Big flies, small repeats: the "Thr-Gly" region of the period gene in Diptera

The region of the clock gene period (per) that encodes a repetitive tract of threonine-glycine (Thr-Gly) pairs has been compared between Dipteran species both within and outside the Drosophilidae. All the non- Drosophilidae sequences in this region are short and present a remarkably stable picture compared to the Drosophilidae, in which the region is much larger and extremely variable, both in size and composition. The accelerated evolution in the repetitive region of the Drosophilidae appears to be mainly due to an expansion of two ancestral repeats, one encoding a Thr-Gly dipeptide and the other a pentapeptide rich in serine, glycine, and asparagine or threonine. In some drosophilids the expansion involves a duplication of the pentapeptide sequence, but in Drosophila pseudoobscura both the dipeptide and the pentapeptide repeats are present in larger numbers. In the nondrosophilids, however, the pentapeptide sequence is represented by one copy and the dipeptide by two copies. These observations fulfill some of the predictions of recent theoretical models that have simulated the evolution of repetitive sequences.      

Evolutionary origin of human and primate malarias: evidence from the circumsporozoite protein gene

We have analyzed the conserved regions of the gene coding for the circumsporozoite protein (CSP) in 12 species of Plasmodium, the malaria parasite. The closest evolutionary relative of P. falciparum, the agent of malignant human malaria, is P. reichenowi, a chimpanzee parasite. This is consistent with the hypothesis that P. falciparum is an ancient human parasite, associated with humans since the divergence of the hominids from their closest hominoid relatives. Three other human Plasmodium species are each genetically indistinguishable from species parasitic to nonhuman primates; that is, for the DNA sequences included in our analysis, the differences between species are not greater than the differences between strains of the human species. The human P. malariae is indistinguishable from P. brasilianum, and P. vivax is indistinguishable from P. simium; P. brasilianum and P. simium are parasitic to New World monkeys. The human P. vivax-like is indistinguishable from P. simiovale, a parasite of Old World macaques. We conjecture that P. malariae, P. vivax, and P. vivax-like are evolutionarily recent human parasites, the first two at least acquired only within the last several thousand years, and perhaps within the last few hundred years, after the expansion of human populations in South America following the European colonizations. We estimate the rate of evolution of the conserved regions of the CSP gene as 2.46 x 10(-9) per site per year. The divergence between the P. falciparum and P. reichenowi lineages is accordingly dated 8.9 Myr ago. The divergence between the three lineages leading to the human parasites is very ancient, about 100 Myr old between P. malariae and P. vivax (and P. vivax-like) and about 165 Myr old between P. falciparum and the other two.      

Excitation of System I and System II in Photosynthesis as a Possible Control Mechanism of Glycolate Metabolism in the Blue-Green Alga Anacystis nidulans

Photosynthetic enhancement studies performed at 619 nm (excitation of Systems I and II) and at 446 nm (mainly excitation of System I) revealed an 18% photosynthetic enhancement simultaneously with a 31% reduction in glycolate excretion. This observation supports the hypothesis that some glycolate may be consumed in an oxidation process associated with System I when System II is poorly excited and the supply of electrons from the water splitting process of photosynthesis is low.     

The anabolic action of intermittent parathyroid hormone on cortical bone depends partly on its ability to induce nitric oxide‐mediated vasorelaxation in BALB/c mice

