Methods to study maternal regulation of germ cell specification in zebrafish.

The process by which the germ line is specified in the zebrafish embryo is under the control of maternal gene products that were produced during oogenesis. Zebrafish are highly amenable to microscopic observation of the processes governing maternal germ cell specification because early embryos are transparent, and the germ line is specified rapidly (within 4-5h post fertilization). Advantages of zebrafish over other models used to study vertebrate germ cell formation include their genetic tractability, the large numbers of progeny, and the easily manipulable genome, all of which make zebrafish an ideal system for studying the genetic regulators and cellular basis of germ cell formation and maintenance. Classical molecular biology techniques, including expression analysis through in situ hybridization and forward genetic screens, have laid the foundation for our understanding of germ cell development in zebrafish. In this chapter, we discuss some of these classic techniques, as well as recent cutting-edge methodologies that have improved our ability to visualize the process of germ cell specification and differentiation, and the tracking of specific molecules involved in these processes. Additionally, we discuss traditional and novel technologies for manipulating the zebrafish genome to identify new components through loss-of-function studies of putative germ cell regulators. Together with the numerous aforementioned advantages of zebrafish as a genetic model for studying development, we believe these new techniques will continue to advance zebrafish to the forefront for investigation of the molecular regulators of germ cell specification and germ line biology.

[HIV-1 genetic variants in the Asian part of Russia: a study (2005-2010)].

The HIV-1 genetic variants circulated in the Asian part of the Russian Federation in 2005-2010 were studied. The samples of HIV-1 (427 in total) were collected in Khabarovsk, Magadan, Kurgan, Krasnoyarsk, Noyabr'sk, Yakutsk, Altay, and Tyva. Sequencing of some genome regions followed by the phylogenetic analysis or specific Internet resource sampling were used as the main methods of the HIV subtyping. The domination of the IDU-A HIV-1 genetic variant typical of HIV-infection epidemic in Russia was shown in all regions tested in 2005-2010. This variant prevailed both in IDUs and heterosexuals. In addition to IDU-A, some other HIV-1 genetic variants were found among them: subtype B and recombinant CRF03_AB. The HIV-1 genetic polymorphism in Russia was found to be low. An increase in the genetic distance among studied de novo samples was noted in the Asian part of Russia in 2005-2010 (26-68%) as compared to the European variants in 1996-1999 (10%).

Inactivation of M-phase promoting factor at exit from first embryonic mitosis in the rat is independent of cyclin B1 degradation.

Exit from M-phase and completion of cell division requires inactivation of M-phase promoting factor (MPF), a heterodimer composed of the regulatory cyclin B1 and the catalytic p34cdc2 kinase. Inactivation of MPF is associated with cyclin B1 degradation that is brought about by the ubiquitin-proteasome pathway. Our study examined the role of the proteasome in the first mitosis of rat embryos and its participation in the regulation of cyclin B1 degradation and MPF inactivation. We show that in the early zygote the proteasome is evenly distributed in the ooplasm and the nucleus, whereas during mitosis it accumulates on the spindle apparatus. We further demonstrate that inhibition of proteasomal catalytic activity prevents 1-cell embryos from undergoing mitosis. This mitotic arrest is associated with the presence of relatively high amounts of cyclin B1, which unexpectedly does not result in elevated MPF activity. Our findings strongly imply that completion of the first embryonic division depends on proteasomal degradation and that cyclin B1 is included among its target proteins. They also provide the first evidence that MPF inactivation at this stage of development is not solely dependent upon cyclin B1 degradation and is insufficient to allow the formation of the 2-cell embryo.

Single-neuron responses to emotional visual stimuli recorded in human ventral prefrontal cortex.

Both lesion and functional imaging studies in humans, as well as neurophysiological studies in nonhuman primates, demonstrate the importance of the prefrontal cortex in representing the emotional value of sensory stimuli. Here we investigated single-neuron responses to emotional stimuli in an awake person with normal intellect. Recording from neurons within healthy tissue in ventral sites of the right prefrontal cortex, we found short-latency (120-160 ms) responses selective for aversive visual stimuli.

Granisetron and ondansetron for chemotherapy-related nausea and vomiting.

