6-minute walking distance in the blood flow restriction group (from 412.7 (115.9) to 483.0 (131.0) m, P = 0.007) in comparison with the conventional exercise group (from 426.79 (115.00) to 433.2 (120.42) m, not significant) and the control group (from 428.4 (108.1) to 417.3 (100.2) m, not significant). The change in the walking distance over time was significantly different among groups (intervention group/time, P = 0.02). The simple effects test found a significant time effect only in the blood flow restriction group. There was no significant difference in strength change between the groups. Conclusion: Among chronic kidney disease patients, intradialytic exercise of low/moderate intensity with blood flow restriction was more effective in improving walking endurance than conventional exercise or no exercise.

175 THE EFFECT OF NUTRITION ON OVARIAN FOLLICLE POPULATION AND PLASMA ANTI-MULLERIAN HORMONE CONCENTRATION IN ABERDEEN ANGUS HEIFERS

The number of follicles in ovaries from cows is highly variable and has a high repeatability in females during the follicular waves. There is evidence that dietary restriction during pregnancy has a deleterious effect on fetal ovary reserve (Mossa et al. 2009). The anti-Mullerian hormone (AMH) is a member of the TGFβ superfamily, produced by granulosa cells from healthy growing follicles, and its concentration is correlated with the number of follicles recruited in the wave during the oestrous cycle and may be used as a biomarker of the follicle population in cattle. Our objective was to compare the number of follicles and AMH concentration in heifers born from cows that suffered food restriction during the first trimester of pregnancy (March to May of 2009) to heifers born from cows that did not suffer food restriction (March to May of 2010) in Bagé, RS, Brazil. Because the diet of these cows were pasture based when rain dropped from an average of 129 mm3 (from 2002 to 2008) to 64 mm3 (in 2009), these cows suffered food restriction. In 2010 rain averaged 83 mm3, reestablishing normal feeding conditions. Ovarian follicle count analysis and blood drawing was performed in 89 heifers born in 2009 and 131 heifers born in 2010. The counting of antral follicles (>3 mm) by ultrasound was only in heifers with absence of follicles >6 mm at the time of assessment. Cows from both years were also grouped into low (LFC) or high (HFC) follicle count. Plasma AMH concentrations (pg mL–1) were determined by ELISA using human AMH/MIS Gen II commercial kit (Beckman Coulter, Brea, CA, USA) validated using plasma from cows of HFC and LFC, a castrated bull, and a pool of follicular fluid. Results were analysed using the Proc GLM procedure of SAS (SAS 9.2). Follicle LSmean was higher (P < 0.05) in heifers that did not suffer food restriction (19.09 ± 0.8) when compared with heifers born from cows that suffered food restriction (11.75 ± 0.75). Within no food restriction and food restriction groups, LFC heifers averaged 10.91 ± 1.21 and 4.16 ± 1.02 follicles, respectively, and HFC heifers averaged 27.28 ± 1.0 and 19.35 ± 1.0 follicles, respectively. Anti-Mullerian hormone concentration was higher (P < 0.05) for heifers born in 2010 (no food restriction; 77.24 ± 4.71 pg mL–1) when compared with heifers born in 2009 that mothers suffered food restriction (30.89 ± 4.32 pg mL–1). Furthermore, within heifers from the food restriction group AMH concentration was higher (P < 0.05) in HFC heifers (47.17 ± 6.2) when compared with LFC heifers (14.61 ± 6.0). However, there was no difference in AMH concentration between LFC and HFC in the no-food restriction group. In conclusion, dietary restriction during the first trimester of pregnancy might result in reduced follicle population in the female offspring; however, caution should be taken since animal variation is consistent. Furthermore, AMH concentration can be used as a biomarker for follicle population. This research and scholarship for Ereno, Pupulim, Loureiro, Favoureto, and Castilho is from FAPESP.

Evaluation of foliar fungal endophyte incidence in field-grown transgenic Bt white spruce trees

A total of 770 transgenic Bt white spruce needles were collected and plated on potato dextrose agar to determine their foliar endophyte diversity. The ribosomal internal transcribed spacer regions for 310 foliar endophytes were amplified by polymerase chain reaction (PCR) and digested using CfoI and MspI, which created 21 restriction groups. Isolates from each restriction group were sequenced and compared with reference sequences in GenBank. Eighteen sequence groups were obtained, of which five were identified at the species level. The most common endophytic fungi identified by PCR-RFLP was Lophodermium piceae (incidence of 74.5%). The second and third most common ones were Hypoxylon fragiforme (3.63%) and Lophodermium nitens (3.18%). A statistical analysis performed on the most common endophyte groups showed no statistical difference in endophyte frequency or distribution between the control white spruce needles (nontransgenic) and saplings with constructs containing the reporter gene GUS or the Bt Cry1A(b) gene and kanamycin.

Changes in Levels of Serum Insulin, C-Peptide and Glucose after Electroacupuncture and Diet Therapy in Obese Women

Our purpose was to investigate the effects of electroacupuncture (EA) therapy on body weight and on levels of serum insulin, c-peptide and glucose in obese women. 52 healthy women were included in this study and were allocated into three groups: 1) Placebo EA group ( n = 15; mean age = 41.8 ± 4.6 and mean body mass index { BMI } = 33.2 ± 3.5); 2) EA group ( n = 20; mean age = 42.1 ± 4.4 and BMI = 35.9 ± 3.6) and 3) Diet restriction group ( n = 20; mean age = 42.9 ± 4.3 and BMI = 34.7 ± 2.7). EA was applied to the ear points Hunger and Shen Men on alternating days and to the body points LI 4, LI 11, St 36 and St 44 once a day for 30 minutes over 20 days. Diet restriction that entailed a 1450 kilocalorie (kcal) diet program was applied to the three groups for 20 days. An increase in weight loss was observed when weight loss in the EA group (p < 0.000) was compared to that in the diet restricted and placebo EA groups using the Tukey HSD test. There were increases in the serum insulin (p < 0.001) and c-peptide levels (p < 0.000) in the women treated with EA compared to those in the women treated with the placebo EA and diet restriction groups. A decrease was observed in the glucose levels (p < 0.01)in both the EA and diet restriction groups compared to those in the placebo EA group. Our results suggest that EA therapy is an effective method in treating obesity. EA therapy also helps serum glucose levels to decrease through the increase of serum insulin and c-peptide levels.

Molecular characterization of Coxiella burnetii isolates

Restriction fragment length polymorphism (RFLP) was used for the differentiation of 80 Coxiella burnetii isolates derived from animals and humans in Europe, USA, Africa and Asia. After NotI restriction of total C. burnetii DNA and pulsed field gel electrophoresis (PFGE) 20 different restriction patterns were distinguished. The index of discrimination for this typing system was 0·86. Comparison and phylogenetic analysis of the different RFLP patterns revealed evolutionary relationships among groups that corresponded to the geographical origin of the isolates. This finding was confirmed by genetic mapping. No correlation between restriction group and virulence of isolates was detected.