Development of a facile system for mass production of Brugia malayi in a small-space laboratory

Brugia malayi is one of the important lymphatic filarial nematodes that cause elephantiasis and disability in humans in the Asian region. Mass production at any stage of this nematode in both small laboratory animal hosts and mosquito vectors is still necessary in order to continue various research aspects. This study elucidated on the use of nonblood feeding or the autogenous Ochlerotatus togoi (Thailand strain) and male Mongolian jird (Meriones unguiculatus) system. This has brought about a low-cost and highly-effective procedure for the mass production of blood containing microfilariae, infective (L₃) larvae, and adults of B. malayi under nonanimal-blood-feeding insectary and small-space animal-house conditions. The highly-infective rates (human-heparinized blood, 86.67–93.33; swine-heparinized blood, 83.33–96.67; bovine-heparinized blood, 76.67–80; chicken-heparinized blood, 73.33–76.67) and parasite loads (human-heparinized blood, 10.58–12.36; swine-heparinized blood, 8.40–10.38; bovine-heparinized blood, 9.75–9.91; chicken-heparinized blood, 3.41–4.65) of autogenous O. togoi to B. malayi and high numbers of adults recovered from ten B. malayi-infected male jirds (total?=?327, 16–52) are good supportive evidence. In addition, all special techniques required for succeeding in the establishment of a facile system regarding these matters are detailed.     

Molecular and cytogenetic evidence of three sibling species of the Anopheles barbirostris Form A (Diptera:Culicidae) in Thailand

Nine isoline colonies of Anopheles barbirostris Form A, derived from individual isofemale lines from Chiang Mai, Phetchaburi, and Kanchanaburi, were established in our insectary at Chiang Mai University. All isolines shared the same mitotic karyotype (X₁, X₂, Y₁). Molecular analysis of deoxyribonucleic acid (DNA) sequences and polymerase chain reaction (PCR) products of ITS2, COI, and COII regions revealed three distinct groups: A1 (Chiang Mai), A2 (Phetchaburi), and A3 (Kanchanaburi). Crossing experiments among the three groups exhibited strong reproductive isolation, producing low and/or non-hatched eggs, and inviable and/or abnormal development of the reproductive system of F₁-progenies. Asynaptic regions along the five polytene chromosome arms of F₁-hybrid larvae clearly supported the existence of three sibling species within A. barbirostris Form A, provisionally named species A1, A2, and A3.     

Salivary gland proteome of the human malaria vector, Anopheles campestris-like (Diptera: Culicidae)

Anopheles campestris-like is proven to be a high-potential vector of Plasmodium vivax in Thailand. In this study, A. campestris-like salivary gland proteins were determined and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis, and nano-liquid chromatography-mass spectrometry. The total amount of salivary gland proteins in the mosquitoes aged 3–5 days was approximately 0.1?±?0.05 μg/male and 1.38?±?0.01 μg/female. SDS-PAGE analysis revealed at least 12 major proteins found in the female salivary glands and each morphological region of the female glands contained different major proteins. Two-dimensional gel electrophoresis showed approximately 20 major and several minor protein spots displaying relative molecular masses from 10 to 72 kDa with electric points ranging from 3.9 to 10. At least 15 glycoproteins were detected in the female glands. Similar electrophoretic protein profiles were detected comparing the male and proximal-lateral lobes of the female glands, suggesting that these lobes are responsible for sugar feeding. Blood-feeding proteins, i.e., putative 5′-nucleotidase/apyrase, anti-platelet protein, long-form D7 salivary protein, D7-related 1 protein, and gSG6, were detected in the distal-lateral lobes (DL) and/or medial lobes (ML) of the female glands. The major spots related to housekeeping proteins from other arthropod species including Culex quinquefasciatus serine/threonine-protein kinase rio3 expressed in both male and female glands, Ixodes scapularis putative sil1 expressed in DL and ML, and I. scapularis putative cyclophilin A expressed in DL. These results provide information for further study on the salivary gland proteins of A. campestris-like that are involved in hematophagy and disease transmission.     

