Prevalence of oral and maxillofacial lesions in children and adolescents at a regional Brazilian oral pathology service: a retrospective study and the relevant literature review.

PURPOSE: This study assessed the prevalence of maxillofacial lesions in children, i.e., 0-9 years, and adolescents, i.e., 10-19 years, in a Brazilian Oral Pathology Service and compared results with available literature. METHODS: Clinical and histopathological records from January 2007 to August 2020 were analysed and a literature review investigating maxillofacial lesions in paediatric populations was also performed. RESULTS: Overall, "reactive salivary gland lesions" and "reactive connective tissue lesions" were the most prevalent group of soft tissue lesions, affecting children and adolescents equally. From these, mucocele and pyogenic granuloma were the most prevalent histological diagnoses, respectively, regardless of age. These findings were consistent with the 32 studies included. Considering intraosseous lesions, "odontogenic cysts" and "periapical inflammatory lesions" were the most prevalent groups, with no relevant differences between age groups, except for the odontogenic keratocyst, which was more prevalent in adolescents. Moreover, several odontogenic tumours, such as ameloblastic fibroma and odontogenic myxoma, were significantly more prevalent in children. CONCLUSION: Most maxillofacial lesions presented a similar prevalence between children and adolescents. Reactive salivary gland lesions and reactive connective tissue lesions were the prevailing diagnostic categories, regardless of age. Some odontogenic tumours and the odontogenic keratocyst showed significantly different frequencies across these age groups.

Shifts in uterine bacterial communities associated with endogenous progesterone and 17ß-estradiol concentrations in beef cattle.

The relation between circulating concentrations of progesterone and 17ß-estradiol prior to insemination play a key role in optimizing fertility in cattle. This study aimed to determine the impact of endogenous progesterone (P4) and estradiol (E2) concentrations on uterine bacterial community abundance and diversity in beef cattle. Angus-influenced heifers were subjected to an industry standard estrous synchronization protocol. Uterine flushes were collected on d -2 (endogenous P4) and d 0 (endogenous E2) and used for targeting the V4 hypervariable region of 16S rRNA bacterial gene. Plasma was collected on d -2 and 0 for quantification of P4 and E2 concentrations by radioimmunoassay, respectively. Heifers were allotted to one of the following groups: High P4 + High E2 (H-H; n = 11), High P4 + Low E2 (H-L; n = 9), Low P4 + High E2 (L-H; n = 9), Low P4 + Low E2 (L-L; n = 11). Results indicated that Shannon's diversity index tended to be greater for H-L heifers compared to L-H heifers on d 0 (P = 0.10). For H-L heifers from d -2 to d 0, the relative abundance of Actinobacteria decreased and Tenericutes increased (P < 0.01). Within phylum Actinobacteria, the relative abundance of Corynebacterium decreased from d -2 to d 0 in treatment groups H-H, H-L, and L-L (P < 0.05); however, did not differ by d for L-H heifers. Within phylum Tenericutes, the relative abundance of Ureaplasma increased from d -2 to d 0 for H-L heifers (P = 0.01). Additionally for H-L heifers, the relative abundance of Bacteroidetes tended to increase from day -2 to on d 0 (P = 0.07). For H-L heifers, uterine pH increased from day -2 to d 0 (P = 0.05). These results suggest that differing endogenous concentrations of P4 and E2 may be associated with shifts in uterine microbiota and pH, and this could ultimately impact fertility outcomes in beef cattle.

Pregnancy maintenance following sequential induced prostaglandin pulses in beef cows.

