Nonfunctional overreaching leads to inflammation and myostatin upregulation in swiss mice.

The aims of the this study were a) to verify whether the performance decrease induced by nonfunctional overreaching (NFOR) is linked to high concentrations of cytokines in serum, skeletal muscles and liver; b) to verify muscle myostatin adaptation to NFOR; c) to verify the effects of chronic glucose supplementation on the parameters mentioned above. Mice were divided into control (C), trained (TR), overtrained (OTR) and supplemented overtrained (OTR + S). The incremental load test (ILT) and exhaustive test (ET) were used to measure performances before and after exercise protocols. 24 h after ET, muscles and liver were removed and stored at -80°C for subsequent measurements. Total blood was collected from decapitation for subsequent determination of cytokine concentrations. Generally, OTR and OTR + S presented higher contents of IL-6, TNF-alpha, GLUT-4 and myostatin in muscle samples compared to C and TR. Glucose supplementation attenuated the high contents of IL-6, TNF-alpha and IL-15 in liver, and of IL-6 in serum. In summary, NFOR led to low-grade chronic inflammation and myostatin upregulation.

Effects of chronic ethanol consumption in experimental sepsis.

AIMS: To evaluate the effects of chronic ethanol consumption on the development and the pathophysiology of sepsis, using an experimental model of polymicrobial peritonitis by feces i.p. injection. METHODS: Forty-day-old male Wistar rats were divided into groups for two experiments: A and B. Experiment A was performed for determination of mortality rates, while experiment B was designed for biochemical analysis and measurement of cytokines before and after sepsis. In both the experiments, treated animals were exposed to a 10% ethanol solution as the single drinking source for 4 weeks, while untreated animals were exposed to tap water over the same period. Food was provided ad libitum. After this period, the animals underwent i.p. fecal injection for induction of sepsis. RESULTS: Experiment A showed that higher doses of ethanol resulted in early mortality from sepsis that was correlated with the alcohol consumption (high dose = 85.7%, low dose = 14.3%, P = 0.027). In experiment B, cytokine analysis demonstrated important changes resulting from sepsis, which were further affected by ethanol exposure. In addition, glucose and creatinine levels decreased and increased, respectively, after sepsis, but a significant change occurred only in the ethanol group (P < 0.003 glucose, P < 0.01 creatinine). The levels of pro-inflammatory cytokines, interleukin-6 and tumor necrosis factor-α, increased after sepsis, but were less evident after ethanol exposure. CONCLUSION: These differences may be the result of either early mortality or an increase in the severity of the septic process. Taking into account the high mortality rate and the extreme severity of sepsis after alcohol consumption, often encouraged by advertising, a caution should be given to patients with severe infections and a history of alcohol abuse.

Binge drinking and insomnia in students from health sciences at one university in Rio de Janeiro, Brazil.

In spite of the many studies examining alcohol consumption, recent reviews have indicated that binge drinking has not been extensively studied. Furthermore, it is becoming increasingly clear that sleep is associated with many physiological functions and to drug addictions. The present study aimed to evaluate the relationship between alcohol binge drinking and insomnia in college students of health sciences. All first-year health sciences students (n=286) were evaluated in a cross-sectional study. Envelopes containing the Insomnia Severity Index (ISI), the Alcohol, Smoking, and Substance Involvement Screening Test (ASSIST), and questions capturing sociodemographic data were distributed and collected in classes. It was found that most non-drinkers were female (70.6%), although there were no sex-related differences in the number of binge drinkers (more than 5 drinks on each occasion at least once a week), allowing statistical comparison. The Mann-Whitney U test indicated that the ISI scores were significantly greater in female than male binge drinkers (P=0.014). Moderate or severe insomnia was reported by 23% of the sample, with alcohol being the most frequently associated substance. A specialized intervention was suggested by ASSIST: brief for marijuana (19.2%) and tobacco (23.3%) use, and moderate (31.5%) or intensive (1.4%) for alcohol consumers. The data highlighted the need to pay attention to the habits of college students beyond obtaining scientific information. New data suggesting the influence of genetics on insomnia may be of importance when performing additional studies on the sex differences in alcohol binge drinking.

