Effects of Lucilia sericata on wound healing in streptozotocin-induced diabetic rats and analysis of its secretome at the proteome level.

The use of Lucilia sericata larvae on the healing of wounds in diabetics has been reported. However, the role of the excretion/secretion (ES) products of the larvae in treatment of diabetic wounds remains unknown. This study investigated whether application of the ES products of L. sericata on the wound surface could improve the impaired wound healing in streptozotocin-induced diabetic rats. Additional analysis was performed to understand proteome content of L. sericata secretome to understand ES contribution at the molecular level. For this purpose, full-thickness skin wounds were created on the backs of diabetic and control rats. A study was conducted to assess the levels of the ES-induced collagen I/III expression and to assay nuclear factor κB (NF-κB) (p65) activity in wound biopsies and ES-treated wounds of diabetic rat skin in comparison to the controls. The expression levels of collagen I/III and NF-κB (p65) activity were determined at days 3, 7, and 14 after wounding using immunohistological analyses and enzyme-linked immunosorbent assay technique. The results indicated that treatment with the ES extract increased collagen I expressions of the wound control and diabetic tissue. But the increase in collagen I expression in the controls was higher than the one in the diabetics. NF-κB (p65) activity was also increased in diabetic wounds compared to the controls, whereas it was decreased in third and seventh days upon ES treatment. The results indicated that ES products of L. sericata may enhance the process of wound healing by influencing phases such as inflammation, NF-κB (p65) activity, collagen synthesis, and wound contraction. These findings may provide new insights into understanding of therapeutic potential of ES in wound healing in diabetics.

Association of serum sex steroid levels and bone mineral density with CYP17 and CYP19 gene polymorphisms in postmenopausal women in Turkey.

Many clinical conditions, including osteoporosis, are associated with serum levels of sex steroids. Enzymes that regulate rate-limiting steps of steroidogenic pathways, such as CYP17 and CYP19, are also regarded as significant factors that may cause the development of these conditions. We investigated the association of two common polymorphisms, in the promoter region (T→C substitution) of CYP17 and exon 3 (G→A) of CYP19, with bone mineral density (BMD) in the lumbar spine and femoral neck and serum androgen/estradiol, in a case-control study of 172 postmenopausal women aged 62.3 ± 9.6 years (mean ± SD). The CYP17 TC genotype was significantly overrepresented in patients compared to controls, and TC genotype neck T-score and lumbar T-score values were significantly higher in patients compared to controls. CYP17 TC and TT genotype testosterone and DHEA-SO(4) levels were lower in patients compared to controls. All three genotypes of CYP19 had almost the same distribution among patients. The CYP19 AG genotype, however, was most frequent among controls. CYP19 lumbar BMD levels were close to each other among the different genotypes; however, AA and AG genotypes were significantly lower in patients. Testosterone and DHEA-SO(4) levels in the CYP19 GG genotype were higher compared to those of the other genotypes in patients but not in controls. CYP19 GA individuals had lower E(2) levels and lower BMD in controls and patients. Femoral neck BMD and lumbar T-score were also diminished with GA transition. In conclusion, CYP17 and CYP19 gene polymorphisms were found to be associated with osteoporosis in postmenopausal women in Turkey.

Heat shock proteins and myocardial protection during cardiopulmonary bypass.

This prospective randomized study investigated the effects of two different cardioplegia techniques on myocardial heat shock protein 70 (HSP70) mRNA levels. Patients undergoing elective coronary artery bypass grafting with cardiopulmonary bypass (CPB) were divided into two equal groups. All patients received the same anaesthesia. Myocardial preservation was achieved by delivering intermittent antegrade isothermic blood cardioplegia in one group and antegrade plus continuous retrograde isothermic blood cardioplegia in the other. Biopsies for measurement of HSP70 mRNA levels were taken from the right atria before surgical manipulation of the heart, and later from the same place following CPB. HSP70 mRNA levels were evaluated using quantitative real-time reverse transcription-polymerase chain reaction. Crossing-point values for HSP70 and ß-actin were used to evaluate up-regulation. There was a significant increase in HSP70 mRNA levels in response to CPB in both groups, but no significant between-group difference in HSP70 up-regulation. Further investigation is required to evaluate the correlation between the level of HSP induction and the degree of myocardial protection in more heterogeneous groups of patients.

Pancreatic islet derived stem cells can express co-stimulatory molecules of antigen-presenting cells.