The relationship between the maternal endocrine environment and late embryonic mortality (> 28 d of gestation) in cattle is poorly defined. A definitive rise and alterations in secretion patterns of prostaglandin F2α (PGF2α) concentration without luteal regression is a trademark of this period. The objective was to evaluate whether consecutively induced PGF2α pulses would alter steroid hormone production and luteal blood perfusion potentially influencing pregnancy success. Pregnant beef cows (n = 12) were selected to receive either an oxytocin injection (OT, n = 8) or saline injection (CON, n = 4) on d 30 and 31 of gestation to stimulate sequential prostaglandin releases 24 h apart. Blood samples were collected every 30 min for 1 h before and continuing for 4 h post oxytocin administration. Luteal blood perfusion was measured via Doppler ultrasound at the beginning and end of the OT challenge. Concentrations of prostaglandin F2α metabolite (PGFM) were quantified to show effectiveness of the treatment while concentrations of progesterone, estradiol and pregnancy-associated glycoproteins (PAG) were measured to examine the effect of PGF2α release. Control animals exhibited no changes in any quantified hormone and an expected numerical increase in circulating PAG concentrations. Peak concentrations of PGFM in OT cows were observed 2 h post OT administration and concentrations returned to basal levels by the end of the sampling period. Peak concentrations of PGFM were decreased on d 31 compared to d 30. Following OT administration, progesterone and estradiol concentrations did not change in response to PGF2α release but were decreased on d 31 compared to d 30. There were no changes in luteal blood perfusion in response to PGF release on d 30 or d 31. Repeated PGF2α release may alter steroid hormone production; however, it does not negatively affect pregnancy status during the transition between early and late embryonic development.

Effectiveness of marsupialisation and decompression on the reduction of cystic jaw lesions: a systematic review.

In this study, we aimed to systematically review and critically appraise the available literature concerning the effectiveness of marsupialisation and decompression on the reduction of cystic jaw lesions. The 'Preferred Reporting Items for Systematic Reviews and Meta-Analysis' guidelines were followed and the study protocol was registered at the 'International Prospective Register of Systematic Reviews' (CRD42019116099). Six main databases were searched: Embase, LILACS, PubMed, Scopus, The Cochrane Library, and Web of Science. Searches were complemented with three grey literature sources: Google Scholar, ProQuest, and Open Grey. Any reduction measures, compared with preoperative status or other procedures, were considered. Risk of bias was assessed using the Joanna Briggs Institute Critical Appraisal Tool. Thirty-one studies were included, of which five were judged with low, 24 with moderate, and two with high risk of bias. Considering surgical approach, 20 studies assessed the decompression and 11 the marsupialisation technique. Most studies considered these techniques as preliminary treatments, followed by enucleation. From 1088 lesions found, most were odontogenic keratocysts (33.8%), followed by unicystic ameloblastomas (21.0%), dentigerous cysts (20.6%), and radicular cysts (8.4%). Large lesions and younger individuals frequently presented more favourable responses to treatment and anatomical location was not associated with lesion reduction overall. The intervention duration generally ranged between one to two years. In conclusion, marsupialisation and decompression were mostly considered as preliminary treatments, followed by enucleation. Lesion reduction was generally considered insufficient for these techniques to be used as definitive therapies, although benefits concerning the diminished invasiveness of the secondary surgery were often proposed.

Effects of estradiol treatments on PGF2α release in beef heifers submitted to estrous resynchronization 14 days after timed-AI.