Risk factors and characteristics of ocular complications, and efficacy of autologous serum tears after haematopoietic progenitor cell transplantation.

The objective of the study was to evaluate the frequency and clinical characteristics of ocular complications and their risk factors, as well as autologous serum tears (AST) for the treatment of dry eye in these patients. Data from the files of 124 patients who had undergone allogeneic haematopoietic progenitor cell transplantation (HPCT) were evaluated. In addition, 33 HPCT patients were examined and their data were compared with controls. Analysis of tears and AST was performed. Dry eye manifestation occurred in 32% of patients and was positively correlated with age over 27 years (P = 0.05), peripheral blood progenitor cell transplant (P = 0.002), chronic graft-versus-host disease (P = 0.0027), and chronic or acute myeloid leukaemia (P = 0.001). Dry mouth and Schirmer test < 5 mm were predictive factors for dry eye in HPCT patients (P = 0.002 and odds ratio 3.9 and P = 0.007, odds ratio = 5.9, respectively). Microbiological analysis revealed that six of 11 AST samples were contaminated after 30 days of use. The present study supports the role of potential risk factors for ocular complications and key elements to detect alterations in the tear film from HPCT patients. In addition, AST contamination must be considered after longer periods of use.

Fibroblast Biomarkers of Sporadic Parkinson's Disease and LRRK2 Kinase Inhibition.

It has been uncertain whether specific disease-relevant biomarker phenotypes can be found using sporadic Parkinson's disease (PD) patient-derived samples, as it has been proposed that there may be a plethora of underlying causes and pathological mechanisms. Fibroblasts derived from familial PD patients harboring leucine-rich repeat kinase 2 (LRRK2), PTEN-induced putative kinase 1 (PINK1), and Parkin mutations show clear disease-relevant mitochondrial phenotypes, which are exacerbated under conditions of pharmacological stress. We utilized fibroblasts derived from non-familial sporadic PD patients (without LRRK2 mutations) or LRRK2 mutation carriers to directly compare the cellular phenotypes during and after mitochondrial stress. We then determined the effects of pharmacological LRRK2 kinase inhibition using LRRK2-in-1. We found that there were two distinct populations of sporadic PD patient-derived fibroblast lines. One group of sporadic PD lines was highly susceptible to valinomycin-induced mitochondrial depolarization, emulating the mutant LRRK2 phenotype. These lines showed elevated mitochondrial superoxide/ nitric oxide levels, displayed increased mitochondrial and lysosome co-localization, and an increased rate of mitochondrial collapse, which corresponded with changes in mitochondrial fission and fusion proteins. The application of LRRK2-in-1 reversed decreased levels of mitochondrial and lysosome co-localization and partially restored mitochondrial network associated proteins and the mitochondrial membrane potential in the fibroblasts. This study identifies novel mitochondrial biomarkers in sporadic PD patient-derived fibroblast lines, which could be used as preclinical tools in which to test novel and known neuroprotective compounds.

Binge drinking and insomnia in students from health sciences at one university in Rio de Janeiro, Brazil

In spite of the many studies examining alcohol consumption, recent reviews have indicated that binge drinking has not been extensively studied. Furthermore, it is becoming increasingly clear that sleep is associated with many physiological functions and to drug addictions. The present study aimed to evaluate the relationship between alcohol binge drinking and insomnia in college students of health sciences. All first-year health sciences students (n=286) were evaluated in a cross-sectional study. Envelopes containing the Insomnia Severity Index (ISI), the Alcohol, Smoking, and Substance Involvement Screening Test (ASSIST), and questions capturing sociodemographic data were distributed and collected in classes. It was found that most non-drinkers were female (70.6%), although there were no sex-related differences in the number of binge drinkers (more than 5 drinks on each occasion at least once a week), allowing statistical comparison. The Mann-Whitney U test indicated that the ISI scores were significantly greater in female than male binge drinkers (P=0.014). Moderate or severe insomnia was reported by 23% of the sample, with alcohol being the most frequently associated substance. A specialized intervention was suggested by ASSIST: brief for marijuana (19.2%) and tobacco (23.3%) use, and moderate (31.5%) or intensive (1.4%) for alcohol consumers. The data highlighted the need to pay attention to the habits of college students beyond obtaining scientific information. New data suggesting the influence of genetics on insomnia may be of importance when performing additional studies on the sex differences in alcohol binge drinking.

