Molecular docking studies and evaluation of the antiretroviral activity and cytotoxicity of the species Lafoensia pacari Saint-Hilaire.

Interest in antiviral plant species has grown exponentially and some have been reported to have anti-HIV properties. This research aims to perform the bio-guided phytochemical fractionation by antiretroviral activity of Lafoensia pacari stem barks. This in vitro experimental study involved the preparation of plant material, obtention of ethanolic extract, fractionation, purification, identification and quantification of fractions, acid-base extraction, nuclear magnetic resonance, HIV-1 RT inhibition test and molecular docking studies. From the bio-guided fractionation by the antiretroviral activity there was a higher activity in the acetanolic subfractions, highlighting the acetate subfraction - neutrals with 60.98% of RT inhibition and ellagic acid with 88.61% of RT inhibition and absence of cytotoxicity. The macrophage lineage cytotoxicity assay showed that the chloroform fraction was more toxic than the acetate fraction. The analysis of the J-resolved spectrum in the aromatic region showed a singlet at 7.48 and 6.93 ppm which was identified as ellagic acid and gallic acid, respectively. The 5TIQ enzyme obtained better affinity parameter with the ellagic acid ligand, which was confirmed by the HSQC-1H-13C spectra. Gallic acid was also favorable to form interaction with the 5TIQ enzyme, being confirmed through the HSQC-1H-13C spectrum. From the PreADMET evaluation it was found that ellagic acid is a promising molecule for its RT inhibition activity and pharmacokinetic and toxicity parameters.

Monitoring bovine tick fever on a dairy farm: An economic proposal for rational use of medications.

In this study, we evaluated the monitoring of tick fever (TF) in a Brazilian dairy farm in the Minas Gerais state, Brazil, from July 10 to August 4, 2018. We aimed to identify diagnostic and treatment flaws in the protocol adopted by the farm, and to establish a novel and accurate TF monitoring protocol based on precision dairy farming and rational use of antimicrobials and antiparasitic drugs, while evaluating the economic benefits of the proposed strategy. We monitored TF in 395 heifer calves aged between 3 and 14 mo. According to the farm's standard protocol, all calves with an increase of 0.5°C in rectal temperature compared with the previous week's measurement were treated for Anaplasma spp. and Babesia spp. Blood smears were collected from the tail tip of the treated calves. During the last week of the study, we prepared blood smears of all calves regardless of treatment indication. Economic analysis was performed. The results indicated that at least 56.86% (261/459) of the calves did not require treatment for TF, whereas only 23.09% (106/459) had treatment indications. Negative blood smears (45.97%; 211/459) indicated the possibility of calves being affected by another disease or a condition that was not being adequately treated or those not necessarily sick. These results demonstrate the excessive use of medications, representing a direct economic loss, in addition to potentially favoring the occurrence of resistance to antimicrobials. In contrast, 9.42% (26/276) of calves had no treatment indication based on rectal temperature but had treatment indications based on blood smears. Only 5.73% (42/735) of blood smears had co-infection with hemopathogens, and none had triple co-infection. Therefore, we proposed the monitoring of TF using rectal temperature and microscopic analysis. If implemented, this strategy would result in a direct annual savings of approximately $22,638.96 (77.99%) related to medication for the treatment of TF. Therefore, implementing the proposed protocol would be cheaper than treatment based only on rectal temperatures. The currently implemented TF protocols overestimate the occurrence of TF, resulting in overtreatment. Thus, implementing a TF monitoring protocol based on a microscopy tool is justified, with benefits including rational use of medication, potential to generate savings, and reduced morbidity and mortality rates, in addition to enabling other diagnoses.

Effect of low radiation dose on the expression and location of aquaporins in rat submandibular gland.

Head and neck cancers are common in several regions of the world and the treatment usually includes radiotherapy. This treatment can generate adverse effects to the salivary flow, with a relationship between the dose and the damage caused. Salivary gland cells are highly permeable to water and therefore, they express aquaporins (AQPs). This study analyzed changes in the expression and location of these proteins and identified morphological changes induced by low radiation in rat submandibular gland. Female rats were divided into control and irradiated groups. Immunohistochemistry analysis allowed confirming the presence of AQP1 in the blood vessel endothelium. Intense and steady labelling granules were also observed in the cytoplasm of submandibular gland ductal cells. In addition, there was AQP5 positive labelling in ductal cells delimiting the lumen of intercalated duct, in the cytoplasm and membrane of acinar cells. Finally, the decrease of AQP labelling in irradiated animal glands validated their radiosensitivity. Thus, the decrease in AQP1 protein levels in the endothelium and AQP5 in gland ductal cells of irradiated animals may have hindered the removal of water from the lumen of ductal cells, inducing a delay in water absorption and triggering a slight lumen increase.