The effects of 17ß-estradiol (E2) or estradiol benzoate (EB) on PGF2α release were studied in bred-non-pregnant and pregnant Nelore beef heifers. The day of timed artificial insemination (TAI) was designated day 0 (D0), and a single treatment was given on D14. All heifers also received an intravaginal P4 device on D14, and were randomly assigned to three groups: Control (C, P4 device only, n = 12); E2 (1 mg E2 + 9 mg P4, n = 10); or EB (1 mg, n = 10). Blood samples were collected hourly for 8 hours after treatment (Hours 0-8) to measure plasma concentrations (pg/mL) of a PGF2α metabolite (PGFM). The P4 device was removed on D22 and pregnancy was diagnosed on D28. Pregnancy rate was not different among groups (C, n = 7/12; E2, n = 5/10; EB, n = 5/10). More (P < 0.05) heifers had a CV-identified prominent PGFM pulse (peak of > 100 pg/mL) in E2 group (6/10) than in EB (1/10) and C (0/12) groups. Hourly concentration of PGFM for Hours 0 to 8 showed significant effects of group and hour and an interaction of group by hour but did not show an interaction of group or hour with pregnancy status. In preliminary post-hoc analyses, PGFM concentrations during Hours 0 to 8 and pulse characteristics were analyzed within each pregnancy status. For the non-pregnant heifers, a group-by-hour interaction was detected tentatively indicating an increase (P < 0.005) in PGFM concentrations in E2 group from Hours 4 to 6 and in EB group at Hours 5 and 6. Maximum PGFM concentration during Hours 0 to 8 did not differ (P > 0.1) between E2 (124 ± 23) and EB (110 ± 30) groups, but was greater (P < 0.05) in each group than in C (32 ± 3). Furthermore, PGFM concentrations of pulses at the peak, amplitude, and area under pulse curve (pg/mL/h) were greater (P < 0.05) in E2 group than in C group whereas the EB group did not differ (P > 0.1) from the other groups. For pregnant heifers, no effects of group, hour, or their interaction were detected in PGFM concentrations during the hourly sessions, except that maximum PGFM concentration was greater (P < 0.05) in E2 than in EB and C groups. In addition, the number of prominent pulses was greater in E2 group than in Control or EB groups. In conclusion, PGFM increased earlier and in greater concentration combined for bred-non-pregnant and pregnant heifers treated 14 days after TAI with 1 mg E2 plus 9 mg P4 than with 1 mg EB. Tentatively, a positive effect for each of E2 and EB on PGFM concentrations was attenuated in pregnant heifers.

Type I interferon receptors and interferon-τ-stimulated genes in peripheral blood mononuclear cells and polymorphonuclear leucocytes during early pregnancy in beef heifers.

This study characterised the expression of interferon (IFN)-τ-stimulated genes (ISGs) and Type I IFN receptors in circulating polymorphonuclear cells (PMNs) of beef heifers and compared it with expression in peripheral blood mononuclear cells (PBMCs) up to Day 20 of gestation. Nelore heifers (n=26) were subjected to fixed-time AI (FTAI) on Day 0. PMNs and PBMCs were isolated on Days 0, 10, 14, 16, 18 and 20 after FTAI. The abundance of target transcripts (ubiquitin-like protein (ISG15), 2'-5'-oligoadenylate synthetase 1 (OAS1), myxovirus resistance 1 (MX1), myxovirus resistance 2 (MX2), IFN receptor I (IFNAR1) and IFN receptor 2 (IFNAR2)) was determined using real-time quantitative polymerase chain reaction and compared between pregnant (n=8) and non-pregnant (n=9) females. In both PBMCs and PMNs, ISG15 and OAS1 expression was greater in pregnant than non-pregnant heifers on Days 18 and 20. There were no significant differences in the expression of ISGs between PBMCs and PMNs. A time effect on expression was found for IFNAR1 in PBMCs and IFNAR2 in PMNs, with decreased expression of both genes on Days 18 and 20. When the expression of these genes was compared between cell types only in pregnant heifers, IFNAR2 expression in PMNs had an earlier decrease when compared to its expression in PBMCs, starting from Day 18. In conclusion, PMNs do not respond earlier to the conceptus stimulus, and ISG15 and OAS1 expression in both PMNs and PBMCs can be used as a suitable marker for pregnancy diagnosis on Days 18 and 20. In addition, gestational status did not affect IFNAR1 and IFNAR2 expression, but IFNAR2 showed a distinct response between PMNs and PBMCs of pregnant heifers.

Increased pregnancy rate in beef heifers resynchronized with estradiol at 14 days after TAI.