Androgen influence on the meibomian gland.

PURPOSE: The hypothesis in the study was that androgens control meibomian gland function, regulate the quality and/or quantity of lipids produced by this tissue, and promote the formation of the tear film's lipid layer. To test this hypothesis, a study was conducted to determine whether androgen receptor protein exists in the epithelial cell nuclei of rat meibomian glands and, in addition, whether androgen deficiency and/or treatment influences the gross morphology, neutral lipid content, and fatty acid profile of the rabbit meibomian gland, as well as the appearance of the tear film lipid layer. METHODS: Rat lids were obtained and processed for immunohistochemistry. Meibomian glands from intact, androgen- and/or placebo-treated rabbits were analyzed by histology, and glandular lipids were evaluated by gas chromatography, high-performance liquid chromatography (HPLC), and mass spectrometry. The rabbit tear film lipid layer was assessed by interferometry. RESULTS: In the current study androgen receptor protein existed within acinar epithelial cell nuclei of rat meibomian glands; androgen deficiency was associated with alterations in the lipid content of the rabbit meibomian gland; 19-nortestosterone treatment modulated the fatty acid profile in the total and neutral lipid fractions of the rabbit meibomian gland; and androgens did not appear to influence the gross morphology of meibomian tissue or to exert a demonstrable effect on the rabbit tear film lipid layer. CONCLUSIONS: The findings show that the meibomian gland is an androgen target organ and that androgens influence the lipid profile within this tissue. However, the extent to which androgens regulate the production of these lipids and whether this action may impact tear film stability remain to be determined.

Insulin-induced tyrosine phosphorylation of Shc in liver, muscle and adipose tissue of insulin resistant rats.

Insulin stimulates rapid tyrosine phosphorylation of the protein Shc, which subsequently binds to Grb2, resulting in the activation of a complex mitogenic signaling network. In this study, we examined the levels of Shc protein, its phosphorylation state and Shc-Grb2 association in liver, muscle and adipose tissue before and after insulin administration in three animal models of insulin resistance (chronic dexamethasone treatment, 72-h starvation and aging). There were no differences in Shc protein expression between tissues from control and insulin resistant animals. In fasted hypoinsulinemic rats, there was a decrease in insulin-induced Shc phosphorylation in liver and adipose tissue. However, a significant increase in Shc phosphorylation was observed in liver and muscle from dexamethasone-treated hyperinsulinemic rats and in liver, muscle and adipose tissue of hyperinsulinemic 20-month-old rats. Alterations in Shc phosphorylation correlated well with the level of Shc-Grb2 association. These results indicate that Shc tyrosyl phosphorylation and Shc-Grb2 association are regulated in the different types of insulin resistance and that this regulation is apparently related to the animals' plasma insulin levels. The Shc-Grb2 association is directly related to the insulin-induced tyrosyl phosphorylation of Shc.

GVHD dry eyes treated with autologous serum tears.

Two cases of GVHD with severe dry eyes are reported where conventional therapy failed to control ocular signs and symptoms. Autologous serum tears, however, resulted in a beneficial clinical effect with marked attenuation of the symptoms. This therapy proved to be safe during 10 months of treatment. Bone Marrow Transplantation (2000).

Androgens and dry eye in Sjögren's syndrome.