Avaliação da atividade antibacteriana, citotóxica e antioxidante da espécie vegetal Opuntia cochenillifera (L. ) Mill / Evaluation of antibacterial, cytotoxic, and antioxidant activity of the plant species Opuntia cochenillifera (L. ) Mill

RESUMO O presente estudo teve como objetivo avaliar a atividade antibacteriana, antioxidante e citotóxica da espécie Opuntia cochenillifera (L.) Mill. Foi realizada a prospecção fitoquímica e espectroscopia de absorção de infravermelho (IV) dos extratos etanólicos brutos e frações dos cladódios grande e pequeno. A atividade antioxidante foi avaliada pelo método da capacidade sequestradora de radicais livres utilizando o radical sintético 2,2-difenil-1-picrilhidrazila (DPPH). A atividade citotóxica foi obtida através do método colorimétrico do Metiltetrazolium (MTT). Já a atividade antibacteriana foi avaliada pelo método de microdiluição em caldo para determinar a concentração inibitória mínima (CIM) frente às estirpes bacterianas Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa e Escherichia coli. A prospecção fitoquímica revelou principalmente a presença de fenóis, esteroides livres, alcaloides, alcanos, além de outras classes químicas. O IV apresentou grupos funcionais como alcanos, carbonilas, grupos de metila, duplas ligações de carbono, grupamentos alquilamina, entre outros. Sobre a citotoxicidade na concentração de 100 μg/mL, os dois extratos brutos, todas as frações do cladódio grande e as frações de clorofórmio e metanol do cladódio pequeno não apresentaram toxicidade. Os extratos brutos e frações do cladódio grande e pequeno, não demonstraram atividade antibacteriana e nem antioxidante. Esses resultados podem fornecer suporte para pesquisas futuras, visando outras atividades biológicas da presente espécie vegetal.

ABSTRACT The purpose of this study was to evaluate the antibacterial, antioxidant, and cytotoxic activity of Opuntia cochenillifera (L.) Mill. A phytochemical screening and infrared (IR) absorption spectroscopy were performed in the crude ethanolic extracts and fractions of large and small cladodes. The antioxidant activity was evaluated through the qualitative method of free-radical scavenging capacity using the synthetic radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). The cytotoxic activity was obtained by the cell viability assay using methyl thiazolyl tetrazolium (MTT). Antibacterial activity was evaluated by broth microdilution method to determine the minimum inhibitory concentration (MIC) against the bacterial strains Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Escherichia coli. The phytochemical screening mainly revealed the presence of phenols, flavonoids, free steroids, alkaloids, alkanes, and other chemical classes. The IR spectroscopy presented functional groups such as alkanes, carbonyls, methyl groups, carbon double bonds, and alkylamino groups, among others. Regarding cytotoxicity in the concentration of 100 μg/mL, neither the crude extracts, the fractions of the large cladode, nor the chloroform and methanol fractions of small cladode presented toxicity. The crude ethanolic extracts and fractions of large and small cladode showed no antibacterial or antioxidant activity. These results may provide support for future research aimed at other biological activities of this plant species.

Low-temperature particulate calcium phosphates for bone regeneration.

BACKGROUND: Low-temperature synthesized calcium phosphates are produced by mixing calcium phosphate powders in an aqueous solution resulting in a precipitated phase. These compounds can be formulated in several forms (e.g. injectable cements and implantable blocks), and are commonly used as bone substitutes and drug delivery systems for the treatment of bone defects. As bone substitutes, calcium phosphates in general offer the advantages of being biocompatible and osteoconductive. AIMS: The present work employed a machine-based process to derive a reproducible preparation method for low-temperature calcium phosphate particulate (LTCP). The in vivo outcomes of LTCP were compared with those of three commercially available bone substitutes by histomorphometric measurements of bone formation and material degradation in a rat femur implantation model. MATERIALS & METHODS: Specifically, LTCP, anorganic bovine bone (AB), bioactive glass (BG), and demineralized bone matrix (DBM) were implanted in defects created in the distal aspect of rat femora. Reparative bone and particulate volumes of these biomaterials were evaluated post-operatively using micro-computed tomography and histological analyses at 3, 6, 12, and 16 weeks. RESULTS & DISCUSSION: Results showed that, despite invoking bone formation, AB, BG, and DBM were found un-resorbed in situ at 16 weeks. Conversely, LTCP showed an early increase in bone formation as well as clear evidence of complete degradation and reparative bone remodelling, resulting in the total reconstitution of the marrow cavity and marrow tissue. CONCLUSION: LTCP promoted increased early bone formation, associated with an improved degradation rate, compared with the other three bone-substitute biomaterials tested.

Prevalence of type 2 diabetes mellitus in Brazilian liver transplant candidates: negative association with HCV status.