We aimed to evaluate the treatment with estradiol benzoate (EB) or 17ß-estradiol (E2) associated with progesterone (P4) for resynchronization of ovulation 14 days after timed artificial insemination (TAI). In Experiment 1 (Exp. 1), Nelore heifers were submitted to TAI (D0). On D14, the animals received an intravaginal P4 device and were randomly assigned to one of three groups: control (no treatment; n = 17); EB (1  mg EB; n = 17); and E2+P4 (1 mg E2 + 9 mg P4; n = 18). Ultrasonography evaluations were performed daily from D14 to D22 to map follicular and luteal dynamics. On D22, the P4 devices were removed and non-pregnant (NP) animals were determined using corpus luteum blood flow Doppler ultrasonography. In Exp. 2, 1295 beef heifers were resynchronized and randomly allocated to the same experimental groups as described in Exp. 1. On D22, the largest follicle (LF) was measured in NP and a second TAI was performed on D24. In a subset of heifers (n = 337), an estrus detection patch was used between D22 and D24 to monitor estrus expression and the LF was measured at D24. Confirmatory diagnosis of pregnancy was performed between D37-67 and D43-67 after first and second TAI, respectively. In Exp 1, the proportion of heifers with a synchronized follicular wave emergence (from 3 to 5 days after treatment) was greater (P < 0.05) in the EB group (93.8%) than in the control (62.5%) and E2+P4 (64.7%) groups. Structural luteolysis occurred earlier (P < 0.05) in the EB and E2+P4 groups than in the controls. The pregnancy rate after first TAI did not differ (P > 0.1) among the groups at D22 and at confirmatory diagnosis in both experiments. In Exp 2, the potential pregnancy loss between D22 and D37-67 was similar (P > 0.1) in the control (19% [36/185]), EB (15% [28/182]) and E2+P4 (15% [28/184]) groups. The LF diameter (mm) on D22 was greater (P < 0.05) in the control group (11.9 ± 0.1) than in EB (11.3 ± 0.1) and E2+P4 (11.5 ± 0.1). No difference (P > 0.1) was observed in the proportion of heifers detected in estrus, but LF growth rate (mm/day) between D22 and D24 was greater (P < 0.05) in EB group (0.9 ± 0.08) than in control (0.6 ± 0.07) and E2+P4 (0.7 ± 0.09) groups. The pregnancy rate for the second TAI was greater (P < 0.05) in the EB group (47% [94/200]) than in the control (37% [76/203]), but did not differ (P > 0.1) from the E2+P4 group (43% [93/214]). In conclusion, the treatment with 1 mg EB or 1 mg E2 + 9 mg P4 at 14 days post-TAI anticipates luteolysis in NP heifers but does not compromise pregnancy. The EB treatment induces a new synchronized follicle wave emergence and increases the pregnancy rate of resynchronized NP heifers.

Metaloproteinases no tecido laminar do casco de equinos submetidos à obstrução intraluminal do cólon menor / Metalloproteinases in hoof laminar tissue of equine submitted to intraluminal small colon obstruction

O objetivo deste estudo foi avaliar a expressão das MMP-2 e MMP-9 no tecido laminar do casco e o perfil leucocitário de equinos submetidos à obstrução intraluminal do cólon menor. Realizaram-se laparotomia e obstrução do cólon menor de oito equinos hígidos, utilizando-se uma bola inserida no lúmem intestinal. A bola foi inflada à pressão de 80mmHg e a obstrução foi mantida por quatro horas. Foram realizadas coletas sanguíneas antes da obstrução (M0), imediatamente após a desobstrução (M4) e a cada 12 horas após M4, até completar 72 horas (M12, M24, M36, M48, M60 e M72). As biópsias de casco foram realizadas em M0, M4 e M72, e as amostras foram submetidas à análise zimográfica. Foi observado aumento nos leucócitos em M12 e M24, decorrente do aumento de neutrófilos segmentados e bastonetes, os quais diminuíram a partir de M36. Segundo a técnica zimográfica, não se observaram alterações nos valores de MMP-2 e -9, possivelmente devido à baixa intensidade das lesões ocasionadas no cólon menor. Com isso, conclui-se que as alterações inflamatórias decorrentes da obstrução do cólon menor não foram suficientes para ocasionar alterações na expressão das MMP-2 e -9 no tecido laminar podal.(AU)