Sjögren's syndrome is an extremely complex and currently incurable autoimmune disorder, which occurs primarily in females, and is associated with lacrimal gland inflammation, meibomian gland dysfunction, and severe dry eye. We hypothesize that androgen deficiency, which reportedly occurs in primary and secondary Sjögren's syndrome (e.g., systemic lupus erythematosus, rheumatoid arthritis), is a critical etiologic factor in the pathogenesis of dry eye syndromes. We further hypothesize that androgen treatment to the ocular surface will promote both lacrimal and meibomian gland function and alleviate both "aqueous-deficient" and "evaporative" dry eye. Our results demonstrate that androgens regulate both lacrimal and meibomian gland function, and suggest that topical androgen administration may serve as a safe and effective therapy for the treatment of dry eye in Sjögren's syndrome.

Bancroftian filariasis in Maceio, state of Alagoas, Brazil: Observations on Culex quinquefasciatus after blood feeding on individuals with different densities of microfilariae in the peripheral blood stream.

Females of laboratory-reared Culex quinquefasciatus were allowed to take blood meals on individuals with low (1-100 microfilariae/ml of peripheral blood), moderate (101-500 microfilariae/ml), and high (> 500 microfilariae/ml) microfilaremia. The mosquitoes ingested 1.39-3.80 microl of blood and infective third-stage (L3) larvae were first recorded 13-14 days after the infecting blood meal. The number of microfilariae ingested by mosquitoes was proportional to the density of microfilariae in the peripheral blood of the human subjects, but with a concentration factor of up to 6.5 times the expected number. Survival of mosquitoes was not influenced by the density of microfilariae in the peripheral blood of infected individuals. Infectivity indices were proportional to microfilaremia in human subjects. The number of L3 larvae/female (intensity of infection) was not influenced by individual microfilaremia. The highest vector efficiency and the best experimental infection index were recorded in mosquitoes that blood fed on individuals with moderate microfilaremia. The results are discussed in relation to the transmission on Bancroftian filariasis in the study area.

Identification of androgen receptor protein and 5alpha-reductase mRNA in human ocular tissues.

BACKGROUND/AIMS: Androgens have been reported to influence the structural organisation, functional activity, and/or pathological features of many ocular tissues. In addition, these hormones have been proposed as a topical therapy for such conditions as dry eye syndromes, corneal wound healing, and high intraocular pressure. To advance our understanding of androgen action in the eye, the purpose of the present study was twofold: firstly, to determine whether tissues of the anterior and posterior segments contain androgen receptor protein, which might make them susceptible to hormone effects following topical application; and, secondly, to examine whether these tissues contain the mRNA for types 1 and/or 2 5alpha-reductase, an enzyme that converts testosterone to the very potent metabolite, dihydrotestosterone. METHODS: Human ocular tissues and cells were obtained and processed for histochemical and molecular biological procedures. Androgen receptor protein was identified by utilising specific immunoperoxidase techniques. The analysis of type 1 and type 2 5alpha-reductase mRNAs was performed by the use of RT-PCR, agarose gel electrophoresis, and DNA sequence analysis. All immunohistochemical evaluations and PCR amplifications included positive and negative controls. RESULTS: These findings show that androgen receptor protein exists in the human lacrimal gland, meibomian gland, cornea, bulbar and forniceal conjunctivae, lens epithelial cells, and retinal pigment epithelial cells. In addition, our results demonstrate that the mRNAs for types 1 and 2 5alpha-reductase occur in the human lacrimal gland, meibomian gland, bulbar conjunctiva, cornea, and RPE cells. CONCLUSION: These combined results indicate that multiple ocular tissues may be target sites for androgen action.

Characterization of the insulin-signaling pathway in lacrimal and salivary glands of rats.

