Characterization of Fasciola spp. in Myanmar on the basis of spermatogenesis status and nuclear and mitochondrial DNA markers

Fasciola spp. in Myanmar were characterized on the basis of spermatogenesis status and DNA markers of nuclear internal transcribed spacer 1 (ITS1) and mitochondrial NADH dehydrogenase subunit 1 (nad1). We collected 88 adult flukes from Yangon, Lashio, and Myitkyina. Spermatogenesis status was analyzed by the presence of sperm in the seminal vesicles, and 8 aspermic and 80 spermic flukes were detected. The flukes were identified on the basis of spermatogenesis status and ITS1 types which were analyzed by a PCR-RFLP method, and 80 spermic flukes were identified as F. gigantica. A very low detection rate of aspermic Fasciola sp. indicated that they are not established in Myanmar. In phylogenetic analyses, the 7 aspermic Fasciola sp. from Myitkyina displayed a haplotype in nad1 sequence, which was identical to that of aspermic Fasciola sp. from other Asian countries including China. Therefore, they were probably introduced from China through an infected domestic ruminant. On the other hand, 17 nad1 haplotypes detected in F. gigantica belonged to 2 clades unique to Myanmar, each with a distinct founder haplotype in a network analysis. This indicated a unique history of F. gigantica introduction into Myanmar involving ancient artificial movements of domestic ruminants.     

Time-Dependent Variations in Serum Nitrite, 6-Keto-Prostaglandin F1α and Thromboxane B2 Levels Induced by Lipopolysaccharide in Mice

Clinical features of certain immuno-inflammatory disorders exhibit time-dependent fluctuations, which could be related to circadian rhythmicity of proinflammatory mediator production. Many biologically active substances including nitric oxide (NO) and eicosanoids are released into the circulation in sepsis. Increased NO and eicosanoid levels have been reported to be responsible from death in septic shock. The aim of this study was to investigate the variations in the NO and eicosanoid production and mortality induced by bacterial endotoxin, lipopolysaccharide (LPS) injected either in the morning or in the evening. Experiments were performed on mice synchronised to 12 h light and 12 h dark (lights on at 09:00 h). Animals were injected intraperitoneally with LPS (10 mg/kg) at 09:00 (morning) and 21:00 h (evening) alone or in combination with aminoguanidine (NO synthase (NOS) inhibitor) (100 mg/kg) or indomethacin (cyclooxygenase (COX) inhibitor) (100 mg/kg). The serum was separated from blood samples obtained at nine different time points. Nitrite (stable product of NO), 6-keto-prostaglandin F_1α (6-keto-PGF_1α, stable product of prostacyclin) and thromboxane B₂ (TxB₂, stable product of thromboxane) concentrations in serum samples were measured. Serum nitrite levels showed a 24 h circadian rhythmicity depending on LPS injection time. Morning injection caused a peak after 15 h, while evening injection had two peaks after 9 and 18 h. The peak values obtained from morning and evening injections were significantly decreased by aminoguanidine and indomethacin. When LPS injected to mice in the morning and in the evening, it gradually increased the mortality rate within 24 h which could be abolished by aminoguanidine, but not indomethacin. Indomethacin-induced inhibition on LPS-induced nitrite levels was higher in the morning than in the evening. 6-keto-PGF_1α and TxB₂ levels were decreased by indomethacin when injected with LPS at both injection times, but not aminoguanidine. These results showed that there is an interaction between NO and eicosanoids, and LPS may produce different effects on NOS activity, but not eicosanoid production and mortality, depending on injection time in the experimental septic shock model in mice. Chronopharmacological manipulations of NOS and COX pathways and interactions between them could lead to novel therapeutic approaches for the treatment of septic shock.     

Activation of AMPK-Regulated CRH Neurons in the PVH is Sufficient and Necessary to Induce Dietary Preference for Carbohydrate over Fat

Download video (2MB)Help with mp4 files     

Intranasal ovalbumin immunotherapy with mycobacterial adjuvant promotes regulatory T cell accumulation in lung tissues

Allergen‐specific immunotherapy to induce T regulatory cells in the periphery has been used to treat allergic diseases. Mycobacteria can be used as an adjuvant for inducing T regulatory cells. However, it is unclear whether intranasal immunotherapy in combination with Mycobacteria adjuvant induces regulatory T cell differentiation and attenuates allergic responses in vivo. To investigate the role of intranasal ovalbumin (OVA) treatment alone and in combination with Mycobacteria vaccae, proportions of FoxP3⁺ regulatory T cells and anti‐inflammatory responses were evaluated in a murine model of asthma that was established in three groups of bicistronic Foxp3^EGFP reporter BALB/c mice. Before establishment of the asthma model, two groups of mice received intranasal OVA immunotherapy and one also received simultaneous s.c. M. vaccae. Expression of CD4⁺CD25⁺Foxp3^+EGFP+ T cells in the lung and spleen was analyzed by flow cytometry and the cytokine profiles of allergen‐stimulated lung and spleen lymphocytes assessed. The intranasal OVA immunotherapy group showed greater expression of CD4⁺CD25⁺Foxp3^+EGFP+ T cells in the spleen whereas in the group that also received M. vaccae such greater expression was demonstrated in the lung. Additionally, the proportion of IL‐10 and IFN‐γ‐secreting splenocytes was greater in the intranasal OVA + M. vaccae group. CD25 neutralization decreased CD4⁺Foxp3⁺ cells more than other groups. In parallel with this finding, production of IL‐10 and IFN‐γ was down‐regulated. Mucosal administration of OVA antigen results in a greater proportion of CD4⁺Foxp3⁺ T cells in the spleen. IL‐10 and IFN‐γ induced by intranasal OVA immunotherapy and M. vaccae administration is down‐regulated after CD25 neutralization.

