Alternative substrates for conidiogenesis of the entomopathogenic fungus Beauveria bassiana (Bals) Vuillemin (Deuteromycotina: Hyphomycetes) / Substratos alternativos para conidiogênese do fungo entomopatogênico Beauveria bassiana (Bals) Vuillemin (Deuteromycotina: Hyphomycetes)

Abstract Beauveria bassiana is a promising fungus for the biological control of insect pests. The growing costs of conidia production have raised the need to ascertain the efficiency of some low cost substrates. The aim of this study was to analyze the potential use of different raw substrates without nutritional supplement for B. bassiana conidiogenesis. Growth and sporulation were evaluated using 30 g of substrate and 0.3 μL of a conidia suspension (1 x 106 conidia/mL). After 10 days of incubation (70 ± 10% humidity and temperature (T) = 29 ± 1 °C), rice (2.00 x 106 conidia/g substrate), algaroba (2.36 x 106 conidia/g), malt A (1.22 x 106 conidia/g) and malt B (1.75 x 106 conidia/g) showed the highest levels of conidia production. The resulting conidia showed insecticidal activity higher than 80% on coconut termites. These new raw substrates may represent viable alternatives for the production of entomopathogenic fungi for use in the biological control of various insect pests.

Resumo Beauveria bassiana é um fungo promissor no controle biológico de insetos-praga. As crescentes despesas na produção de conídios levantam a necessidade de averiguar a eficiência de alguns substratos de baixo custo. O objetivo deste trabalho foi analisar o potencial de utilização de diferentes substratos brutos para a conidiogênese de B. bassiana. O crescimento e esporulação foram realizados utilizando 30 g do substrato e 0,3 µL da suspensão de conídios (1 x 106 conídios/mL). Após 10 dias de incubação (umidade 70 ± 10% e temperatura T = 29 ± 1° C), o arroz (2,00 x 106 conídios/g de substrato), algaroba (2,36 x 106 conídios/g), malte A (1,22 x 106 conídios/g) e B (1,75 x 106 conídios/g), apresentaram maior produção de conídios. Os conídios produzidos mostraram atividade inseticida sobre o cupim do coqueiro acima de 80% de mortalidade. Estes novos substratos brutos podem representar uma alternativa viável para produção de fungos entomopatogênicos para uso no controle biológico de vários insetos praga.

Alternative substrates for conidiogenesis of the entomopathogenic fungus Beauveria bassiana (Bals) Vuillemin (Deuteromycotina: Hyphomycetes).

Beauveria bassiana is a promising fungus for the biological control of insect pests. The growing costs of conidia production have raised the need to ascertain the efficiency of some low cost substrates. The aim of this study was to analyze the potential use of different raw substrates without nutritional supplement for B. bassiana conidiogenesis. Growth and sporulation were evaluated using 30 g of substrate and 0.3 µL of a conidia suspension (1 x 106 conidia/mL). After 10 days of incubation (70 ± 10% humidity and temperature (T) = 29 ± 1 °C), rice (2.00 x 106 conidia/g substrate), algaroba (2.36 x 106 conidia/g), malt A (1.22 x 106 conidia/g) and malt B (1.75 x 106 conidia/g) showed the highest levels of conidia production. The resulting conidia showed insecticidal activity higher than 80% on coconut termites. These new raw substrates may represent viable alternatives for the production of entomopathogenic fungi for use in the biological control of various insect pests.

Lipolytic Enzymes with Hydrolytic and Esterification Activities Produced by Filamentous Fungi Isolated from Decomposition Leaves in an Aquatic Environment.

Microbial lipases are prominent biocatalysts able to catalyze a wide variety of reactions in aqueous and nonaqueous media. In this work, filamentous fungi isolated from leaves decomposed in an aquatic environment were screened for lipase production with hydrolytic activity and esterification. Agar plates with Tween 20 and Rhodamine B were used for selection, while submerged cultures with olive oil were subsequently used to select 38 filamentous fungi. Trichoderma harzianum, Fusarium solani, Trichoderma harzianum F5, and Penicillium sp. F36 were grown in six different culture media. F. solani presented the highest lipase production (2.37 U/mL) with esterification activity of 0.07 U/mL using medium composed of (g.L-1) KH2PO4 1.00, MgSO4 H2O 1.123, and CuSO4 0.06. Supplementation of this culture medium with organic nitrogen sources increased lipase production by 461.3% using tryptone and by 419.4% using yeast extract. Among the vegetable oils from the Amazon region, degummed cotton oil induced lipase production up to 8.14 U/mL. The lipase produced by F. solani F61 has great potential to application in conventional processes and biodiesel production by transesterification of vegetable oils, as well as food industries in the production of fatty acid esters by hydrolysis and esterification.