PURPOSE: Insulin has been acknowledged as a mediator of several physiological events in lacrimal and salivary glands. We investigated the presence of insulin receptors and of insulin-induced autophosphorylation of the insulin receptor and activation of elements involved in the early steps of insulin signaling in lacrimal and salivary glands of rats. METHODS: Lacrimal and salivary glands of Wistar rats were removed and processed for immunohistochemistry using anti-insulin receptor and anti-IGF-1 receptor antibodies. The activation of insulin receptors following insulin treatment, and the involvement of insulin receptor substrates-1 and -2, Shc, JAK-2 and STAT-1, were analyzed by immunoprecipitation, followed by SDS-PAGE and immunoblotting of rat lacrimal and salivary glands after exposure to insulin. RESULTS: Insulin and IGF-1 receptors were present in rat lacrimal and salivary glands and were located predominantly in the cytoplasm and plasma membrane. Functional studies demonstrated that insulin induced a dose-dependent phosphorylation of the insulin receptor, IGF-1R, insulin receptor substrates-1 and -2, Shc, and STAT-1. In rats with streptozotocin-induced diabetes mellitus there was a significant reduction in insulin-induced insulin receptor and STAT-1 phosphorylation in the lacrimal gland but not in the salivary gland; there was no influence on Shc phosphorylation in either tissue. CONCLUSIONS: The present results indicate that insulin and IGF-1 receptors are expressed in lacrimal and salivary glands, and that insulin can induce the phosphorylation of its receptor and activate elements involved in the early steps of insulin signaling in both tissues.

In vitro activity of natural and synthetic naphthoquinones against erythrocytic stages of Plasmodium falciparum.

In an attempt to identify new antimalarial compounds we studied the blood schizonticide effect of chemically defined natural products which were isolated from plants (Bignoniaceae) or synthesized. Different concentrations of these drugs (up to 20 microM) were incubated in vitro with blood forms of Plasmodium falciparum for 72 h. A total of 12 drugs of the naphthoquinones group were tested. Eight of them were isolated from plants (NP) and 4 synthesized (S). Three of the drugs (2 NP and one S) were very active and completely inhibited parasite growth at the higher concentrations used (20 microM); 5 drugs were partially active (3S and 2NP) and 3 (NP) were totally inactive. Lapachol was among the drugs tested. Although it has been considered a potential antimalarial agent, it exhibited very low activity (20% inhibition of schizogony). The antimalarial activity of our naphthoquinones against drug-resistant strains was superior to that of chloroquine and quinine which were used as controls. Two of the P. falciparum strains used for the tests were strongly chloroquine resistant. If these naphthoquinones prove to be active in vivo and are of low toxicity, they will be promising candidates for treatment of human malaria particularly since they are easily synthesized.

The immunodominant surface antigen of Plasmodium gallinaceum is present in both the salivary gland and oocyst sporozoites.

1. Monoclonal antibodies (MAbs) against surface antigens of Plasmodium gallinaceum sporozoites, an avian malaria parasite, were produced using spleen cells from mice immunized with sporozoites from mosquito salivary glands (SGS) or from midguts containing oocysts (OoS). 2. All of the 15 MAbs tested (11 anti-SGS and 4 anti-OoS) reacted with SGS and OoS by indirect immunofluorescence and circumsporozoite precipitation reactions. Fourteen of these MAbs (11 anti-SGS and 3 anti-OoS) produced a Western blot (WB) pattern identical to that produced with serum from mice hyperimmunized with viable intact sporozoites. 3. All MAbs and the immune sera recognized only two polypeptide bands of approximate molecular weight 76 and 64 kDa. 4. No difference in the WB pattern was observed when 9- or 12-day SGS or OoS extracts were used as antigens in WB. This antigenic similarity was confirmed when the total protein extracts were visualized on silver-stained SDS-PAGE gel.

Refractoriness of Callithrix penicillata red blood cells to Plasmodium falciparum in vivo and in vitro.

Attempts to infect the New World marmot Callithrix penicillata with Plasmodium falciparum were unsuccessful. Attempts were also made to infect red blood cells of C. penicillata and Saimiri sciureus with P. falciparum in vitro, and these too were unsuccessful due to a high rate of hemolysis produced by apparently adverse culture conditions. It is concluded that modifications to the existing culture conditions will need to be made before successful parasitemia can be induced in vitro in simian erythrocytes.