The efficacy of granisetron and ondansetron, two serotonin antagonists, as antiemetic agents was studied and compared in the first 24 h follow, by chemotherapy administration. Sixty cancer patients took part in this randomized prospective study. The patients were categorized according to the chemotherapy which was administered. Patients in group A received combinations including the highly emetogenic agents, cisplatinum or adriamycin, patients in group B received regimens which included neither of these agents. Both serotonin antagonists were highly effective, only 9 (16%) out of 58 patients suffered from vomiting, 7 (25%) of the patients on ondansetron and 2 (7%) on granisetron. Splitting the patients into both groups supported the above results. In group A (adriamycin-cisplatinum) only 7 patients (22%) reported vomiting, 5 (33%) on ondansetron vs 2 (12%) on granisetron. In group B, only 2 patients (8%) suffered from vomiting, both patients (15%) on ondansetron while no vomiting was seen in the granisetron patients. Similar results were obtained in preventing nausea in these patients. We conclude that both serotonin antagonists are effective in preventing nausea and vomiting in patients receiving chemotherapy. In all groups tested this study granisetron appeared to be superior, although, this difference failed to reach statistical significance due to the small number of patients analyzed. However, there was no difference in cost effectiveness between the regimens.

Q fever endocarditis in Israel and a worldwide review.

The worldwide epidemiology and population-based incidence of Q fever endocarditis (QFE) have been less well studied than those for uncomplicated Q fever. An exhaustive literature review revealed 408 patients with QFE reported between 1949 and 1994, mostly from 3 large geographic areas. Underlying valvular heart disease was almost invariably present, and 38% had prosthetic valves. The most common clinical manifestations were fever and congestive heart failure. The mortality rate dropped over the years from 65% to 25%, but a meta-analysis of published data showed the death rate to be significantly lower among patients receiving combination therapy (12/65, 18%), as compared to patients treated with tetracycline alone (18/41, 44%, p = 0.005). A 10-year (1983-1992) retrospective nationwide survey of QFE in Israel revealed 35 patients with QFE, representing an annual incidence of 0.75 cases per 1 million population. Underlying heart disease, clinical manifestations and outcome in the Israeli group were not substantially different from those described in the world literature. The current state-of-the-art clinical approach includes early diagnosis, prompt initiation of combination therapy for at least 3 years, and long-term clinical and serologic follow-up. Adherence to these rules might have contributed to the improved prognosis in recent years.

[Patho- and morphogenesis of pyo-inflammatory diseases of the adnexa uteri]. / Pato- i morfogenez gnoinykh vospalitel'nykh zabolevanii pridatkov matki.

The uterine appendages in 87 females aged from 17 to 53 years were surgically removed because of inefficacy of conservative treatment for purulent inflammation. There were signs of chronic recurrent inflammation in the uterine tubes or in the tubes and ovaries as shown by the operative material studied light and electron microscopically, immunomorphologically. Immune mechanisms as well as structural changes (reduction, vasculitis) of the vascular bed and muscular layer (sclerosis) of the tubes played a role in the pathogenesis of chronic inflammation. The detection of the antibodies against Chlamydobacterials in 60% of the patients implies their role in pathogenesis of chronic inflammation of the uterine appendages.

[The effect of recombinant interleukin-2 on the course of experimental staphylococcal peritonitis in mice]. / Vliianie rekombinantnogo interleikina-2 na techenie éksperimental'nogo stafilokokkovogo peritonita u myshei.

The effect of RIL-2 on the survival of mice with S. aureus--induced peritonitis was studied. Animals received bacterial suspension and RIL-2 as following: bacteria--on days 0, +2, RIL-2--day 0 (group 1); bacteria--days 0, +4, RIL-2--days 0, +2 (group 2); bacteria--days 0, +6, RIL-2--days 0, +2, +4 (group 3). RIL-2 exerted no protective effect in group 1. However, in groups 2 and 3, where the control animals survival was, resp., 56% and 38%, the RIL-2 treatment increased survival up to, resp., 84% and 70%. Antibiotics given instead of RIL-2 in analogous regimen decreased the survival in group 3 to the level of 25%. Thus, RIL-2 proved to be a potent therapeutic agent in the 2nd of 3d studied models of S. aureus--induced peritonitis in mice. The perspectives of RIL-2 use in the treatment of bacterial peritonitis, including porous ones, and of the immunodepression--aggravated conditions are discussed.