BACKGROUND: Antigen-presenting cells (APCs) are crucial intermediates in the generation of both innate and specific immune responses. It has long been understood that some APCs are resident in islets in situ as well as after isolation. Our aim was to investigate the presence of molecules involved in antigen presentation in rat pancreatic islet-derived stem cells (PI-SCs). METHODS: We used immunocytochemistry and reverse transcription polymerization chain reaction to study immunophenotypic characteristics; pluripotent-related gene expressions; transcripts coding for antigen-presenting surface proteins CD40, CD80, CD86; and major histocompatibility complex class II in addition to genes with known antiapoptotic functions including mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK2), tumor necrosis factor alpha-induced protein 3 (TNFAIP3) interacting protein 1 (TNIP1) and BCL3 of the PI-SCs. RESULTS: Rat PI-SCs were negative for CD45 as demonstrated by flow cytometry and for CD31, CD34, and CD71 as demonstrated by immunocytochemistry. Therefore, there was no evidence of hematopoietic precursors in the cultures. OCT4, SOX2, and REX1 were expressed by rat PI-SCs. We determined the expression of genes for antigen-presenting surface proteins CD40 and CD80, and genes with known antiapoptotic functions including MAPKAPK2, TNIP1 and BCL3, besides the surface protein, CD80, by flow cytometry. CONCLUSION: Expression of these genes by rat PI-SCs implied that they could be involved in the regulation of immunity in islets, highlighting the influence of protective role-playing antiapoptotic mechanisms on pancreatic islet cells. This study offers the potential to understand the molecular mechanisms of a devastating disease, type-1 diabetes mellitus.

Nitrogen-fixation genes and nitrogenase activity in Clostridium acetobutylicum and Clostridium beijerinckii.

Several solvent-producing clostridia, including Clostridium acetobutylicum and C. beijerinckii, were previously shown to be nitrogen-fixing organisms based on the incorporation of 15N2 into cellular material. The key nitrogen-fixation (nif) genes, including nifH, nifD, and nifK for nitrogenase component proteins as well as nifE, nifN, nifB and nifV for synthesis of the iron-molybdenum cofactor (FeMoco) of nitrogenase, have now been identified in C. acetobutylicum or C. beijerinckii or both. The organization of these genes is similar to the distinctive pattern that was first observed in Clostridium pasteurianum, with the nifN and nifB genes fused into the nifN-B gene and with the nifV gene split into the nifVomega and nifValpha genes. The corresponding nif genes of these three clostridial species are highly related to each other. However, in the two solvent-producing clostridia, the nifH and nifD genes are interspersed by two glnB-like genes, which are absent in the corresponding region in C. pasteurianum. However, the nifN-B and nifVomega genes of C. pasteurianum are interspersed by the putative modA and modB genes (for molybdate transport), which are absent in the corresponding region in C. acetobutylicum. C. acetobutylicum and C. beijerinckii grew well under nitrogen-fixing conditions, and the acetylene-reducing activity of nitrogenase was measured in the two species. Acetone, butanol, and isopropanol production occurred in nitrogen-fixing cultures, but the peak of nitrogen-fixing activity preceded the active solventogenic phase.

Insecticide resistance in Anopheles sacharovi Favre in southern Turkey.

We report the resistance to 12 insecticides of specimens of Anopheles sacharovi, both in laboratory cultures and those collected in the malarious areas of Adana, Adiyaman, Antalya, Aydin, and Mugla in southern Turkey. Mortality was higher 24 h after exposure than immediately after exposure but was unaffected by temperature (24 degrees C or 29 degrees C) or the position of the test kit (horizontal or vertical). In Adana, Adiyaman and Antalya, A. sacharovi was susceptible only to malathion and pirimiphos-methyl. In Aydin it was susceptible to both these insecticides as well as to dieldrin, lambda-cyhalothrin, and etofenprox; and in Mugla it was susceptible to dieldrin, fenitrothion, lambda-cyhalothrin, cyfluthrin and etofenprox, as well as to malathion and pirimiphos-methyl.

Sandflies (Diptera: Psychodidae) associated with epidemic cutaneous leishmaniasis in Sanliurfa, Turkey.

As part of a project to study the possible impact of environmental change on health in southeastern Turkey, we evaluated sandfly species diversity, abundance, and habitat associations in an urban area where cutaneous leishmaniasis was undergoing epidemic re-emergence. Houses and caves in and around the city of Sanliurfa, Turkey, were sampled using mechanical aspirators, sticky papers, and CDC light traps. Of 1,649 sandflies captured, including 6 Phlebotomus and 1 Sergentomyia species, nearly all were P. papatasi (Scopoli) (967) or P. sergenti Parrot (674). Sandflies were active during June-September (hot dry season), but not during January (cool rainy season). Resting phlebotomines were abundant inside houses. Houses sampled in 3 neighborhoods with a high cutaneous leishmaniasis incidence (9-65 cases per 1,000 population) had > 10 times more flies than at a comparison site where few cases (0.2 per 1,000) have been reported. Results indicated that P. sergenti or P. papatasi were the probable vectors of cutaneous leishmaniasis during this outbreak and that control of these sandflies may eliminate transmission.