Polyphenols journey through blood-brain barrier towards neuronal protection.

Age-related complications such as neurodegenerative disorders are increasing and remain cureless. The possibility of altering the progression or the development of these multifactorial diseases through diet is an emerging and attractive approach with increasing experimental support. We examined the potential of known bioavailable phenolic sulfates, arising from colonic metabolism of berries, to influence hallmarks of neurodegenerative processes. In silico predictions and in vitro transport studies across blood-brain barrier (BBB) endothelial cells, at circulating concentrations, provided evidence for differential transport, likely related to chemical structure. Moreover, endothelial metabolism of these phenolic sulfates produced a plethora of novel chemical entities with further potential bioactivies. Pre-conditioning with phenolic sulfates improved cellular responses to oxidative, excitotoxicity and inflammatory injuries and this attenuation of neuroinflammation was achieved via modulation of NF-κB pathway. Our results support the hypothesis that these small molecules, derived from dietary (poly)phenols may cross the BBB, reach brain cells, modulate microglia-mediated inflammation and exert neuroprotective effects, with potential for alleviation of neurodegenerative diseases.

Vitamin E nanoemulsion activity on stored red blood cells.

BACKGROUND: Stored red blood cells (RBCs) undergo numerous changes that have been termed RBC storage lesion, which can be related to oxidative damage. Vitamin E is an important antioxidant, acting on cell lipids. Thus, this study aimed to investigate vitamin E activity on stored RBCs. METHODS: We prepared a vitamin E nanoemulsion that was added to RBC units and stored at 4 °C. Controls, without vitamin E, were kept under the same conditions. Reactive oxygen species (ROS) production was monitored for up to 35 days of storage. RBC elasticity was also evaluated using an optical tweezer system. RESULTS: Vitamin E-treated samples presented a significant decrease in ROS production. Additionally, the elastic constant for vitamin E-treated RBCs did not differ from the control. CONCLUSION: Vitamin E decreased the amount of ROS in stored RBCs. Because vitamin E acts on lipid oxidation, results suggest that protein oxidation should also be considered a key factor for erythrocyte elastic properties. Thus, further studies combining vitamin E with protein antioxidants deserve attention, aiming to better preserve overall stored RBC properties.

Alternative substrates for conidiogenesis of the entomopathogenic fungus Beauveria bassiana (Bals) Vuillemin (Deuteromycotina: Hyphomycetes)

Abstract Beauveria bassiana is a promising fungus for the biological control of insect pests. The growing costs of conidia production have raised the need to ascertain the efficiency of some low cost substrates. The aim of this study was to analyze the potential use of different raw substrates without nutritional supplement for B. bassiana conidiogenesis. Growth and sporulation were evaluated using 30 g of substrate and 0.3 L of a conidia suspension (1 x 106 conidia/mL). After 10 days of incubation (70 ± 10% humidity and temperature (T) = 29 ± 1 °C), rice (2.00 x 106 conidia/g substrate), algaroba (2.36 x 106 conidia/g), malt A (1.22 x 106 conidia/g) and malt B (1.75 x 106 conidia/g) showed the highest levels of conidia production. The resulting conidia showed insecticidal activity higher than 80% on coconut termites. These new raw substrates may represent viable alternatives for the production of entomopathogenic fungi for use in the biological control of various insect pests.

Resumo Beauveria bassiana é um fungo promissor no controle biológico de insetos-praga. As crescentes despesas na produção de conídios levantam a necessidade de averiguar a eficiência de alguns substratos de baixo custo. O objetivo deste trabalho foi analisar o potencial de utilização de diferentes substratos brutos para a conidiogênese de B. bassiana. O crescimento e esporulação foram realizados utilizando 30 g do substrato e 0,3 µL da suspensão de conídios (1 x 106 conídios/mL). Após 10 dias de incubação (umidade 70 ± 10% e temperatura T = 29 ± 1° C), o arroz (2,00 x 106 conídios/g de substrato), algaroba (2,36 x 106 conídios/g), malte A (1,22 x 106 conídios/g) e B (1,75 x 106 conídios/g), apresentaram maior produção de conídios. Os conídios produzidos mostraram atividade inseticida sobre o cupim do coqueiro acima de 80% de mortalidade. Estes novos substratos brutos podem representar uma alternativa viável para produção de fungos entomopatogênicos para uso no controle biológico de vários insetos praga.