Recombinant interleukin-2 significantly increases the survival of mice with peritonitis, but not acute Staphylococcus aureus peritoneal infection.

The effect of recombinant interleukin 2 (rIL-2) on survival of mice with peritonitis and acute Staphylococcus aureus strain 5/2 infection was studied. rIL-2 was ineffective in the case of acute infection when administered simultaneously with LD95 dose of bacteria. The antibiotics (gentamycin or a combination of penicillin and streptomycin) administered in the same fashion cured 100% of animals. rIL-2 proved to be a potent healing agent in the two of three models of S aureus peritonitis. In this case animals received bacteria at days 0 and 2, 4, or 6. rIL-2 was injected at day 0 (group 1), days 0 and 2 (group 2), and days 0, 2, and 4 (group 3). Treatment with rIL-2 was ineffective in group 1; however, in groups 2 and 3 rIL-2 increased the survival up to 90% (in comparison with 30% in the untreated animals of group 2 and 64% in group 3). On the contrary, administration of antibiotics instead of rIL-2 in the group 3 decreased survival to 25%. The perspectives of rIL-2 use in the treatment of bacterial peritonitis, including purous ones, and the cases complicated by immunodepression, are discussed.

[The controversial aspects of granulomatous inflammation]. / Spornye aspekty granulematoznogo vospaleniia.

The necessity of a new concept of nontumorous benign monocytosis is brought forward for discussion. It is suggested to include into the group of "granulomatous inflammation" chronic inflammation with epithelioid cell granulomas. Species peculiarities of rodents (mice and rats) are underlined, i.e. their capacity to form liver macrophagal proliferates as a response to various stimuli including antigenic and acute infectious ones.

[Influence of recombinant interleukin-2 on the course and pathomorphology of intraperitoneal staphylococcal infection in mice]. / Vliianie rekombinantnogo interleikina-2 na techenie i patomorfologiiu vnutribriushnoi stafilokokkovoi infektsii myshei.

Resistance to the lethal doses of the infectious agent (3 DL100) develops in mice 6 days after the intraperitoneal non-lethal dose of staphylococcus in combination with recombinant interleukin-2 (RIL-2). This effect may be due to the activation of T- and B-dependent zones of the regional lymph nodes and spleen, activation and proliferation of the liver stellate reticulo-endotheliocytes, enhancement of the phagocytic activity of the circulating neutrophil granulocytes. These structural and functional mechanisms may be due to both direct RIL-2 effect in combination with an antigenic stimulation and indirect effect through other interleukins produced by lymphocytes activated by RIL-2.

[Disorders of the immune status in patients with acute diffuse peritonitis]. / Narusheniia immunnogo statusa u bol'nykh ostrym razlitym peritonitom.

Analysis of various parameters of the immune system and non-specific resistance in patients with acute generalized peritonitis (AGP) has indicated that in AGP there is immunodeficiency involving all the links of immune defense and non-specific resistance. The T-cell immunity, primarily T helpers, is afflicted in AGP to the greatest extent. A magnitude of decreases in T helper counts may be used as a prognostic indicator in AGP. The severity of immune defense abnormalities correlates with the duration and intensity of intoxication, there is a "paralysis" of this defense in the terminal stage of AGP.

[Morphogenesis of diabetic microangiopathy]. / Morfogenez diabeticheskoi mikroangiopatii.

Skin biopsies from 40 patients 17 to 75 years old with type I and II diabetes mellitus were studied morphologically. The formation of diabetic microangiopathy starts with the damage of endotheliocytes, vascular permeability disturbance, activation of pericytes and smooth muscle elements with subsequent thickening of basal membranes and capillary and arteriole hyalinosis, these lesions being directly related to the duration of diabetes. Diabetic microangiopathy is a manifestation of the disease and its morphology is similar in both type I and II diabetes.

[Pseudotumors of the orbit and their relationship to granulomatous inflammation]. / Psevdoopukholi orbity i ikh vzaimosviaz' s granulematoznym vospaleniem.