The aim of this study was to evaluate the blood leukocytes and the MMP-2 and -9 expression in the hoof laminar tissue of horses undergoing intraluminal small colon obstruction. Laparotomy and the small colon obstruction was performed in eight healthy horses, inserting a ball in the intestinal lumen. The ball was inflated to 80 mmHg pressure and the occlusion was maintained for 4 hours. The blood was collectedBlood samples were taken before the obstruction (M0), immediately after intestinal clearance (M4), and every 12 hours until completeuntil 72 hours (M12, M24, M36, M48, M60 and M72). The hoof biopsies were performed at M0, M4, and M72 and the samples were subjected to zymography analysis. There was an increase in leukocytes in M12 and M24, due to the increase in segmented neutrophils and band neutrophils, which decreased as of M36. According to zymography technique not observed changes were not not observed in MMP-2 and -9, possibly due to the low intensity of the small colon lesions. Wherefore, it is concludedIn conclusion, that the inflammatory changes resulting from small colon obstruction were not enough to cause changes in the expression of MMP-2 and -9 in the hoof laminar tissue.(AU)

Variantes anatómicas del canal carpiano y distancias de estructuras vasculares en ecografía / Anatomical variants the carpal tunnel and distances of vascular structures in ultrasound

El síndrome de canal carpiano es una patología frecuente. Si bien el diagnóstico es clínico, la ecografía cumple un rol en caso de duda diagnóstica y como apoyo a proce-dimientos intervencionales.Existen variables anatómicas y distancias de estructuras vasculares útiles de conocer antes de planear un gesto quirúrgico o de infiltración para disminuir el riesgo de lesiones secundarias, en donde la ecografía podría tener un rol.Estudiamos una muestra de 267 ecografías de muñeca con especial hincapié en va-riantes neurogénicas, vasculares o tendinosas que podrían resultar lesionadas en relación a algún procedimiento.

Carpal tunnel syndrome is a frequent pathology. Although the diagnosis is clinical, ultrasound plays a role in case of diagnostic doubt and as support and guide for inter-ventional procedures.There are anatomical variants and distances of vascular structures that may be useful to know before planning a surgical or infiltration procedure to reduce the risk of iat-rogenic injuries, where ultrasound could play a role.We studied a sample of 267 wrists ultrasounds with special emphasis on neurogenic, vascular or tendinous variants that could be injured in relation to procedures.

Inhibitory effect of the Pseudobrickellia brasiliensis (Spreng) R.M. King & H. Rob. aqueous extract on human lymphocyte proliferation and IFN-γ and TNF-α production in vitro.

Pseudobrickellia brasiliensis (Asteraceae) is a plant commonly known as arnica-do-campo and belongs to the native flora of the Brazilian Cerrado. The alcoholic extract of the plant has been used as an anti-inflammatory agent in folk medicine, but the biological mechanism of action has not been elucidated. The present study evaluated the composition of P. brasiliensis aqueous extract and its effects on pro-inflammatory cytokine production and lymphocyte proliferation. The extracts were prepared by sequential maceration of P. brasiliensis leaves in ethanol, ethyl acetate, and water. Extract cytotoxicity was evaluated by trypan blue exclusion assay, and apoptosis and necrosis were measured by staining with annexin V-FITC and propidium iodide. The ethanolic (ETA) and acetate (ACE) extracts showed cytotoxic effects. The aqueous extract (AQU) was not cytotoxic. Peripheral blood mononuclear cells stimulated with phorbol myristate acetate and ionomycin and treated with AQU (100 µg/mL) showed reduced interferon (IFN)-γ and tumor necrosis factor (TNF)-α expression. AQU also inhibited lymphocyte proliferative response after nonspecific stimulation with phytohemagglutinin. The aqueous extract was analyzed by liquid chromatography coupled with photodiode array detection and mass spectrometry. Quinic acid and its derivatives 5-caffeoylquinic acid and 3,5-dicaffeoylquinic acid, as well as the flavonoids luteolin and luteolin dihexoside, were detected. All these compounds are known to exhibit anti-inflammatory activity. Taken together, these findings demonstrate that P. brasiliensis aqueous extract can inhibit the pro-inflammatory cytokine production and proliferative response of lymphocytes. These effects may be related to the presence of chemical substances with anti-inflammatory actions previously reported in scientific literature.

