Proteomic analysis of pikeperch seminal plasma provides novel insight into the testicular development of domesticated fish stocks.

Control of the reproduction of domesticated stocks is considered a prerequisite for aquaculture development of pikeperch. However, knowledge about the physiology of the captive pikeperch male reproductive system and the biology of semen is very limited, especially regarding protein characteristics. The aims of our study were to characterize pikeperch sperm quantity and quality parameters and to analyze changes in the proteome of the same males spawned for the first and second times. Moreover, attempts were made to generate the first proteomic library of seminal plasma proteins. Semen collected during the first spawning season was characterized by lower sperm concentration and volume than for the second season. Using mass spectrometry-based label-free quantitative proteomics, we identified 850 proteins in the seminal plasma of pikeperch from both spawning seasons, and 65 seminal proteins were found to be differentially abundant between the first and second spawning seasons. The majority of differentially abundant proteins were involved in stress and immune responses, developmental processes, cofactor metabolic processes, proteolysis, cellular oxidant detoxification and organization of the extracellular matrix (ECM). In addition, several proteins unique to pikeperch seminal plasma were identified, including antifreeze proteins, hibernation-specific plasma proteins, lectins and vitellogenin. In summary, our results indicate that males that spawned for the first time were characterized by incompletely mature gonads and the expression of proteins associated with the early phase of spermatogenesis and ECM organization. On the other hand, males that spawned for the second time exhibited advanced gonadal maturation and expression of proteins related to the late stage of spermatogenesis and sperm maturation, including regulation of reactive oxygen species generation, bicarbonate production, sperm elongation and separation. The identification of a large number of seminal plasma proteins provides a valuable resource for understanding the functions of seminal plasma and the molecular mechanisms involved in testicular development and maturation in domesticated fish, which is a prerequisite for better control of reproduction in captivity.

Caloric restriction can affect one-carbon metabolism during pregnancy in the rat: A transgenerational model.

One-carbon metabolism is critical to pregnancy outcomes, because it determines the availability of nutrients involved in cell divisions and DNA methylation. The aim of this study was to analyze how 50% prenatal calorie restriction affected one-carbon metabolism in pregnant Wistar rats of the F0 to F2 generations. Mean choline (p < 0.001), betaine (p < 0.001), and S-adenosylmethionine (SAM) (p < 0.05) concentrations were respectively about 40%, 45%, and 20% lower in the F0_R (R - restricted diet) than in the F0_C (C - control diet). Homocysteine, S-adenosylhomocysteine (SAH), and trimethylamine oxide concentrations were unaffected. In the F1_R, the SAM-to-SAH ratio was 25% higher (p < 0.05) than in the F1_C. No differences between the C and R groups were observed in the F2 generation. The SAM concentrations in the F1_R were higher than in the F0_R and the F2_R (p < 0.01). The relative transcript levels of Mat1a, Bhmt, Cbs, Pemt, and Mthfr were only slightly affected by the diet, with changes of less than a factor of 2.0. Cbs activity in the F2_R was significantly higher than in the F2_C (p < 0.001). Food deprivation may affect one-carbon metabolism in pregnant rats, but it does not stimulate persistent metabolic changes that can be observed during the pregnancy of their progeny of the F1 or F2 generations.

How to switch the motor on: RNA polymerase initiation steps at the single-molecule level.

