Distribution and immunohistochemical characterisation of paracervical neurons innervating the oviduct in the pig

The present study was aimed at disclosing the distribution of paracervical neurons projecting to the ampulla and isthmus of the porcine oviduct and the pattern(s) of co-existence of tyrosine hydroxylase (TH), dopamine ß-hydroxylase (DßH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP) and nitric oxide synthase (NOS) within these nerve cell bodies. The fluorescent retrograde tracer Fast Blue (FB) was injected into the wall of the ampullar (n = 3) and isthmal (n = 3) part of the organ in six sexually immature female pigs. After a survival period of three weeks paracervical ganglia (PCG) were collected. 10 µm-thick cryostat sections of the ganglia were examined for the presence of FB-positive (FB+) nerve cells under the fluorescent microscope. Tracered neurons were counted in every third section and processed for double-labelling immunofluorescence according to the method of Wessendorf and Elde. 78.6% of FB+ neurons were projecting to the isthmus while 21.4% of the studied population innervated the ampulla of the oviduct. Double-labelling immunofluorescence revealed the existence of the following different chemically coded subpopulations of the studied perikarya: TH+/D bH+, TH+/NPY+, TH+/NOS+, TH+/ NOS-, SP-/NOS+, SP+/CGRP+

A high manganese-tolerant pseudomonas sp. strain isolated from metallurgical waste heap can be a tool for enhancing manganese removal from contaminated soil

Manganese (Mn) is widely used in industry. However, its extensive applications have generated a great amount of manganese waste, which has become an ecological problem and has led to a decrease in natural resources. The use of microorganisms capable of accumulating Mn ions from contaminated ecosystems o ers a potential alternative for the removal and recovery of this metal. The main aim of this work was an investigation of removal potential of Mn from soil by isolated bacterial. For this purpose, eleven bacterial strains were isolated from the soil from metallurgical waste heap in Upper Silesia, Poland. Strain named 2De with the highest Mn removal potential was selected and characterized taking into account its ability for Mn sorption and bioaccumulation from soil and medium containing manganese dioxide. Moreover, the protein profile of 2De strain before and after exposition to Mn was analyzed using SDS/PAGE technique. The 2De strain was identified as a Pseudomonas sp. The results revealed that this strain has an ability to grow at high Mn concentration and possesses an enhanced ability to remove it from the solution enriched with the soil or manganese dioxide via a biosorption mechanism. Moreover, changes in cellular protein expression of the isolated strain were observed. This study demonstrated that autochthonous 2De strain can be an e ective tool to remove and recover Mn from contaminated soil

RHD variant in RhD/-/ mother with anti-D makes noninvasive fetal RHD genotyping impossible

Abstract Objectives: Noninvasive fetal RHD genotyping from maternal plasma of RhD(/-) pregnant women of Caucasian race may be used for predicting the risk of hemolytic disease because the RHD gene is usually absent in such populations. If detected in plasma of such women, the RHD gene originates from the RhD(+) fetus. The number of fetal copies of the gene in maternal plasma is extremely small. In the presented case of the RhD(/-) pregnant woman with anti-D it was impossible to give a fetal RHD result due to mother’s RHD(+) genotype. The fetal RHD was determined from amniocytes. Aim: to present the difficulties related to the interpretation of results of invasive and noninvasive procedures. Material and methods: whole blood, plasma and amniotic fluid of the RhD(-) woman with anti-D (14 week of pregnancy) as well as whole blood of the newborn. RHD and RHCE*c were genotyped by real-time PCR in DNA isolated from maternal plasma and amniocytes and the RHD and d-genotypes were tested by SSP methods in DNA isolated from whole blood and amniocytes . Results: RHD and RHCE*c were detected in DNA isolated from plasma. The high level of RHD suggested its origin from the mother’s DNA therefore it was impossible to determine the fetal RHD. The d-little test identified a RHD(IVS3+1G>A) variant in the mother’s genome. A weak signal of real-time PCR for the RHD was obtained in amniocytes but the RHD was not detected by SSP. The RHCE*c was detected by both methods. Results were inconclusive; the fetal RHD status remained unknown. The child was RhD(-) with RHD in its DNA undetected by either method. Conclusions: 1/ The RHD(IVS3+1G>A) variant in the RhD(-) mother precluded formal noninvasive fetal RHD genotyping. 2/Real-time PCR is too sensitive for amniocyte testing and may lead to false results as it detects trace maternal DNA in amniotic fluid 3/ The frequency of RHD(IVS3+1G>A) occurrence in Poland requires further studies