     

Retinal Electrophysiological Effects of Intravitreal Bone Marrow Derived Mesenchymal Stem Cells in Streptozotocin Induced Diabetic Rats

Diabetic retinopathy is the most common cause of legal blindness in developed countries at middle age adults. In this study diabetes was induced by streptozotocin (STZ) in male Wistar albino rats. After 3 months of diabetes, rights eye were injected intravitreally with green fluorescein protein (GFP) labelled bone marrow derived stem cells (BMSC) and left eyes with balanced salt solution (Sham). Animals were grouped as Baseline (n = 51), Diabetic (n = 45), Diabetic+BMSC (n = 45 eyes), Diabetic+Sham (n = 45 eyes), Healthy+BMSC (n = 6 eyes), Healthy+Sham (n = 6 eyes). Immunohistology analysis showed an increased retinal gliosis in the Diabetic group, compared to Baseline group, which was assessed with GFAP and vimentin expression. In the immunofluorescence analysis BMSC were observed to integrate mostly into the inner retina and expressing GFP. Diabetic group had prominently lower oscillatory potential wave amplitudes than the Baseline group. Three weeks after intravitreal injection Diabetic+BMSC group had significantly better amplitudes than the Diabetic+Sham group. Taken together intravitreal BMSC were thought to improve visual function.     

Validation of G6PD Point-of-Care Tests among Healthy Volunteers in Yangon,Myanmar

Primaquine and other 8-amnoquinoline based anti-malarials can cause haemolysis in subjects with glucose-6-phosphate dehydrogenase (G6PD) deficiency. Correct diagnosis of G6PD status in patients is crucial for safe treatment of both relapsing stages of Plasmodium vivax and transmitting forms of Plasmodium falciparum. Lack of suitable point-of-care tests has hampered a much needed wide use of primaquine for malaria elimination. In this study we have assessed the performances of two qualitative tests, the fluorescent spot test (FST) and the G6PD CareStart test (CST), against the gold standard quantitative spectrophotometric assay in a population of 1000 random adult healthy volunteers living in Yangon, Myanmar. The prevalence of G6PD deficiency in the Bamar, Karen and in the whole sample set was 6.6% (10.1% in males), 9.2% (21.0% in males) and 6.8% (11.1% in males) respectively. The FST and CST showed comparable performances with sensitivity over 95% and specificity over 90%, however for cases with severe G6PD activity the FTS had improved performance. If used with a conservative interpretation of the signal, the CareStart test has the potential to be used in the field and, by allowing a wider use of primaquine, to help malaria elimination.     

Cytogenetic study of recurrent miscarriages and their parents

There is substantial evidence that genetic alterations are contributing factors to the risk for recurrent miscarriages. This study was conducted to determine the frequency and contribution of chromosomal abnormalities in miscarriages and in couples with recurrent miscarriages. We studied a total of 41 miscarriages and their parents with a history of 2–11 recurrent miscarriages. Chromosomal analysis from chorionic villus sampling (CVS) and fetal tissues were performed according to standard cytogenetic methods using G-banding technique. Major chromosomal aberrations and polymorphic variants were found in 51 and 4.8%, respectively. The chromosomal abnormalities were structural (34.4%) and numerical (65.1%) of which 26.1, 21.7, 8.7 and 8.7% were fetal sex aneuploid, triploid, mosaics and trisomic, respectively. Unbalanced and balanced rearrangements were found in 17.2 and 8.6% of all abnormalities, respectively. Major chromosomal abnormalities in couples were seen in 4.9%. The chromosomal abnormalities associated with pregnancy losses and recurrent miscarriages are mostly numerical ones. The incidence of balanced translocations found here is 4.9% which is near to the mode (about 3–6%) observed in the previous studies. Those frequencies are greater than in the general population (0.3%). This indicates that balanced translocations, seen in parents, have some importance in causing miscarriage. The major parental chromosomal aberrations are significantly associated with fetal wastage. Mosaicism should be taken into account for cytogenetic analyses of pregnancy losses. Thus, cytogenetic analyses should be recommended in couples with recurrent miscarriages, when clinical data fail to clarify the cause. The text was submitted by the authors in English.