Kinetics of murine decidual dendritic cells

Dendritic cells (DCs) are professional antigen presenting cells (APC) capable of induction of primary immune responses as well as immunologic tolerance. Myeloid and lymphoid subsets of murine DCs are able to shift cytokine responses of T cells toward Th2 and Th1 profiles respectively. Thus, DCs would be suitable candidates to mediate the balance of maternal immune responses to conception. We analyzed pregnancy-related variations in uterus and splenic DCs in a murine model. C57BL/6-mated Balb/c female mice with vaginal plugs were scarified at early, middle, and late pregnancy. Frozen sections of uterus and spleen at each stage of pregnancy were immunostained with CD11c- and MHC-II-specific antibodies. Two-color immunohistochemistry was also carried out using anti-CD11c and one of the antibodies against CD11b, CD8α, CD86, and DEC-205. Using morphometric analysis, the average density of DCs and relative percentage of myeloid (CD11c+, CD11b+) and lymphoid DCs (CD11c+, CD8α+) were determined at each stage. Our results showed that DCs are present throughout the pregnancy in decidua. The average density of decidual DCs at early pregnancy was significantly higher relative to middle and late gestation or to those of endometrial DCs of non-pregnant mice. Interestingly, the average density of decidual and splenic DCs, followed the same variations at different stages of pregnancy. The relative percentage of decidual lymphoid DCs (LDC) was significantly higher at mid-gestation when compared with other stages of pregnancy or non-pregnant mice. Inversely, the frequency of myeloid DCs (MDC) and the MDC/LDC ratio were statistically lower at the middle stage of pregnancy. A majority of decidual DCs expressed MHC-II and CD86. At early pregnancy, DCs were more concentrated subadjacent to the luminal epithelial layers, whereas at mid- or late gestation, DCs were randomly distributed in the stroma and around the epithelium. Mid-pregnancy period was a critical point with regard to splenic DCs kinetics, as both the average density of DCs and the frequency of MDCs decreased significantly when compared with early or late pregnancy, although the relative percentage of splenic LDCs did not change. Our data suggest that the balance of MDC and LDC is finely tuned throughout pregnancy, pointing an eminent immunoregulatory role of DCs in the maintenance of pregnacy. © 2007 Society for Reproduction and Fertility

BVVL/ FL: features caused by SLC52A3 mutations; WDFY4 and TNFSF13B may be novel causative genes

Brown-Vialetto-Van Laere (BVVL) and Fazio-Londe are disorders with amyotrophic lateral sclerosis-like features, usually with recessive inheritance. We aimed to identify causative mutations in 10 probands. Neurological examinations, genetic analysis, audiometry, magnetic resonance imaging, biochemical and immunological testings, and/or muscle histopathology were performed. Mutations in known causative gene SLC52A3 were found in 7 probands. More importantly, only 1 mutated allele was observed in several patients, and variable expressivity and incomplete penetrance were clearly noted. Environmental insults may contribute to variable presentations. Putative causative mutations in other genes were identified in 3 probands. Two of the genes, WDFY4 and TNFSF13B, have immune-related functions. Inflammatory responses were implicated in the patient with the WDFY4 mutation. Malfunction of the immune system and mitochondrial anomalies were shown in the patient with the TNFSF13B mutation. Prevalence of heterozygous SLC52A3 BVVL causative mutations and notable variability in expressivity of homozygous and heterozygous genotypes are being reported for the first time. Identification of WDFY4 and TNFSF13B as candidate causative genes supports conjectures on involvement of the immune system in BVVL and amyotrophic lateral sclerosis

Description of combined ARHSP/JALS phenotype in some patients with SPG11 mutations

Background: SPG11 mutations can cause autosomal recessive hereditary spastic paraplegia (ARHSP) and juvenile amyotrophic lateral sclerosis (JALS). Because these diseases share some clinical presentations and both can be caused by SPG11 mutations, it was considered that definitive diagnosis may not be straight forward. Methods: The DNAs of referred ARHSP and JALS patients were exome sequenced. Clinical data of patients with SPG11 mutations were gathered by interviews and neurological examinations including electrodiagnosis (EDX) and magnetic resonance imaging (MRI). Results: Eight probands with SPG11 mutations were identified. Two mutations are novel. Among seven Iranian probands, six carried the p.Glu1026Argfs*4-causing mutation. All eight patients had features known to be present in both ARHSP and JALS. Additionally and surprisingly, presence of both thin corpus callosum (TCC) on MRI and motor neuronopathy were also observed in seven patients. These presentations are, respectively, key suggestive features of ARHSP and JALS. Conclusion: We suggest that rather than ARHSP or JALS, combined ARHSP/JALS is the appropriate description of seven patients studied. Criteria for ARHSP, JALS, and combined ARHSP/JALS designations among patients with SPG11 mutations are suggested. The importance of performing both EDX and MRI is emphasized. Initial screening for p.Glu1026Argfs*4 may facilitate SPG11 screenings in Iranian patients. © 2020 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals LL