Efficacy and safety of conventional antiviral agents in preventive strategies for cytomegalovirus infection after kidney transplantation: a systematic review and network meta-analysis

Cytomegalovirus (CMV) infection is common in kidney transplantation (KT). Antiviral-agents are used as universal prophylaxis. Our purpose aimed to compare and rank efficacy and safety. MEDLINE, Embase, SCOPUS, and CENTRAL were used from inception to September 2020 regardless language restriction. We included randomized clinical trials (RCTs) comparing the CMV infection/disease prophylaxis among antiviral-agents in adult KT recipients. Of 24 eligible RCTs, prophylactic valganciclovir (VGC) could significantly lower the overall CMV infection and disease risks than placebo with pooled risk differences (RDs) [95% confidence interval (CI)] of −0.36 (−0.54, −0.18) and −0.28 (−0.48, −0.08), respectively. Valacyclovir (VAC) and ganciclovir (GC) significantly decreased risks with the corresponding RDs of −0.25 (−0.32, −0.19) and −0.30 (−0.37, −0.22) for CMV infection and −0.26 (−0.40, −0.12) and −0.22 (−0.31, −0.12) for CMV disease. For subgroup analysis by seropositive-donor and seronegative-recipient (D+/R−), VGC and GC significantly lowered the risk of CMV infection/disease with RDs of −0.42 (−0.84, −0.01) and −0.35 (−0.60, −0.12). For pre-emptive strategies, GC lowered the incidence of CMV disease significantly with pooled RDs of −0.33 (−0.47, −0.19). VGC may be the best in prophylaxis of CMV infection/disease follow by GC. VAC might be an alternative where VGC and GC are not available.     

Analysis of female salivary gland proteins of the Anopheles barbirostris complex (Diptera: Culicidae) in Thailand

Electrophoretic protein profiles of female salivary glands of five sibling species within the Anopheles barbirostris complex, namely A. barbirostris species A1 (Forms A, B, and D), A2, A3, and A4 and Anopheles campestris-like (Forms B and E), were analyzed. At least eight major and several minor protein bands were detected in the glands of each species, of which each morphological region contained different major proteins. The protein profiles distinguished the five sibling species. The variability in major proteins among species was observed in the 40–48, 32–37, and 10–18 kDa ranges. No difference in protein profiles was found in different cytogenetic forms. Polymorphism of the protein profiles within species was only noted in species A4. The lowest major protein (marker) band of each species showed remarkably different relative mobility on SDS–polyacrylamide gels. NanoLC-MS analysis revealed that the marker protein of some species matched with a protein involving in blood feeding, gSG6, of Anopheles gambiae and Anopheles freeborni. These results might be useful for construction of an additional tool to distinguish the five sibling species and lead to further study on the evolution of blood feeding and pathogen transmission.     

Cytogenetic and molecular evidence for an additional new species within the taxon Anopheles barbirostris (Diptera: Culicidae) in Thailand

ITS2 DNA sequences of 42 isoline colonies of Anopheles barbirostris species A1 and A2 were analyzed and a new genetic species, temporarily designated as species A4 (Chiang Mai), was revealed. The large sequence divergences of the ITS2 (0.116-0.615), COI (0.023–0.048), and COII (0.030–0.040) genes between A. barbirostris species A4/A1 (Chiang Mai), A4/A2 (Phetchaburi), A4/A3 (Kanchanaburi), and A4/Anopheles campestris-like Form E (Chiang Mai) provided good supporting evidence. Species A1, A2, A3, and A4 share a mitotic karyotype of Form A (X₁, X₂, Y₁). Crossing experiments between species A4 and the other four species yielded strong reproductive isolation producing few and/or non-hatched eggs and inviable and/or abnormal development of the reproductive system of F₁ progenies. Moreover, available F₁ hybrid larvae showed asynaptic polytene chromosome arms. Hence, molecular and cytogenetic evidence strongly support the existence of A. barbirostris species A4, which is more closely related to A. campestris-like Form E than to species A1, A2, and A3. Additionally, crossing experiments among 12 and seven isolines of different cytological forms of species A1 (A, B, C, D) and A2 (A, B), respectively, yielded fertile and viable F₁ progenies. Thus, different karyotypic forms occurring in natural populations of species A1 and A2 merely represent intraspecies variation of sex chromosomes due to the extra blocks of heterochromatin.     