TcCYS4, a cystatin from cocoa, reduces necrosis triggered by MpNEP2 in tobacco plants.

In Brazil, most cocoa bean production occurs in Southern Bahia. Witches' broom disease arrived in this area in 1989 and has since caused heavy losses in production. The disease is caused by the basidiomycete fungus Moniliophthora perniciosa, a hemibiotrophic fungus that produces the necrosis and ethylene-inducting protein (MpNEP2) during infection; this protein can activate cysteine proteases and induce programmed cell death. Cysteine proteases can be modulated by cystatin. In this study, we overexpressed TcCYS4, a cocoa cystatin, in tobacco plants and evaluated the effect on MpNEP2 in model plants. Tccys4 cDNA was cloned into the pCAMBIA 1390 vector and inserted into the tobacco plants via Agrobacterium tumefaciens. Transgene expression was analyzed by reverse transcription-quantitative PCR and Western blot analysis. Transcript and protein levels in Tcccys4:tobacco lines were 8.9- and 1.5-fold higher than in wild-type plants (wt). Tcccys4:tobacco lines showed no change in growth compared to wt plants. CO2 net assimilation (A) increased in Tcccys4:tobacco lines compared to wt plants. Only one line showed statistically significant stomatal conductance (gs) and transpiration rate (E) changes. MpNEP2 was infiltered into the foliar mesophyll of Tcccys4:tobacco lines and wt plants, and necrotic lesions were attenuated in lines highly expressing Tccys4. Our results suggest that cocoa cystatin TcCYS4 affects MpNEP2 activity related to the progression of programmed cell death in tobacco plants. This may occur through the action of cystatin to inhibit cysteine proteases activated by MpNEP2 in plant tissues. Further studies are necessary to examine cystatin in the Theobroma cacao-M. perniciosa pathosystem.

Antioxidant and orofacial anti-nociceptive activities of the stem bark aqueous extract of Anadenanthera colubrina (Velloso) Brenan (Fabaceae).

The anti-nociceptive and antioxidant activities of the Anadenantheracolubrina stem bark aqueous extract (AEAC) were investigated. AEAC (30 µg/mL) reduced 94.8% of 2,2-diphenyl-1-picrylhydrazyl radical and prevented 64% (200 µg/mL) of lipid peroxidation caused by 2,2'-azobis(2-methylpropionamidine) dihydrochloride-induced peroxyl radicals. AEAC treatment (200 and 400 mg/kg) significantly (p < 0.001) reduced mice orofacial nociception in the first (61.4% and 62.6%, respectively) and second (48.9% and 61.9%, respectively) phases of the formalin test. Nociception caused by glutamate was significantly (p < 0.001) reduced by up to 79% at 400 mg/kg, while 56-60% of the nociceptive behaviour induced by capsaicin was significantly inhibited by AEAC (100-400 mg/kg). Mice treated with AEAC did not show changes in motor performance in the Rota-rod apparatus. It appears that AEAC is of pharmacological importance in treating pain due to its anti-nociceptive effects, which were shown to be mediated by central and peripheral mechanisms.

Foxp3 expression and nitric oxide production in peripheral blood mononuclear cells of communicants with pulmonary tuberculosis.

The understanding of the mechanisms involved in the immune response is of significant relevance to the control of tuberculosis (TB), especially in individuals living with patients with TB. To characterize the nitric oxide (NO) production and the Foxp3 marker expression in this population, peripheral blood mononuclear cells of intradomiciliary contacts of individuals with pulmonary tuberculosis with (CTb, susceptible) and without (STb, resistant) previous history of active infection were stimulated in vitro with Mycobacterium tuberculosis antigen (TbAg) and with the mitogen Concanavalin A for 24 and 48 h. The groups analysed did not present significant difference in the Foxp3 mRNA expression nor in the NO production. Negative correlation (P = 0.09) between NO and Foxp3 after a 48-h stimulation with TbAg was observed in the STb group. In this group, after a 24-h culture stimulated with TbAg (P = 0.03), this same correlation was observed. In comparison with the cytokines previously studied by our group (Cavalcanti et al., 2009), a positive correlation was observed between IL-10 and Foxp3 after a 48-h culture of cells from communicants susceptible to tuberculosis (STb) stimulated with TbAg (P = 0.04). Evaluating the entire population, a positive correlation was observed between the cytokine TNF-α and the Foxp3 marker in the cultures stimulated for 24 (P = 0.03) and 48 (P = 0.02) hours with TbAg. Therefore, considering the similarity in the exposure and the individual capacity of responding to the contact with M. tuberculosis, the present study contributes to the comprehension of the immune regulation in individuals living with patients with TB.