RNA polymerase (RNAP) is the central motor of gene expression since it governs the process of transcription. In prokaryotes, this holoenzyme is formed by the RNAP core and a sigma factor. After approaching and binding the specific promoter site on the DNA, the holoenzyme-promoter complex undergoes several conformational transitions that allow unwinding and opening of the DNA duplex. Once the first DNA basepairs (∼10 bp) are transcribed in an initial transcription process, the enzyme unbinds from the promoter and proceeds downstream along the DNA while continuously opening the helix and polymerizing the ribonucleotides in correspondence with the template DNA sequence. When the gene is transcribed into RNA, the process generally is terminated and RNAP unbinds from the DNA. The first step of transcription-initiation, is considered the rate-limiting step of the entire process. This review focuses on the single-molecule studies that try to reveal the key steps in the initiation phase of bacterial transcription. Such single-molecule studies have, for example, allowed real-time observations of the RNAP target search mechanism, a mechanism still under debate. Moreover, single-molecule studies using Förster Resonance Energy Transfer (FRET) revealed the conformational changes that the enzyme undergoes during initiation. Force-based techniques such as scanning force microscopy and magnetic tweezers allowed quantification of the energy that drives the RNAP translocation along DNA and its dynamics. In addition to these in vitro experiments, single particle tracking in vivo has provided a direct quantification of the relative populations in each phase of transcription and their locations within the cell.

Comparison of serum concentrations of environmental allergen-specific IgE in atopic and healthy (nonatopic) horses.

Allergic responses in humans, horses and other species are mediated by immunoglobulin E (IgE) antibodies. Serum testing to detect allergen-specific IgE antibodies has been developed for dogs, cats and horses; this allows for the identification of allergens and determination of appropriate allergen- specific immunotherapies. This study compared serum allergen-specific IgE concentrations in atopic and healthy horses. The study was performed on Malopolski breed atopic (n=21) and nonatopic (n=21) clinically healthy horses. Allergen-specific IgE serum concentrations were measured in summer seasons of 2008-2015 using a monoclonal anti-IgE antibody. A Northern and Central European allergen panel containing mite, insect, mould and plant pollen allergens, including 15 tests of individual allergens and 5 tests of allergen mixtures was used. The mean allergen-specific IgE concentrations in the atopic and normal horse populations were compared. Among the atopic horses, the strongest positive reactions occurred against the storage mites Tyrophagus putrescentiae and the domestic mite Dermatophagoides farinae. The atopic horses also demonstrated high IgE concentrations against insects, particularly Tabanus sp., the plant pollens colza, cultivated rye and the mould pollen mixture Aspergillus/Penicillium. No horses in the atopic group were IgE-negative. Among all mite, insect, mould and some plant allergen groups the differences in mean specific IgE concentrations between allergic and healthy horses were significant. The mean IgE concentrations for most allergen groups were significantly higher in the atopic horses than in the healthy animals. However, a high incidence of positive reactions was observed in both healthy and allergic horses. Our results showed a high frequency of polysensitization in atopic horses.

Synthesis and cellular activity of stereochemically-pure 2'-O-(2-methoxyethyl)-phosphorothioate oligonucleotides.

Stereochemically-pure 2'-O-(2-methoxyethyl)-phosphorothioate (PS-MOE) oligonucleotides were synthesized from new chiral oxazaphospholidine-containing nucleosides. Thermal stability studies showed that the incorporation of Rp-PS linkages increased RNA-binding affinity. In cells, a full Rp-PS-MOE splice-switching oligonucleotide targeting part of the ferrochelatase gene was more potent than its Sp-PS counterpart, but of similar potency to the stereorandom PS-parent sequence.

Characterization of solid state nuclear track detectors of the polyallyl-diglycol-carbonate (CR-39/PM-355) type for light charged particle spectroscopy.

This paper presents a method which uses the characteristics of the etch pits induced in a polyallyl-diglycol-carbonate (PADC) detector of the CR-39/PM-355 type to estimate particle energy. This method is based on the data provided by a semiautomatic system that selects tracks according to two parameters, crater diameters, and mean gray level values. In this paper we used the results of the calibration measurements that were obtained in our laboratory in the period 2000-2014. Combining the information on the two parameters it is possible to determine unambiguously the incident projectile energy values. The paper presents the results of an attempt to estimate the energy resolution of the method when analyzing the tracks produced in the CR-39/PM-355 detector by energetic ions such as alpha particles, protons, and deuterons. We discuss the energy resolution of the measurement of light charged particle energy which is based on the parameters (crater diameter and mean gray level value) of tracks induced in solid state nuclear track detectors of the PADC type.

Investigations of protons passing through the CR-39/PM-355 type of solid state nuclear track detectors.