Recently, an epidemiological association between hepatitis C virus (HCV) infection and type 2 diabetes mellitus (DM) has been reported in several studies, although many of them did not consider known risk factors in the pathogenesis of type 2 DM. The aim of this study was to assess the prevalence of type 2 DM among Brazilian HCV (+) and HCV (-) liver transplant candidates, analyzing known confounding factors for the development of type 2 DM. We conducted a cross-sectional study to evaluate the prevalence of type 2 DM among 106 liver transplant adult candidates, comparing 36 HCV (+) cirrhotic patients with 70 HCV (-) patients who developed cirrhosis from other causes. Type 2 DM was diagnosed after two consecutive fasting glucose values > or =126 mg/dL. The age, sex, and race distribution, severity of liver disease (Child-Pugh score), and family history of DM were similar in both groups, but the mean body mass index (BMI) was higher in the HCV (-) subjects (26.81 +/- 5.29 vs 24.0 +/- 4.71, P < .01) Most of the patients were Caucasians (70.75%). Type 2 DM was detected in 36.11% of HCV (+) group and in 25.71% of the HCV (-) (P = .27). A multivariate analysis revealed that family history of DM was the only significant independent predictor for DM (odds ratio = 2.55, 95% CI = 1.03 to 6.31, P = .04). In conclusion, our study did not show an association between HCV infection and Type 2 DM in Brazilian liver transplant candidates. It confirmed that the family history of DM was a determinant factor for the development of type 2 DM.

Prevalence of new-onset diabetes mellitus in Brazilian liver transplant recipients: association with HCV infection.

New-onset diabetes melittus (NODM) is a serious complication following transplantation. Recent studies suggest an association between hepatitis C virus (HCV) infection and DM both in nontransplant settings as well as after liver transplantation (LT). The aim of this study was to assess the prevalence of NODM among Brazilian LT recipients, analyzing possible risk factors including HCV infection. We conducted a cross-sectional study to evaluate the prevalence of NODM in 82 LT recipients with a posttransplant follow-up > or =1 year including 29 HCV-positive patients and 53 with other causes for liver disease. Patients were considered to meet the criteria for DM if they had two consecutive fasting glucose values > or =126 mg/dL or if they were taking insulin or oral hypoglycemic agents at the time of the study. The overall prevalence of NODM was 18.29% with a median interval of 20 months between LT and diagnosis of DM. The age, sex, and race distribution, immunosuppressive regimen, number of rejection episodes treated with pulse therapy, and family history of DM were similar in both groups. However, the frequency of BMI > or = 30 in the pre- and posttransplant periods was higher among patients who developed NODM (P = .02). Upon multivariate analysis of the entire cohort, HCV infection was the only significant predictor of NODM (OR = 4.31, CI = 1.17 to 15.84, P = .02). In conclusion, our study confirmed an association between HCV infection and NODM among Brazilian liver transplant recipients, suggesting that HCV infection may have a potential role in the pathogenesis of posttransplantation DM.

Determination of minocycline in human plasma by high-performance liquid chromatography coupled to tandem mass spectrometry: application to bioequivalence study.

Minocycline was determined in human plasma by HPLC-MS-MS using clarithromycin as an internal standard. The method is fast (single liquid extraction and run time of <3 min) and sensitive (5 ng/ml) and it was employed in a bioequivalence study of two 100 mg tablet formulations in 24 healthy volunteers. The 90% confidence interval of the individual ratio geometric mean for both AUC(0-96 h) and Cmax were 99.2-111.1% and 95.6-117.5%, respectively. Thus, Minoderm was considered bioequivalent to Minomax according to both the rate and extent of absorption. No food interaction was observed with either formulation.

[Number and thickness of myelinic fibres of the phrenic nerve of young and aged rats]. / Número e espessura de fibras mielínicas do nervo frênico de rato albino jovem e senil.

The phrenic nerve of albino rats was studied for age changes in number of fibres, myelin sheath thickness and axon calibre. There is no significant morphological differences between nerves from young and aged rats and no difference with age was found in the number of fibres, myelin sheath thickness and axon calibre.

[Visceral leishmaniasis epidemic in the State of Piauí, Brazil, 1980-1986]. / Epidemia de leishmaniose visceral no estado do Piauí, Brasil, 1980-1986.