There is strong evidence that vasodilatory nitric oxide (NO) donors have anabolic effects on bone in humans. Parathyroid hormone (PTH), the only osteoanabolic drug currently approved, is also a vasodilator. We investigated whether the NO synthase inhibitor L‐NAME might alter the effect of PTH on bone by blocking its vasodilatory effect. BALB/c mice received 28 daily injections of PTH[1–34] (80 µg/kg/day) or L‐NAME (30 mg/kg/day), alone or in combination. Hindlimb blood perfusion was measured by laser Doppler imaging. Bone architecture, turnover and mechanical properties in the femur were analysed respectively by micro‐CT, histomorphometry and three‐point bending. PTH increased hindlimb blood flow by >30% within 10 min of injection (P < 0.001). Co‐treatment with L‐NAME blocked the action of PTH on blood flow, whereas L‐NAME alone had no effect. PTH treatment increased femoral cortical bone volume and formation rate by 20% and 110%, respectively (P < 0.001). PTH had no effect on trabecular bone volume in the femoral metaphysis although trabecular thickness and number were increased and decreased by 25%, respectively. Co‐treatment with L‐NAME restricted the PTH‐stimulated increase in cortical bone formation but had no clear‐cut effects in trabecular bone. Co‐treatment with L‐NAME did not affect the mechanical strength in femurs induced by iPTH. These results suggest that NO‐mediated vasorelaxation plays partly a role in the anabolic action of PTH on cortical bone. © 2016 The Authors. Cell Biochemistry and Function published by John Wiley & Sons, Ltd.     

Repeated immunostaining of the same tissue section using alkaline phosphatase as a reporter

One can determine the best dilution of a primary antibody for immunohistochemistry that uses horseradish peroxidase conjugated to a secondary antibody by testing increasing concentrations sequentially on the same tissue section. When the same tissue section is incubated repeatedly with increasing concentrations of primary antibodies to epithelial membrane antigen, smooth muscle α-actin, or vimentin using alkaline phosphatase conjugated to a secondary antibody as the reporter, the best staining was obtained with a less concentrated primary antibody than was optimal for a single staining test. The best concentration of primary antibody for single run staining using an alkaline phosphatase reporting system is usually four times the best concentration for staining with multiple runs. The optimal concentration can be determined by denaturing the residual alkaline phosphatase and extracting residual stain by incubating the section in 4:1 diglyme:phosphate buffered saline for 20 min at 80^o C between tests of primary antibody concentrations. I tested the method for four chromogens from one supplier and one chromogen from a different supplier.     

A revision of chiggers in the vulgaris group (Trombiculidae: Neotrombicula)

A revision of chigger mites species, being closely related to Neotrombicula vulgaris (Schluger, 1955), is given. 2 new species are described: N. aideriensis sp. n. from Western Kopetdagh and N. macrovulgaris sp. n. from Western Caucasus. N. aideriensis sp. n. differs from all other species of the vulgaris group by presence of single barb on the lateral seta of palpal tibia in most specimens. This species differs also from N. vulgaris by lesser width of scutum, shorter setae of scutum and idiosoma and by longer legs. N. macrovulgaris sp. n. differs from all other species of the vulgaris group by longer legs, more numerous idiosomal setae and by larger scutum. N. vulgaris is recorded for the first time in Bulgaria, Turkmenistan (western Kopet-Dag) and Krasnodar Territory, N. baschkirica Kudryashova, 1998--in Chuvashia, Kirov Province, Komi Republic and Tyumen' Province, N. kharadovi Kudryashova, 1998--in Russia (Altai Territory) and in Karaganda Province (Kazakhstan). The latter species is reported from several new hosts. Variation of morphometric parameters in the vulgaris group are investigated by the methods of multivariate analysis, the pattern of correlations between them is shown. A complex of diagnostic features in the group is revised. Discriminant functions produced by the computer program DIADIS (A. L. Lobanov, ZI RAS) allows to determine confidently representatives of the vulgaris group. Ecogeographical component of intraspecific variability and character variance at the specific level is revealed in vulgaris group. N. macrovulgaris characterized by the largest scutum, numerous setae and long legs was found in the most rigorous, cold and damp climate (alpine zone of western Caucasus). On the other hand, N. kharadovi, which have the most fine scutum and shortest setae, inhabits rather dry and warm Middle Asia and neighbouring territories. The sample of N. vulgaris collected in the steppe part of Stavropol' Territory differed from the material collected in mountain forests of Western Caucasus by low values of the most part of measurements. N. kharadovi samples from plains of Middle Asia differed from samples of this species collected in western Pamirs, central Kazakhstan and Altay Territory by more fine scutum and shorter setae.     