Evaluating the antioxidant and acetylcholinesterase inhibitory activities of some plants from Kohgiluyeh va Boyerahmad province, Iran

Background and objectives:Alzheimer’s disease (AD) is a neurodegenerative disorder. Nowadays, many investigations are performed to find new drugs for AD and medicinal plants are considered as one of the most important sources for developing new drugs. According to the role of oxidant agents and acetylcholinesterase enzyme (AChE) in AD, plants with antioxidant and AChE inhibition properties could be good candidates for AD studies. In the present investigation, acetylcholinesterase inhibition (AChEI) and antioxidant effects of some plants from Kohgiluye va Boyerahmad province of Iran have been determined. Methods: The plants collected from Kohgiluyeh va Boyerahmad province (56 species) were extracted with methanol by using maceration method. AChEI activity of the extracts was determined using Ellman method in 96-well microplates. Antioxidant activity was determined using DPPH and FRAP methods. Results: The results showed that aerial parts of Amygdalus scoparia had the highest AChEI effect (50% inhibition in concentration of 300 µg/mL). The plant also demonstrated suitable antioxidant effects. Epilobium minutiflorum found to be the most potent species for DPPH inhibition and reduction of ferric-TPTZ complex (IC50 3.6 µg/mL and FRAP value 335.0 mmol FeSO4.7H2O/100g Extract). Conclusion: Our results confirmed that almost all species with AChEI activity showed to be effective as potent antioxidant agents

Mesenchymal stem cells therapy protects fetuses from resorption and induces Th2 type cytokines profile in abortion prone mouse model

The imbalance of Th1/Th2 cytokines is well known in recurrent spontaneous abortion (RSA) mouse model. Mesenchymal stem cells (MSCs) possess potent immunoregulatory properties that could modulate the Th1 cytokine responses in benefit of Th2 types. In this study, we aimed to analyze the local and systemic balance of Th1/Th2 cytokines following MSCs therapy. Syngeneic adipose derived MSCs were administered to abortion prone mice during the implantation window. The abortion rate was determined and IL-4, IL-6, IL-12, IL-2, IFN-γ and GM-CSF gene expression was evaluated by Real-Time-PCR in decidual and placental tissues of pregnant mice at day 13.5 of pregnancy. Splenocytes of pregnant mice were co-cultured with mitomycin C treated paternal splenocytes and IL-2, IL-4, IL-10 and IFN-γ cytokines were measured in co-cultures supernatants by ELISA method. Proliferation response of female splenocytes to paternal antigens was also evaluated using the CFSE method. Our results showed a significant reduction in abortion rate following MSCs administration in abortion prone mice. We also observed a significant down-regulation of IL-2 and IFN-γ as well as up-regulation of IL-4 and IL-10 production from pregnant mouse splenocytes following MSCs therapy along with a significant reduction of splenocytes proliferation against paternal antigens. Our findings revealed that MSCs therapy increased the IL-4, IL-6, IL-10 and GM-CSF and at the same time decreased the IL-12, IL-2 and IFN-γ gene expression at feto-maternal interface. Here, we showed that MSCs therapy could modulate the systemic as well as local Th1/Th2 cytokines production along with protection of fetus from resorption in abortion prone mice. The fine balance of Th1/Th2 cytokine response could be considered as one of the possible mechanisms for fetal protection following MSCs therapy. © 2018 Elsevier B.V

Study on the seroprevalence of toxoplasmosis in sheep and goats of Tabriz area using the ELISA method

Toxoplasmosis is one of the most important zoonotic diseases and causes abortion in sheep and goats. In order to detect the slaughterhouse infection rate of toxoplasmosis in sheep and goats of Tabriz area, 186 blood sera samples were prepared and examined for IgG antibodies against toxoplasmosis using the ELISA method. Serological results indicated that the infection rate of Toxoplasma gondii was 18.3% with 13.45% of the infection rate related to sheep and 4.85% related to goats. The results indicate significant difference in the infection rate between sheep and goats of the region which could be related to the higher susceptibility of sheep to toxoplasmosis in comparison with goats. Based on the results of the study the infection rate in females was higher than males with the infection rate in male sheep being 11.5% while that of the females was 25.8% indicating signification difference in infection rates between males and females. The infection rate in male and female goats was 9.1% and 10.8% respectively and no significant difference in infection rate was observed between male and female goats

Evaluating the antioxidant and acetylcholinesterase inhibitory activity of three Centaurea species

Factors such as oxidative stress and reduced acetylcholine level have been implicated in Alzheimer’s disease (AD) pathology and recently there has been a trend towards natural product research to find potential sources of antioxidants and acetylcholinesterase inhibitors in the plants kingdom. Centaurea is a genus with about 500 species world wild, many of them have shown to possess biologic activity; Centaurea albonites, C. aucheri and C. pseudoscabiosa are three species which little investigation has been carried out about their biological properties. In the present study, the antioxidant and acetylcholinesterase inhibitory activity of the above mentioned species have been evaluated. The ability of the total extract and methanol fraction of the plants to scavenge free radicals has been assessed through DPPH radical scavenging assay, and the acetylcholinesterase inhibitory property has been evaluated by Ellman method. The total extract of all species exhibited moderate antioxidant activity whereas the extracts of C. pseudoscabiosa showed the strongest antioxidant property; its total extract also demonstrated the highest acetylcholinesterase inhibitory activity among the evaluated samples (19.2% inhibition). The results suggest the species as potential sources of natural antioxidants which could be focused in future studies of Alzheimer’s disease