American tegumentary leishmaniasis: cytokines and nitric oxide in active disease and after clinical cure, with or without chemotherapy.

The influence of immune response on the treatment of American tegumentary leishmaniasis is pointed by several authors, and the existence of protective immunity in self-healed patients (SH) is also suggested. Thus, interferon-gamma (IFN-γ), tumour necrosis factor-alpha (TNF-α), interleukin (IL-) 10, IL-17, IL-22 and nitric oxide (NO) production was determined in PBMC culture supernatants from patients with active disease (AD) and after therapy, SH patients and healthy subjects, in response to the soluble antigen of Leishmania (Viannia) braziliensis. It was demonstrated that, during the active disease, there is a predominance of IFN-γ and TNF-α, indicating a proinflammatory phase of the response; IL-17 is also highlighted at this clinical state. Also, TNF-α was slightly increased in patients after therapy. NO secretion was noticed in SH individuals, while IL-17 appeared in low levels in these patients and seems to be regulated by NO. The presence of IL-10 was observed in all groups of patients. From this study, we can suggest that in the active disease and after clinical cure, with or without chemotherapy, specific cellular immunity takes part against Leishmania, but with some similarities between the clinical states. Thus, it indicates that the mediators herein described are necessary for the cure to occur.

Antigenic fractions of Leishmania (Viannia) braziliensis: the immune response characterization of patients at the initial phase of disease.

American cutaneous leishmaniasis (ACL) has different clinical manifestations and these manifestations are dependent on the immunological status of the host. As CD4(+) and CD8(+) T cells and their mediators play a fundamental role in the host response to Leishmania and there is also a search for antigenic molecules to be used as future vaccines and tools for prognostic tests, this study characterized ACL patients' immune response after stimulation with soluble and insoluble fractions of L. (V.) braziliensis. We demonstrated a prevailing production of the Th2 cytokines, IL-4 and IL-10 and a specific production of IFN-γ and TNF-α in patients before treatment. There was also a predominance of CD4(+) T cells and a small percentage CD8(+) T cells. The insoluble antigenic fraction primarily stimulated CD4(+) T cells, while the soluble antigenic fraction showed a mixed profile, with CD4(+) T cells being the main responsible for Th2 cytokines and CD8(+) T cells for Th1 cytokines. Therefore, our results showed that a down-modulation of the Th1 type of response occurs in the initial phase of L. braziliensis disease, being the antigenic fractions capable of stimulating a specific immune response.

Immunophenotypic characterization of patients with American cutaneous leishmaniasis prior to and after treatment in Pernambuco, Brazil

Leishmania infections induce a specific activation of host immunological response, particularly characterized by T cell expansion. Studies indicate the importance of the balance between CD4+ and CD8+ T cells, in which the first ones would have their number reduced during the healing process. Meanwhile, CD25+ T cells have been associated with the suppression of the immune response. Since the immune response has an essential role in both healing and progression of diseases, this study aimed to identify the percentage of CD3+, CD4+, CD8+, CD16+ and CD25+ T cells in the peripheral blood of patients afflicted with American cutaneous leishmaniasis (ACL) - before and after treatment - and healthy controls. Peripheral blood was collected and transferred to cytometry tubes containing monoclonal antibodies specific for cell surface markers CD3, CD4, CD8, CD16 e CD25. The immunophenotypic and morphometric parameters of cells were determined by flow cytometry and the results demonstrated a significant increase in the number of T CD8+ cells after treatment, suggesting a cytotoxic T cell response. An increase in CD25+ T cells in patients with active ACL and after treatment was also observed, suggesting the participation of these cells in the modulation of the exacerbated effector response.

Activity and asymmetry index of masticatory muscles in women with and without dysfunction temporomandibular.