Postoperative material of 225 orbitotomies studied in Helmholtz Research Institute of Eye Diseases for 5 years with the pathohistological diagnosis of chronic inflammation of the orbit tissues was analysed. Granulomatous inflammation was found in 28% of cases and its detailed description in 63 cases is given. Sarcoidosis, Wegener's granulomatosis and other forms of allergic vasculitis and idiopathic forms of "mixed" granulomas are observed around foreign bodies apart from ordinary granulomas. Granulomatous inflammation as a primary manifestation in the orbit of such systemic conditions as Wegener's granulomatosis and sarcoidosis was observed in some cases.

[Chronic interstitial lung diseases. The status of the problem]. / Khronicheskie interstitsial'nye zabolevaniia legkikh. Sostoianie problemy.

Literature and original data on etiology, pathogenesis and morphogenesis of chronic nonspecific lung diseases provided the authors with evidence leading to conclusions on common pathogenesis of the above wide spectrum of the diseases characterized by affected intercellular relations in the lungs. Immunologic homeostasis is suggested as an essential factor resultant in the disease progression along the vicious circle in case of its defective functioning.

[Pathogenesis of diffuse fecal peritonitis]. / O patogeneze razlitogo kalovogo peritonita.

Mongrel albino rats were given intraperitoneal injections of blue pus bacillus endotoxin and human, rat, and guinea-pig fetal suspensions. Analysis of central and peripheral hemodynamics and the indices of the blood coagulation system showed that the severity of the animals' condition was determined by the degree of peripheral hemodynamic disorders associated with changes of the blood rheologic properties. Control of the presence of bacteremia in peritonitis showed that it is encountered both in severe intoxication (injection of human fecal suspension) and in a relatively favourable process (injection of rat fecal suspension). The shifts in the blood coagulation system were characterized by marked individual variability.

[The phenomenon of the accumulation of biological and nonbiological corpuscular particles in mammalian lungs]. / Fenomen nakopleniia v legkikh mlekopitaiushchikh biologicheskikh i nebiologicheskikh korpuskuliarnykh chastits.

When injecting yeast, staphylococcus, polyacrolein microspheres, 3 microns in diameter, into the blood stream of white common rats, it was found that these tend to accumulate in the lumen of pulmonary capillaries, being disposed there in the cytoplasm of macrophages and neutrophils.

[Some problems of the morphogenesis of bullous emphysema of the lungs]. / Nekotorye voprosy morfogeneza bulleznoi émfizemy legkikh.

The analysis of 93 cases of bullous pulmonary emphysema provided evidence for the significance of valvular mechanism of bulla formation operable at two levels: in small bronchi and bronchioles on the one hand and in the area of subpleural fibrosis on the other hand, the latter arising in the result of subpleural atelectasis or subpleural carnificating pneumonia. Relevant pathology is characterized by peripheral acinus situated tangentially to the pleural surface. Electron-microscopic examination revealed pneumocytes of the second type on the internal bulla surface.

[The interrelation of the "local" and the "general" in inflammation]. / Vzaimootnosheniia "mestnogo" i "obshchego" v vospalenii.

The relationships between the local and the general in inflammation are analysed basing on the literature and original data. Local chemoattraction is postulated to be an underlying factor initiating primary local cooperation of cells relevant to inflammation. Being essential in this cooperation, macrophage seems to warrant both the local developments and triggering of general mechanisms of regulation which are relevant to control over subsequent secondary cell cooperation. The latter is biologically aimed at localization of the inflammation focus and separation of its pathogenic factors from intact internal medium. General mechanisms of inflammation control are provided by neuroendocrine, immune, vascular, coagulative, fibrinolytic and other systems, and operate through the products of the acute phase, by immune defence factors and rearrangement of nervous regulation of homeostasis in intact organs and tissues. The result of the regulation manifests with sequential presentation of the inflammation stages in time, correlation of local and general responses intensity. Eventually, local inflammation and lesion involve stress and intoxication which are not considered direct attributes of inflammation, nevertheless can influence general regulatory systems concerned with the course of local inflammation. It is concluded that inflammation implies dialectic unity of local and systemic responses of the body outlined to resolve inflammation and restore homeostasis.