Analysis of endometrial myeloid and lymphoid dendritic cells during mouse estrous cycle

This study was performed to evaluate the frequency and localization of endometrial myeloid (CD11c+ CD11b+) and lymphoid (CD11c+ CD8α+) dendritic cells (DCs) at different stages of murine estrous cycle. To address the systemic effect of ovarian hormones fluctuations during estrous cycle, the same variables were studied in splenic DCs as well. Stages of the estrous cycle of Balb/c mice were determined by examination of vaginal smears. Frozen sections of uterus and spleen at each stage of estrous cycle were stained for CD11c and MHC-II. Two-color immunohistochemistry was also carried out using anti-CD11c with one of the antibodies against CD11b, CD8α, CD86, and DEC-205. The average density of DCs and relative percentage of myeloid and lymphoid DCs (MDCs and LDCs) were determined at each stage of estrous cycle by morphometric analysis. Our results showed that DCs were present throughout the estrous cycle in mice endometrium, but their frequency was highest at estrus and lowest at proestrus (P < 0.005). The lymphoid subset of DCs was more prominent at estrus relative to those at other stages (P < 0.005). Conversely, the relative percentage of myeloid DCs at estrus was significantly lower compared to other stages (P < 0.005). Nearly all endometrial and splenic DCs expressed CD86 and MHC-II. At proestrus, and particularly at estrus, DCs were more concentrated subadjacent to the luminal and glandular epithelial layers with some scattered throughout the stroma whereas, at metestrus and diestrus, DCs were randomly distributed in stroma and around the glandular and luminal epithelial layers. The number and immunophenotype of splenic DCs were not statistically different between stages of estrous cycle. Our results suggest that endometrial but not splenic myeloid and lymphoid DCs are influenced by steroid hormones during estrous cycle. © 2006 Elsevier Ireland Ltd. All rights reserved

Cytotoxic activity of some ethnic medicinal plants from southwest of Iran

Background and objectives: Many people in ethnic groups of the world have trusted in plants for disease management and cure. Medicinal plants have always played a great role in the lives of Iranian people in the past and present and with no doubt in the future. Healers in different regions of Iran have been using medicinal herbs and one rich source for these cures is located in south-west of Iran, Kohgiluyeh va Boyer Ahmad province. Some species from this province have been selected for the present study and their cytotoxic activity has been evaluated. Methods: The methanol extracts of the 26 species were obtained by maceration and the extracts were investigated for cytotoxic activity in MTT assay. Results: The results revealed that four out of twenty six plants were toxic to MCF-7, A-549, HepG-2 and HT-29 cells. Conclusion: The findings of the present study specially the species with lower IC50 values (Eryngium billardieri and Nerium indicum) are suggested for further investigations in cancer studies

Deep geno- and phenotyping in two consanguineous families with CMT2 reveals HADHA as an unusual disease-causing gene and an intronic variant in GDAP1 as an unusual mutation

BACKGROUND: Charcot–Marie–Tooth (CMT) disease is a prevalent and heterogeneous peripheral neuropathy. Most patients affected with the axonal form of CMT (CMT2) do not harbor mutations in the approximately 90 known CMT-associated genes. We aimed to identify causative genes in two CMT2 pedigrees. METHODS: Neurologic examination, laboratory tests and brain MRIs were performed. Genetic analysis included exome sequencing of four patients from the two pedigrees. The predicted effect of a deep intronic mutation on splicing was tested by regular and real-time PCR and sequencing. RESULTS: Clinical data were consistent with CMT2 diagnosis. Inheritance patterns were autosomal recessive. Exome data of CMT2-101 did not include mutations in known CMT-associated genes. Sequence data, segregation analysis, bioinformatics analysis, evolutionary conservation, and information in the literature strongly implicated HADHA as the causative gene. An intronic variation positioned 23 nucleotides away from following intron/exon border in GDAP1 was ultimately identified as cause of CMT in CMT2-102. It was shown to affect splicing. CONCLUSION: The finding of a HADHA mutation as a cause of CMT is of interest because its encoded protein is a subunit of the mitochondrial trifunctional protein (MTP) complex, a mitochondrial enzyme involved in long chain fatty acid oxidation. Long chain fatty acid oxidation is an important source of energy for skeletal muscles. The mutation found in CMT2-102 is only the second intronic mutation reported in GDAP1. The mutation in the CMT2-102 pedigree was outside the canonical splice site sequences, emphasizing the importance of careful examination of available intronic sequences in exome sequence data