PURPOSE: [corrected] Compare the symmetry of the activity and masticatory muscles in individuals with TMD and asymptomatic. METHODS: The study included 50 women, while 31 had temporomandibular disorders (TMD) and 19 were asymptomatic (control group), aged between 19 and 40 years. The volunteers were subjected to clinical examination of the diagnostic criteria in research with TMD (RDC/TMD) with the aim of diagnosing volunteers with or without TMD, and evaluate the electromyographic activity of the right temporalis muscle (TR) and left (TL), right masseter (MR) and left (ML), in situations of rest, isometric contraction of the muscles of the jaw elevators. We obtained the rates of activity and asymmetry in each situation and for the collection and TMD control groups. RESULTS: For the index of activity for the rest there was significant difference (p = 0.0008) between the control group and the TMD group, with predominance of temporal muscle, was not observed difference between groups for the index of activity during the isometric contraction (p = 0.1069). For the index of asymmetry no difference between groups during rest, for the masseter muscles (p = 0.4182) and the temporal (p = 0.7614), and also during the isometry for both masseter muscles (p = 0.8691) and for time (p = 0.6643). CONCLUSIONS: The control group showed prevalence of TMD and temporal muscle during rest, which did not occur in the isometry, and no difference for the index of asymmetry between the groups for the masseter and temporal muscles.

Effect of treatment with HVES on pain and electromyography activity in patients with TMD.

PURPOSE: The aim of this study was to verify the effect of HVES on pain and electromyographic activity of the masticatory muscles in TMD patients, as well as assess the effect of this treatment on the severity of TMD. METHODS: Participated in this study 12 women with TMD that underwent electromyographic exam of the masticatory muscles, and responded to the FAI, RDC/TMD and VAS. RESULTS: After treatment, by the VAS a significant reduction in the intensity of pain it was verified; in the EMG that at rest and during isometric contraction of the mandibular depressor muscles there was a significant reduction in the RMS values; in voluntary contraction during maximal intercuspidation, a significant increase was observed in the values of RMS for the MD and ME muscles. CONCLUSION: HVES reduced the severity of TMD and the intensity of pain, and made the masticatory muscles approach their normal electromyographic pattern.

Utilização de diferentes estimulações elétricas para o tratamento da dor em mulheres com disfunção temporomandibular / Use of different electrical stimulations for treating pain in women with temporomandibular disorders

OBJETIVO: Analisar a intensidade da dor em indivíduos com disfunção temporomandibular (DTM) tratados com dez sessões de estimulação elétrica nervosa transcutânea (TENS) ou estimulação elétrica de Alta Voltagem (EEAV). MÉTODOS: Foram selecionadas 24 mulheres (22,98±1,86 anos) com diagnóstico de DTM, segundo o Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD), sendo 60 por cento com diagnóstico de DTM do grupo Ia e 40 por cento Ia e IIa. As voluntárias foram divididas em dois grupos denominados grupo TENS (GT) e Grupo Alta Voltagem (GAV). Em ambos os grupos as voluntárias receberam dez aplicações da TENS (10Hz modulada em 50 por cento, 200 µs e intensidade no limiar motor) ou da EEAV (10Hz, pulsos gêmeos com 20µs cada e intervalo 100µs interpulsos gêmeos, 100Volts e pólo positivo) duas vezes por semana por 30 minutos. Para mensurar a intensidade da dor, foi utilizada a escala visual analógica (EVA). Para análise estatística, utilizou-se teste t de Student e análise de regressão linear simples. RESULTADOS: Comparando-se as condições pré e pós TENS observa-se uma redução na intensidade da dor (p<0,05) na maioria das sessões, exceto na sexta, sétima e oitava, enquanto a EEAV reduziu a intensidade da dor (p<0,05) em todas as sessões. Avaliando-se os valores pré-aplicação, os dois recursos diminuíram a intensidade de dor de forma uniforme ao longo das dez sessões (p<0,05). CONCLUSÕES: A TENS e a EEAV promoveram redução da intensidade da dor em mulheres com DTM, sendo a EEAV mais um recurso indicado para o tratamento desses pacientes.