Species-specific detection of Leishmania in sandflies using an enzyme-linked immunosorbent assay.

A direct enzyme-linked immunosorbent assay (ELISA) using species-specific monoclonal antibodies was developed for the detection and identification of Leishmania in sandflies. A titration of mock-infected Phlebotomus papatasi showed that fewer than 2000 L. major promastigotes could be detected. The percentage of infected P. papatasi collected in the field, as determined by dissection, was compared to that revealed by the ELISA. Both methods gave similar results, irrespective of whether the flies were caught by sticky papers or light-traps. The percentage of infected flies determined by either method was also similar in experimentally infected colony reared sandflies. The ELISA can be carried out using multiple species-specific antibodies, and is as accurate as identification of infected sandflies by microscopical examination. The technique should be useful for identifying sandfly species involved in transmitting different species of Leishmania, and for rapid assessment of leishmanial infection rates in endemic regions.

Bloodfeeding behavior of Anopheles sacharovi in Turkey.

The feeding habits of Anopheles sacharovi under natural conditions and in feeding rooms were investigated by use of the gel diffusion technique. Mosquitoes were collected from various villages of Cukurova and also from feeding rooms especially prepared for these experiments. Human, cow, sheep, chicken, horse, and donkey were used as hosts in these rooms. The results showed that An. sacharovi is a zoophilic species. The females preferred donkey when human, cow, sheep, chicken, and horse were equally available. Their preference changed to horse, cow, and sheep in the absence of donkey. The host preference index (HPI) was always smaller than 1 for humans in habitats offering a choice of hosts. The human blood index was high only in human dwellings. In other habitats numbers of mosquitoes feeding on animals were higher than on humana. Although the human blood index was low, An. sacharovi is the principal human malaria vector in Turkey partly because a significant proportion of those resting in human dwellings have fed upon the occupants, and partly because of the uneven distribution of human and animal hosts.

Residual efficacy of pirimiphos methyl (Actellic) on Anopheles sacharovi in Cukurova, Turkey.

Village-scale trials of 50% emulsifiable concentrate (EC) and 40% wettable powder (WP) formulations of pirimiphos methyl (Actellic) were carried out against Anopheles sacharovi in Cukurova, Turkey. Susceptibility tests with wild caught, gonoactive and composite aged An. sacharovi over a range of chemical concentrations resulted in 100% mortality after exposure for 60 min to a 0.5% active ingredient concentration. Surface treatments of Actellic 50% EC at 0.9 g/m2 caused a significant decrease in parous rate and a 96.9% reduction in resting density. Persistence on concrete, wood, zinc and plywood was still high at the time of the second spray round, more than 7 wk postspray and ranged from 73% (zinc) to 98% (plywood). More than 50% mortality was still recorded 8 wk postspray using 1.6 g/m2 WP on wood, plywood, zinc and thatch substrates.

Development of Plasmodium vivax in Anopheles superpictus under experimental conditions.

The susceptibility of Anopheles superpictus for Plasmodium vivax was described quantitatively under laboratory conditions. Of the 697 laboratory females studied in 16 groups, 513 (73.5%) females took a bloodmeal, of which 88.4% developed ookinetes on day 1, 56.1% oocysts between days 3 and 11, and 52.5% sporozoites in the salivary gland on days 15 to 63 post-infection. Sporogony was completed in 10-11 days post-infection. There was no difference in the longevity of uninfected and infected females. Infected females survived an average of 30 days (maximum 63 days). Sporozoites survived up to 50 days in the salivary glands without any observable changes in structure and motility. These data indicate that An. superpictus is an efficient laboratory vector of P. vivax and should not be ignored in future entomological field studies.

Seasonal variation in populations of Anopheles maculipennis, Anopheles claviger and Culex pipiens in Turkey.

Seasonal variation in larval populations of three species of mosquitoes, Anopheles maculipennis, An. claviger and Culex pipiens, common in the vicinity of Ankara, were studied. Populations of An. maculipennis and Cx. pipiens disappeared in larval habitats by December but An. claviger, overwintering as larvae remained until the middle of March. Animal footprints are densely populated and are preferred by Cx. pipiens and An. claviger over larger water bodies.