Variability in Leptotrombidium europaeum and two new related chigger mite species (Acari: Trombiculidae) from Caucasus

Two new chigger mite species closely related to Leptotrombidium europaeum (Daniel et Brelih, 1959) are described from small mammals collected in Caucasus and Transcaucasia. L. alanicum sp. n. differs from L. europaeum in having shorter legs (TaIII = 61-81, Ip = 734-927 versus 72-90, and 855-1017), shorter scutal and idiosomal setae (D(min) = = 30-45, D(max) = 48-67, H = 59, PL = 58 versus 40-52, 54-69, 64, 63), slightly smaller scutum (AP = 25, SD = 47, PW = 89 versus 28, 50, 91), and more numerous idiosomal setae (87 versus 81). L. montanum sp. n. differs from L. europaeum in having more numerous idiosomal setae (102 versus 81), longer scutal and idiosomal setae (AM = 61, AL = 44, H = 68, D(max) = 66 versus 56, 41, 64, 62), thicker legs (TaW = 19 versus 18), and broader scutum (PW = 95 versus 91). Exact identification of both new species is possible only using classification functions constructed by means of discriminant analysis. These three Leptotrombidium species expose sympatric distribution in Daghestan (Eastern Caucasus). L. alanicum and L. montanum also occurred together in Krasnodar Territory (Western Caucasus). Each of these species includes a number of local geographical forms precisely distinguished from each other. Morphometric differences between L. alanicum and L. montanum agree with eco-geographical rules being previously found in chigger mites from other genera. However, differences between local forms of these species show other tendencies directed controversially in part. Therefore it is probable that interspecific differences in this case correspond to the variability which took place in the process of speciation.     

Intraspecific variability and sympatry in closely related chigger mite species of the genus Hirsutiella (Acari: Trombiculidae)

Intraspecific variability in three closely related chigger mite species of the genus Hirsutiella Schluger et Vysotzkaya, 1970, H. steineri (Kepka, 1966), H. llogorensis (Daniel, 1960), and H. alpina Stekolnikov, 2001 has been studied based on materials collected in Caucasus and Turkey. It is established that both H. steineri and H. llogorensis include Western Caucasian and Asian forms, the first one being larger than the second one. Western Caucasian samples of H. steineri are also split into large and small forms. The large form inhabits screes with larvae occurring mainly on snow voles, and the small form inhabits meadows and forests with larvae parasitizing snow voles as well as mice of the genus Apodemus and pine voles. Asian population of H. steineri include small low-mountain form which is hard to distinguish from H. llogorensis. In the light of the new data on variability, morphological border between these two species are specified. Correlations between some characters and altitude above sea level are shown within the Western Caucasian and Asian forms of both species, and also in H. alpina. All the three species can occur together on the same individual hosts, but they have different sets of main hosts and different distribution among biotopes in the area of sympatria. The border between H. steineri and H. llogorensis in the places of sympatria can be indistinct owing to the presence of small ecological forms of H. steineri. Our results give a basis for the construction of alternative hypotheses concerning the processes of speciation in trombiculids. Chiggers species could be formed in allopatric way, on the base of such geographical forms, as Western and Asian forms of the Caucasian Hirsutiella species, but they could also be formed in sympatric way, distributing among neighbouring biotopes (e.g., screes and meadows, or screes and forests), as large and small forms of Western Caucasian H. steineri do.     

土壤中棉花黄萎病菌SYBR Green Ⅰ荧光RT-PCR定量检测技术研究

为实现田间土壤棉花黄萎病菌的早期检测,建立了土壤中棉花黄萎病菌的SYBR Green I荧光定量PCR检测方法。以含342bp PCR扩增产物的阳性质粒为参考,构建了标准曲线,并对该曲线的特异性、敏感性、可重复性进行了评价。结果表明,该方法具有快速、特异性强、敏感度高等特点。检测范围在3.8×103-3.8×108copies/μL之间有良好的线性关系,相关系数R2为0.996,扩增效率为101.5%,灵敏度比常规PCR方法高102倍。