OBJECTIVE: To analyze pain intensity in individuals with temporomandibular disorder (TMD) who were treated with ten sessions of transcutaneous electrical nerve stimulation (TENS) or high voltage electrical stimulation (HVES). METHODS: Twenty-four women (22.98±1.86 years old) with a diagnosis of TMD in accordance with the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) were selected. 60 percent of the subjects had a diagnosis of TMD classified as group Ia and 40 percent as Ia and IIa. They were divided into two groups named the TENS group (TG) and the high voltage group (HVG). Each individual received ten applications of either TENS (10Hz, modulated at 50 percent, 200µs and motor threshold intensity) or HVES (10Hz, twin pulses of 20µs each at intervals of 100µs between the twin pulses, 100volts and positive pole) twice a week for 30 minutes. To measure the pain intensity, a visual analog scale (VAS) was used. Statistical analyses were performed using Student's t test and simple linear regression. RESULTS: Comparison of the pre and post-TENS conditions showed diminished pain intensity (p<0.05) at most sessions except for sessions 6, 7 and 8. In contrast, HVES reduced the pain intensity at all sessions (p<0.05). Evaluation of the pre-application values showed that both treatments decreased the pain intensity uniformly over the ten sessions (p<0.05). CONCLUSIONS: TENS and HVES both promoted reductions in pain intensity in women with TMD. HVES is a therapeutic resource recommended for such patients.

Mechanism and treatment of sulfide-induced coma: a rat model.

Sodium hydrosulfide and dimethylsulfide duplicate the effects of hydrogen sulfide in causing coma in Sprague-Dawley rats and are additive for lethality. Nitrite, pyruvate and dithiothreitol had no significant effect on coma or lethality but bicarbonate with and without glucose reduced duration of coma. This finding suggests an antidotal treatment.

Efeitos do Método Mãe Canguru nos sinais vitais de recém-nascidos pré-termo de baixo peso / Effects of Kangaroo Mother Care on the vital signs of low-weight preterm newborns

OBJETIVO: Avaliar as freqüências cardíaca e respiratória, a pressão arterial média, a temperatura e a saturação periférica de oxigênio dos recém-nascidos pré-termo (RNPT) de baixo peso, antes e após a aplicação do MMC. MÉTODOS: Foram avaliados 22 RNPT de baixo peso, saudáveis, de ambos os sexos, não portadores de deficiências neurológicas, cardíacas e/ou respiratórias. A avaliação foi realizada após trinta minutos de permanência do RNPT em berço comum e após trinta minutos de aplicação do MMC, por 3 dias consecutivos. Para a avaliação, foram utilizados monitor cardíaco com dispositivo para medida da pressão arterial média de forma não invasiva e sensor para a oximetria de pulso, termômetro e cronômetro. RESULTADOS: Os resultados não mostraram alterações significativas quanto à pressão arterial média (p> 0,05) e freqüência cardíaca (p> 0,05) após a aplicação do MMC, mas, por outro lado, houve aumento significativo da temperatura axilar (p< 0,05) e da saturação periférica de oxigênio (p< 0,05) e diminuição significativa da freqüência respiratória (p< 0,05). CONCLUSÃO: O MMC promove a melhora da temperatura corporal, o aumento da saturação periférica de oxigênio, melhorando a oxigenação tecidual, e a redução na freqüência respiratória, trazendo maior conforto respiratório aos RNPT, sugerindo que o MMC contribui para alterações benéficas nos sinais vitais dos RNPT.

OBJECTIVE: The aim of this study was to evaluate the heart and respiration rates, mean arterial pressure, temperature and peripheral oxygen saturation of low-weight preterm newborns, before and after the application of kangaroo mother care. METHOD: Twenty-two healthy low-weight preterm newborns of both sexes were studied. None of them had neurological, cardiac and/or respiratory deficiencies. Assessments were made after the newborn had been left in an ordinary cot for 30 minutes and after 30 minutes of kangaroo mother care, on three consecutive days. For these evaluations, a heart monitor with a device for non-invasively measuring mean arterial pressure, a sensor for pulse oximetry, a thermometer and a chronometer were utilized. RESULTS: There were no significant changes in mean arterial pressure (p> 0.05) or heart rate (p> 0.05) after applying kangaroo mother care. However, there were significant increases in axillary temperature (p< 0.05) and peripheral oxygen saturation (p< 0.05), and a significant decrease in respiration rate (p< 0.05). CONCLUSION: Kangaroo mother care promotes improvement in body temperature, increased peripheral oxygen saturation (thus improving tissue oxygenation), and decreased respiration rate (thus providing greater respiratory comfort for the newborns). This suggests that kangaroo mother care contributes towards beneficial alterations in the low-weight newborns' vital signs.