Solid State Nuclear Track Detectors of the CR-39∕PM-355 type were irradiated with protons with energies in the range from 0.2 to 8.5 MeV. Their intensities and energies were controlled by a Si surface barrier detector located in an accelerator scattering chamber. The ranges of protons with energies of 6-7 MeV were comparable to the thickness of the PM-355 track detectors. Latent tracks in the polymeric detectors were chemically etched under standard conditions to develop the tracks. Standard optical microscope and scanning electron microscopy techniques were used for surface morphology characterization.

Influence of high temperature on solid state nuclear track detector parameters.

This work concerns the influence of high temperatures on tracks induced in solid state nuclear track detectors of the CR-39/PM-355 type. In order to investigate this effect some samples of the detectors were irradiated with energetic protons and α particles and subsequently heated under controlled temperatures for different periods of time. After heating the samples were etched and the track evolution was analyzed using an optical microscope. The bulk etch rate V(B) of the PM-355 material was also determined as a function of heating temperature. The track etch rate V(T) values were estimated for craters induced by protons and α particles from track diameter measurement as a function of heating temperature.

Evaluation of CXCL10, CXCL11, CXCL12 and CXCL13 chemokines in serum and cerebrospinal fluid in patients with tick borne encephalitis (TBE).

PURPOSE: The aim of the study was to assess the concentration of chemokines: CXCL10, XCL11, CXCL12, CXCL13 in serum and cerebrospinal fluid (CSF) in patients with tick-borne encephalitis (TBE) before and after treatment. We evaluated also the usefulness of these molecules in diagnosis and monitoring of inflammation in TBE. METHODS: Twenty three patients hospitalized in The Department of Infectious Diseases and Neuroinfections of Medical University in Bialystok, Poland were included in the study. Patients were divided into 2 groups: TBE group-patients with confirmed TBE and control group (CG): patients with excluded TBE and other inflammatory diseases of CNS. Concentration of CXCL10/IP-10, CXCL11/I-TAC, CXCL12/SDF-1α, CXCL13/BLC/BCA-1 in serum and CSF were measured with ELISA kits (R&D Systems, USA) according to the protocols. RESULTS: The analysis of chemokines concentration in TBE patients before treatment and control group using ROC showed that serum CXCL10 and CXCL13 and CSF CXCL10, CXCL11, CXCL12 and CXCL13 differentiate both groups (p<0.05). The analysis of CXCL10, CXCL11, CXCL12 and CXCL13 before and after treatment showed that CXCL10 and CXCL11 in CSF and CXCL13 in serum differentiates both groups with p<0.05. CONCLUSIONS: Concentration of CSF CXCL10, CXCL11, CXCL12, CXCL13 and serum CXCL10, CXCL13 may be good biomarkers of CNS inflammation caused by TBEV. Moreover concentration of CXCL10 in CSF and CXCL13 in serum may be used as indicators of patients recovery.

Identification of proteins co-purifying with scrapie infectivity.

PrP(C), the cellular isoform of prion protein, is widely expressed in most tissues. Despite its involvement in several bioprocesses it still has no apparent physiological role. During propagation of Transmissible Spongiform Encephalopathies, PrP(C) is converted to the pathological isoform, PrP(Sc), in a process believed to be mediated by unknown host factors. PrP(Sc) has altered biochemical properties and forms amyloid aggregates that display infectious characteristics. PrP(Sc) is also the major component in biochemically enriched infectious samples. Other molecules co-purify with it, but the protein content of these aggregates remains unknown. The goal of this project was to identify other host molecules with high affinity for PrP(Sc). Here, we present the identification of protein molecules that co-purify with PrP(Sc) isolated from naturally scrapie-infected ovine brain tissue. The infectious preparations were analyzed by two-dimensional gel electrophoresis and unknown proteins were identified by LC-MS/MS. These proteins may prove to be strategic targets for prevention and therapy of prion diseases.