The kala-azar epidemic in the State of Piauí 1980-1986 is analyzed on the basis of the data collected by SUCAM Piauí. The outbreak began in towns of central and northern Piauí in 1980. In contrast what has happened in endemic periods in which the disease occurred in areas of higher altitude and semi-arid climate, the epidemic developed in humid tropical river valleys in rural zones. The epidemic was worst in the towns. The state capital, Teresina, hit in 1981, reached the epidemic peak in 1984 and accounted, for more than 60% of the 1,509 cases in the state. The epidemic was not substantial in those regions sprayed to combat malaria and Chagas' disease. While control in Teresina was attempted through intensive use of insecticides, the outbreak gave way spontaneously in rural areas. Neither the number of cases nor the phlebotomine population of Teresina presented significant seasonal variations but were moderately correlated. There was greater prevalence in children of 5 years of age or less, especially during the peak epidemic years, and much lesser prevalence in adults over 40 years of age. The geographical distribution of the epidemic process and its beginning, concomitant with a prolonged drought with its accompanying migration of people and domestic animals from endemic to epidemic regions, suggests that migration unleashed the epidemic. The fact that the epidemic process spontaneously relinquished its hold in areas where no control was attempted, indicates that the end of the epidemic cannot be attributed solely to measures of control. An analysis of the coefficients of specific incidence within age groups sparks the discussion about the possibility that progressive reduction of susceptibility (determined by the great number of asymptomatic infections as well as by long-lasting immunity) contributed to the extinction of the epidemic.

Morphometric study of fat cell size in the fascia areolaris and fascia lamellaris of the inguinal region in men, women and pregnant women.

The fat cells of the fascia areolaris and fascia lamellaris of men, women, and pregnant women (aged between 20 and 35a) were morphometrically studied. The cell volumes showed the following average values: 4.423 X 10(5) micron3 and 2.004 X 10(5) micron3 for the fasciae areolaris and lamellaris respectively, in men; 6.236 X 10(5) micron3 and 3.964 X 10(5) micron3 in women, and 10.114 X 10(5) micron3 and 4.635 X 10(5) micron3 in the pregnant women. The analysis of variance showed significant differences between both sexes, and fasciae areolaris and lamellaris. The differences between women and pregnant women as far as the cell volume is concerned, in both fasciae, were not significant. As to the fascia areolaris, not the lamellaris, the difference between the sexes was significant.

[Biomphalaria glabrata in the state of Piaui]. / Biomphalaria glabrata no Estado do Piaui.

The occurrence of Biomphalaria glabrata is recorded for the first time in the state of Piaui, where it was collected from several breeding places in the city of Parnaíba. Examination of 694 specimens showed that a part of them were infected with trematodes other than Schistosomatidae. So far no autochthonous cases of schistosomiasis have been identified in the city. The presence of B. glabrata in Parnaíba extends by 20 km eastward its range on the Northern Coastal Region of the Great Northeastern Region of Brazil, where it had been found as far as Araioses, on the eastern extreme of the state of Maranhão.

Ultrastructural and morphometric study on fat cells of the so called subcutaneous "fascia areolaris" and "fascia lamelaris" in the human inguinal region.

The fat cells of the so called fascia areolaris and fascia lamelaris (Velpeau 1834; Sterzi 1910) of men and women (aged from 20 to 35 years) were ultrastructural and morphometrically (cell volume) studied. No noteworthy submicroscopic difference was observed between fascias. The cell volumes obtained from planimetric measures showed the following values: 3.770 X 10(5) microns and 2.497 X 10(5) microns in the fascia aerolaris and lamelaris of men, respectively. For the women the values were: 7.222 X 10(5) microns and 5.025 X 10(5) microns (Fig. 3). The analysis of variance shows significant differences between the sexes and between fascia areolaris and lamelaris. The difference between the fascias supports the Sterzi's (1910) description on the tela subcutanea as being formed by those two distinct layers.

Morphometric study of pre- and post-hatching nerve cell bodies of lumbar spinal ganglia of Gallus domesticus.

A cytomorphometric study was performed in lumbar spinal ganglia neurons of Gallus domesticus on the 10th and 18th incubation days and 8th, 35th, 61st, and 120th post-hatching days. The absolute volume of nucleus and relative volume of cytoplasm were respectively estimated by the Bach caryometric method and by point-counting volumetry, carried out in 0.5 micrometer thick araldite sections. The relative volume, the surface-to-volume ratio and the total surface of RER, SER, mitochondria, dense bodies, Golgi complex and the relative volume of hyaloplasm inside and outside the Nissl bodies were estimated from electronmicrographs by the Weibel et al. method. The conclusions were: a) there was an increase of the cell volume and a decrease of the nucleo-cytoplasmic ratio, particularly between the first two ages; b) the relative volumes of RER and SER change inversely with respect to each other: the RER increases before hatching, decreasing progressively afterwards; the changes of relative volume of dense bodies are similar to those of the RER, and the mitochondria show relatively small variations concerning the same parameter; c) the relative volume of hyaloplasm inside the Nissl bodies decreases while those outside increases; d) the surface-to-volume ratio drops sharply for all organelles from the 10th to the 18th day of incubation; after hatching, a tendency to increase is observed; e) the membrane surface-to-cytoplasmic volume ratio decreases for all organelles from the 10th to the 18th day of incubation; after hatching, this ratio increases slightly for mitochondria and Golgi complex, sharply for SER, dropping for dense bodies. The RER values alternate regularly.