Use of immunofluorescence and confocal laser scanning microscopy in identifying rare cases of anti-neutrophil cytoplasmic antibodies (ANCA) showing dual specificities to myeloperoxidase and proteinase3.

Anti-neutrophil cytoplasmic antibodies (ANCA) are the immunodiagnostic markers for idiopathic necrotizing crescentic glomerulonephritis affecting mainly medium to small sized blood vessels. The diagnosis of ANCA associated vasculitis (AAV) is mainly based on clinical and histopathological characteristics along with the serological evidence. Immunofluorescence microscopy (IIF) is considered as the "gold standard" for ANCA detection, and ANCA showing two major patterns ie, cytoplasmic (c-ANCA) and perinuclear (p-ANCA) react with different antigenic targets of neutrophils like Proteinase3 (PR3) and Myeloperoxidase (MPO). A third unusual and rare immunofluorescence pattern called as "X- ANCA" or atypical ANCA is also sometimes seen. The difficulty in identification of ANCA immunofluorescence patterns is mainly seen due to the rare dual patterns seen in the same sera and also the additional nuclear immunofluorescence seen due to presence of anti-nuclear antibodies. ANCA testing by immunofluorescence and Confocal Laser scanning microscopy, as well as by specific ELISAs for detection of anti-PR3 and anti-MPO antibodies have helped in improving the diagnosis. Patients having dual specificities to MPO and PR3 in a patient is a rare finding. Among 425 clinically and histopathologically proven cases of AAV, eight patients (1.9%) had dual specificities, of which five patients showed mixed immunofluorescence pattern and 3 patients showed X-ANCA pattern which was confirmed by both immunofluorescence and Confocal Laser scanning microscopy and the dual specificities to MPO and PR3 were detected by individual ELISAs.

Anti-neutrophil cytoplasmic antibodies (ANCA) in systemic lupus erythematosus: prevalence, clinical associations and correlation with other autoantibodies.

AIM: This study was undertaken to clarify the nature of anti-neutrophil cytoplasmic antibodies (ANCA) along with other autoantibodies in lupus nephritis (LN) patients and in systemic lupus erythematosus (SLE) patients without nephritis and to know their correlation with clinical manifestations and presence of other autoantibodies. MATERIAL AND METHODS: Fourty one LN patients and 18 SLE patients without nephritis were studied. LN patients were subdivided into diffuse proliferative glomerulonephritis (DPGN), focal proliferative glomerulonephritis (FPGN), rapidly progressive glomerulonephritis (RPGN) and membranoproliferative glomerulonephritis (MPGN). Anti-neutrophil cytoplasmic antibodies (ANCA) were detected by indirect immunofluorescence and confocal laser scanning microscope using PMN and HL60 cells. ANCA specificities like anti-myeloperoxidase (anti-MPO), anti-proteinase 3 (anti-PR3), anti-lactoferrin (anti-LF) and anti-cathepsin G (anti-CG) were detected by ELISA. Other autoantibodies like anti-nuclear antibodies (ANA), anti-double stranded DNA (anti-dsDNA), anti-single stranded DNA(anti-ssDNA), anti-ribonucleoproteins (anti-nRNP), anti-Smith antibodies (anti-Sm) and rheumatoid factor (RF) were also tested. RESULTS: ANCA was detected in 37.3% patients. The predominant ANCA pattern was perinuclear (p-ANCA). ANCA positivity was higher in LN patients and when confirmed by ELISA, 54.5% ANCA positives had anti-myeloperoxidase (anti-MPO). The cytoplasmic ANCA (c-ANCA) pattern was not seen in any patient. Two patients having FPGN with crescents showed atypical 'X-ANCA' pattern with dual specificity to anti-MPO and anti-PR3 by ELISA. The titers of ANCA were more in LN as compared to SLE without nephritis. LN cases having DPGN, FPGN, RPGN with crescents had higher titer p-ANCA positivity with corresponding anti-MPO antibodies, along with ANA, anti-dsDNA, anti-ssDNA and anti-Sm + anti-nRNP and also high SLEDAI scores. CONCLUSION: ANCA in SLE may be used as a serological marker along with clinical and histopathological assessment to differentiate vasculitides in LN cases from SLE without nephritis.