[Results of screening tests for gestational diabetes mellitus at Medical University in Warsaw]. / Badania przesiewowe w kierunku rozpoznawania cukrzycy ciezarnych na materiale I i II Kliniki Poloznictwa i Ginekologii Akademii Medycznej w Warszawie.

Our purpose was to determine the incidence of screening for gestational diabetes among the population of women delivering at I and II Departments of the First Faculty of Medical University in Warsaw. A retrospective review of 647 pregnancies was performed. The incidence of gestational diabetes mellitus screening was determined and the rate of occurrence of GDM analyzed. 310 (48%) pregnancies were screened for gestational diabetes mellitus with a 1-hour, 50 gm oral glucose challenge test. 49 (16.07%) of the screens had positive results at a plasma glucose level of > 139 mg/dl. Two-hour 75 gm oral glucose tolerance tests (according to the 1994 World Health Organization panel recommendations) were performed on screen-positive women, eleven of whom (22.45%) were diagnosed with gestational diabetes mellitus. Despite of positive oral 50 gm glucose test, (plasma glucose level 140-179 mg/l) 15 women (30%) haven't had the 75 gm oral glucose test. The incidence of GDM among analyzed population is 4% and when GDM screening is carried out, exceeds 7%. Early gestational glucose screening, if performed, may be beneficial in detecting gestational diabetes. Consideration should be given to fulfill it more frequently and for sure, repeat glucose testing in patients with positive one-hour screening tests.

[Risk factors for gestational diabetes mellitus and results of glucose tolerance tests]. / Czynniki ryzyka cukrzycy ciazowej a wyniki testów obciazenia glukoza.

The authors made an effort to verify the connection between the presence of risk factors for GDM and results of screening and diagnostic tests. Study group consisted 302 patients. Gestational diabetes was more frequently diagnosed when an excessive maternal weight and family history of diabetes occurred. Among women with gestational diabetes recognised on the basis of the tests results (screening or diagnostic), 1/3 of patients had no evidence of any risk factor. There is no correlation between the number of risk factors and the occurrence of gestational diabetes. The risk factors were present in half of the investigated patients.

Lichtenstein inguinal hernia repair in a primary healthcare setting.

BACKGROUND: Given the generally accepted poor outcome of inguinal hernia repair when using nylon darn, and the recent interest in low-tension mesh repair, an attempt was made to demonstrate the feasibility, outcome and patient perception of providing Lichtenstein inguinal hernia repair, using local anaesthesia, wholly within the primary healthcare sector. METHODS: A prospective study reviewed clinical outcome and patient perception in 100 adults referred with inguinal hernia only. No selection was made regarding age, sex, American Society of Anesthesiologists status or previous repairs. Recurrence, pain, infection, return to full function and associated complications were assessed at 24 h, 1 and 6 weeks, and 1 year. Local Community Health Councils assessed patient perception. RESULTS: In the first 100 patients (age range 21-83 (mean(s.d.) 60(14.7)) years; 58 of employable age; 92 men; ten recurrent hernias), no recurrences have occurred at 1 year. Infection rate was 3 per cent. Pain was maximal in the first 24-48 h (median visual analogue scale 5, range 0-10) and reduced rapidly (median 1) at 1 week. Mean time to return to work or full normal activity was 8 days. Some 85 operations were performed within 1 month of diagnosis. In all, 86 patients returned the patient satisfaction questionnaire and 98 per cent of these were 'very pleased' with the service. CONCLUSION: In highly motivated primary healthcare centres, inguinal hernia repair can be undertaken effectively, providing high patient satisfaction, minimal complications and low recurrence rates using the Lichtenstein technique.

[P1 blood group, secretion status and susceptibility to asymptomatic bacteriuria in diabetes]. / Grupa krwi P1 oraz stan wydzielania a predyspozycja do bakteriurii bezobjawowej w cukrzycy.