Alloimmunizacja u chorych na niedokrwistość autoimmunohemolityczną oraz genotypowanie krwinek czerwonych w celu udoskonalenia doboru krwi do przetoczeń

Wstęp: Autoprzeciwciała typu ciepłego występujące w surowicy chorych na niedokrwistość autoimmunohemolityczną powodują niezgodność w próbie zgodności i utrudniają wykrycie klinicznie istotnych alloprzeciwciał, stwarzając ryzyko hemolitycznego powikłania poprzetoczeniowego. Dobór krwi do przetoczenia dla tych chorych wymaga szczególnych procedur dla większego bezpieczeństwa transfuzji. Materiał i metody: Analizowano częstość alloimmunizacji u 163 chorych z NAIH (155 z autoprzeciwciałami typu ciepłego, 8 typu mieszanego). U wszystkich chorych określano fenotyp w antygenach Rh, K, konieczny dla doboru krwi do przetoczenia, a u 53 spośród nich, badano ponadto fenotyp Kidd, Duffy, S, s, k jak również stosowano metodę genotypowania krwinek czerwonych. Wyniki: Alloprzeciwciała istotne klinicznie wykryto u 31 chorych (19%). Łącznie zidentyfikowano 42 przeciwciała o różnej swoistości (u 7 chorych występowały przeciwciała o dwóch lub więcej swoistościach) z układów grupowych: Rh, Kell, Kidd, Duffy, MNS i LW. U 14 chorych z alloprzeciwciałami, wykrycie ich wymagało uprzedniego wyadsorbowania autoprzeciwciał z surowicy metodą auto- lub alloadsorpcji. W grupie 53 chorych, fenotyp Rh, K, Kidd i S udało się określić u 37 (70%) w testach serologicznych, zaś fenotyp rozszerzony o antygeny Duffy, s i k, tylko u 9 chorych (17%). Wyniki fenotypowania były zgodne z oznaczeniami genetycznymi. U pozostałych chorych o rozszerzonym fenotypie wnioskowano na podstawie genotypowania, ponieważ wyniki serologiczne nie były miarodajne - dwie populacje krwinek (przetoczenia krwi w ostatnich 3 miesiącach) lub/i autoaglutynacja krwinek chorego. Wnioski: Wprowadzenie metod genotypowania obok metod fenotypowania okazało się konieczne dla ustalenia rozszerzonego fenotypu krwinek czerwonych u chorych z NAIH. Badania te usprawniają dobór krwi do przetaczania i zwiększają bezpieczeństwo transfuzji.Background: The presence of warm autoantibodies in the sera of patients with autoimmune haemolytic anaemia are the cause of incompatibility in the compatibility test, which produces difficulties in the detection of clinically significant alloantibodies thus generating the risk of haemolytic transfusion reactions. For such patients the selection of red cells for transfusion requires special procedures to ensure safety. Material and methods:Alloimmunization was analyzed in 163 AIHA patients (155 with warm type and 8 with mixed type of autoantibodies). In all patients, red cell phenotyping for Rh, K, was performed prior to transfusion and in 53 of them, red cells were phenotyped additionally in Kidd, Duffy, S, s, k with genotyping as well. Results: Clinically significant alloantibodies were found in 31 patients (19%). In all, 42 antibodies of different specificities were identified (in 7 cases we detected antibodies with two or more specificities) within the following blood group systems: Rh, Kell, Kidd, Duffy, MNS and LW. In 14 of the alloimmunized patients, auto- or alloadsorption was required for alloantibody detection. In the group of 53 patients, phenotyping of Rh, K, Kidd and S antigens was successful in 37 patients (70%) with serological tests only, whereas the extended phenotype for Duffy, s, k antigens was successful only in 9 patients (17%). In each case, the genotype results were consistent with the phenotype. In the remaining patients, the results of extended phenotyping with serological method proved unreliable, due either to mixed-field reactions (after recent transfusion < 3 months) or autoagglutination. Genotyping was useful for predicting the phenotype in such cases. Conclusions: In addition to phenotyping, genotyping of red cells is necessary for establishing the extended phenotype in AIHA patients. Such procedure is important for blood transfusion safety