Survey of medicinal plants used as antimalarials in the Amazon.

Plants traditionally employed for the treatment of malaria in certain areas of Brazil, where this disease is prevalent, were surveyed by interviewing natives and migrants in the Amazon Region. Forty-one plants used for malarial treatment and/or for the related symptoms (fever and liver disorders) were collected and identified. Given the potential of Brazil's forests and medicinal plants, research on traditional plant-based remedies in this country may lead to the development of new drugs.

Experimental infections of simians with human malaria: attempts to infect Callithrix penicillata with Plasmodium falciparum.

After reviewing the use of non-human primates of the Old and New Worlds for human malaria research, we concluded that another experimental animal which is easily available to use and possible to rear indoors is needed. Thus, we studied the susceptibility of the marmoset Callithrix penicillata to Plasmodium falciparum erythrocytic infections. The marmosets received various P. falciparum human isolates, directly from a patient and from continuous cultures. The Palo Alto strain, which has been adapted to the night monkey Aotus trivirgatus and further maintained in the squirrel monkey Saimiri sciureus was also used. In a total of 20 marmosets we performed 31 inoculations, with 10(5) to 10(9) parasites, intraperitoneally, intracardiacly or intravenously. Blood samples from each animal were examined daily up to day 90 post-inoculation. None of the intact marmosets developed patent infections. Four out of 19 C. penicillata, previously splenectomized, showed circulating parasites for up to five days after intravenous inoculation with the Palo Alto strain, becoming negative thereafter. Neither the addition to the simian diet of p-aminobenzoic acid, essential for the parasite metabolism, nor drug-immunosuppression, improved the marmoset susceptibility to P. falciparum.

[Bancroftian filariasis in urban areas of Alagoas State, Northeast Brazil: study in the general population]. / Filariose bancroftiana em áreas urbanas do Estado de Alagoas, nordeste do Brasil: estudo em populaçao geral.

In order to assess the present status of lymphatic filariasis in Alagoas State, Brazil, hemoscopic surveys were carried out in the human population of the three different physiographic regions of the State. Blood samples were collected by thick smear technique taken after 10:00 p.m. From a total of 101 cities of the State, 10 were randomly selected, Bancroftian filariasis was only found in Maceió, the capital of Alagoas State. In a cross-sectional survey conducted among the general population of 4 neighborhoods in the city, 10,973 individuals were examined, 226 were microfilaraemic with prevalence in the city neighborhoods ranging from 0 to 5.4%. Prevalence rates and microfilariae density were significantly higher in males. Among the examined inhabitants born out of Maceió, microfilariae carriers had lived a significantly longer time in endemic areas than the amicrofilaraemic subjects. Based on the data obtained in the present study, measures to avoid expansion of bancroftian filariasis in the region were established.

[Present status of bancroftian filariasis in Maceió, State of Alagoas, Brazil]. / Situação atual da filariose bancroftiana na cidade de Maceió, estado de Alagoas, Brasil.

Epidemiological and entomological surveys were carried out in the human and mosquito populations in Maceió, Alagoas, in order to assess the present status of bancroftian lymphatic filariasis. Examination of thick blood smears of 10,450 students from different areas of the city revealed 0.66% Wuchereria bancrofti microfilaria carriers. The distribution of filariasis is focal in the city, 80% of the individuals with patent infection living in two neighboring areas with 1.24% and 5.25% prevalence. Parallel studies performed with samples of all age groups in the human population showed similar microfilaria prevalence rates observed previously in the student survey. However, thick blood smears taken from members of families with at least one subject with patent infection gave a prevalence six times greater suggesting, increased transmission in households. The percentage of carriers was higher in the youngest age group (< 20 years). Culex quinquefasciatus mosquitos caught at the locations where the autochthonous cases were found presented natural infection rates ranging from 0.28% to 4.62%. The combination of all these findings indicates occurrence of active transmission of W. bancrofti in the urban area of Maceió, Alagoas State. Based on these data, measures for the potential control of filariasis were planned.