Blood-group antigens found on uroepithelial cells and in the secretions may affect bacterial adherence and thereby the predisposition to urinary tract infection. We determined P1, Lewis-blood-group phenotype and secretor status in patients with diabetes mellitus: 12 with asymptomatic bacteriuria and 7 without its presence. There was no difference between the two groups in the distribution of the P1 phenotype. There was also no statistical difference in the distribution of the Lewis phenotype and secretor status, although there appeared to be general trend of higher number of Le (a+b-) phenotype and non-secretors present in the asymptomatic bacteriuria group. Further studies are necessary to determine the role of blood groups and secretor status in the pathogenesis and susceptibility to urinary tract infection.

Distribution of iron in pig placenta (Sus scrofa dom. L.).

Iron distribution was studied in pig placentae between 21 day till the end of pregnancy (113) with the use of histochemical and cytochemical methods and X-ray microanalysis. Iron content was measured in fetal and maternal part of the placenta with chemical methods. Iron presence was confirmed in maternal and fetal erythrocytes, cells and secretion of uterine glands and trace amount in trophoblast lining regular areolae. No significant differences were found in iron content in fetal and maternal part of the placenta throughout the entire studied period. With the applied histochemical method of iron determination according to Perls, potassium ferrocyanide also adsorbs in sites where mucopolysaccharides are present, in which iron presence has not been detected with the use of X-ray microanalysis.

[Effect of the placental barrier on iron levels in biological fluids and tissues of sows and their fetuses during pregnancy]. / Wplyw bariery lozyskowej na zawartosc zelaza w plynach biologicznych i tkankach macior i ich plodów w przebiegu ciazy.

The investigation was carried out on 4 gilts and 29 pregnant sows (in the pregnancy period from 21st to 112th day) as well as their fetuses. The sows and their fetuses were divided into 4 groups with regard to the duration of pregnancy. Total iron binding capacity (TIBC) and unbound iron binding capacity (UIBC) were determined and percent of transferrin saturation was calculated in the blood serum of the sows and in the serum of umbilical blood as well as in amniotic and allantoic liquids. Iron centent in the uterus, placenta, liver, kidneys and spleen was determined by the method of atom absorption. The results were converted to 1 kg of dry matter of the organ. It was found that transferrin was not the only transporting form of iron in the fetuses of sows, though it was predominant in the majority of the examined fetuses. It can be assumed that other transporting form depends mainly on the saw itself and participates in Fe flow through placenta. It is still unknown if transferrin being present in amniotic and allantoic liquid explicitly originates from fetus. The organs revealing the highest contribution to iron mobilization during pregnancy seem to be spleen and then liver.

[Dynamics of quantitative changes in mineral components of body fluids and organs of sows and their fetuses in ontogenic development]. / Dynamika zmian ilosciowych skladników mineralnych plynów ustrojowych i narzadów macior oraz ich plodów w rozwoju ontogenetycznym.

The experiment was carried out on 4 gilts and 29 pregnant sows and their fetuses (pregnancy period from 21st to 112th day). The animals were divided into groups in relation to the duration of pregnancy. The content of Na, K, Mg and Ca in the serum of sows and fetuses and also in several organs of the sows and fetuses was determined by the method of atomic absorbtion whereas the content of inorganic and total P was determined by colorimetric method. The results were converted into 1 kg of dry matter content. It was found that there is difference in administration of Ca and Na during pregnancy of swine in compare with other species. During pregnancy large amounts of these elements accumulate in placenta but their increase in the serum of fetuses becomes unaffected, although this occurs in other species. Inner organs of sows, i.e. liver, kidneys and spleen accumulate considerable amounts of Na and Ca, especially in early pregnancy, towards the the end of pregnancy the content of all mineral components in the liver of fetus decreases, whereas in kidneys only the content of Na and Ca, and in the spleen only Na content is depressed. In the fetuses liquids several irregular changes in the concentration of mineral substances were found, particularly in the allantoic fluid.

The effect of malathion on RNA polymerase activity of cell nuclei and transcription products in lymphocyte culture.

Malathion decreased polymerase activity of cell nuclei and inhibited RNA synthesis in cultured lymphocytes. The mechanism of action of this compound at the cellular level is suggested.