A review of the etiologies,clinical characteristics,and treatment of canities

Hair pigmentation is regulated by follicular melanogenesis, in which the process consists of melanin formation and transfer to keratinocytes in the hair shaft. Human hair follicles contain two types of melanin: the brown-black eumelanin and yellow-red pheomelanin. Eumelanin is commonly present in black and brown hair while pheomelanin is found in auburn and blonde hair. Hair follicle melanogenesis is under cyclical control and is concurrently coupled to hair growth. Many factors including intrinsic and extrinsic factors affect the follicular melanogenesis. Though many studies have been conducted to identify the pathogenesis and regulation of hair pigmentation, the etiology of canities and hair pigmentation is still unclear. The pathogenesis of canities or gray hair is believed to occur either from insufficient melanin formation due to melanocyte degeneration or a defect in melanosomal transfer. Canities is an aging sign which often interferes with one's socio-cultural adjustment. On the other hand, premature canities correlate with diseases such as osteopenia and cardiovascular disease. Risk factors associated with canities are not only genetic but also external causes. For example, smoking, alcohol consumption, and stress are among the most common factors. Camouflage techniques are still used as the primary treatment of canities. Further treatments for canities are being developed to achieve the true reversal of hair pigmentation.     

Double-Filtration Plasmapheresis Plus Low-Dose Anti-thymocyte Globulin and Tacrolimus in Asian Living-Donor Kidney Transplantation With Donor-Specific Anti-HLA Antibody

BackgroundPretransplant desensitization protocols, including plasmapheresis, intravenous immunoglobulin, induction antibody therapy, and intensive maintenance immunosuppression, are generally employed in kidney transplant recipients who have positive status for donor-specific anti-HLA antibody (DSA). To avoid serious infectious complications, the authors designed a novel low-dose protocol in Thai patients undergoing DSA+ living-related kidney transplantation (LRKT).MethodsA retrospective cohort study of the patients who underwent DSA+ LRKT was conducted. The novel protocol consisted of 3 to 5 sessions of pretransplant double-filtration plasmapheresis (DFPP) with or without low-dose intravenous immunoglobulin together with low-dose anti-thymocyte globulin (ATG) induction (1-1.5 mg/kg/d for 3-4 days) and low-dose tacrolimus (Tac) (trough level 5-10 ng/mL), mycophenolate, and prednisolone.ResultsThe study included 17 patients. The lymphocyte crossmatch via complement-dependent cytotoxicity was negative in 12 patients and positive for B cell immunoglobulin M in 5 patients. The novel desensitization protocol resulted in a decrease of at least 50% of DSA mean fluorescence intensity from baseline (from 4320 ± 549 before DFPP to 1601 ± 350 before transplantation, P < .005) and successful kidney transplantation with good allograft function in all cases. Early DSA rebound was observed in 3 patients after transplantation, and kidney biopsy revealed subclinical antibody-mediated rejection in 1 patient and diffuse C4d staining without cell infiltration in 2 patients. There were good long-term outcomes in patient and graft survival (100% and 94.1%, respectively). Only 1 allograft loss occurred because of nonadherence. The majority of patients have stable allograft function with serum creatinine less than 1.5 mg/dL. However, infections, including CMV and other organisms, were commonly observed.ConclusionsDesensitization protocol with DFPP, low-dose ATG, and Tac provides excellent outcomes in living donor kidney transplantation in highly sensitized Asian populations.