Exigências de energia de borregas mestiças alimentadas com níveis crescentes de concentrado na dieta / Energy requirements of crossbreed ewe lambs fed with increasing dietary levels of concentrate

Objetivou-se estimar as exigências energéticas de borregas mestiças confinadas, alimentadas com níveis crescentes de concentrado na dieta. Foram avaliadas 36 borregas, com peso corporal inicial de 23,7±3,67kg. Seis animais foram abatidos no início do experimento para obtenção dos valores do grupo referência. Os demais (30 animais) foram distribuídos em cinco tratamentos: mantença (alimentação restrita com feno de capim Tifton) e suplementados com níveis crescentes de concentrado (20, 40, 60 e 80%) em base de matéria seca (seis animais por tratamento). As borregas foram abatidas quando atingiram 37,70±9,89kg. Os níveis de concentrado influenciaram a eficiência de utilização da energia metabolizável para mantença (km) e consequentemente as exigências diárias de energia metabolizável para mantença. A exigência de energia líquida para mantença de borregas mestiças em crescimento é 57 kcal/PCJ0,75/dia, em que PCJ é o peso corporal em jejum. As borregas com 20kg apresentaram exigência de energia líquida para 100g de ganho de peso diário de 465kcal/dia. As borregas com 40kg de peso corporal exigiram 930kcal/dia de energia líquida para o mesmo ganho. A exigência líquida para mantença de borregas pode ser estimada pela equação: ELm (Mcal/dia) = 0,057* PCJ0,75. A exigência líquida de energia para ganho de borregas mestiças (Mcal/dia) pode ser estimada a partir da equação: ELg = 0,524 x PVJ0,75 x GPCVZ1,21, em que GPCVZ é o ganho de peso do corpo vazio. As dietas influenciam as eficiências de usos da energia metabolizável para mantença (km) e ganho (kg).(AU)

The aim of this study was to estimate the energy requirements of crossbreed ewe lambs in a feedlot fed with increasing levels of concentrate in the diet. 36 ewe lambs were evaluated with initial body weight of 23.7±3.67kg. Six animals were slaughtered at the beginning of the experiment to obtain the reference group values. Animals (30) were distributed in five treatments: maintenance (feed restricted with Tifton grass hay) and those supplemented with increasing dietary levels of concentrate (20, 40, 60 and 80%) on a dry matter basis (six animals per treatment). The ewe lambs were slaughtered when they reached 37.70±9.89kg. The concentrate levels influenced the utilization efficiency of metabolizable energy for maintenance (km) and consequently the daily requirements of metabolizable energy for maintenance. The net energy requirements for maintenance of crossbred lambs in growth is 57kcal/FCW 0.75/day, where FCW is body weight on fasting. The ewe lambs with 20kg presented net energy requirement for 100g daily weight gain of 465kcal/day. The ewe lambs with 40kg of body weight required 930 kcal/day of net energy for the same gain. The net energy requirement for maintenance of ewe lambs can be estimated by the equation: NEm (Mcal/day) = 0.057 * FCW0.75. The net energy requirement for crossbred ewe lambs gain (Mcal/day) can be estimated from the equation: NEg (Mcal/day) = 0.524*FCW 0.75*GEBW1.21, where GEBW represented gain empty body weight. Diets affect the efficiencies of uses of metabolizable energy for maintenance (km) and gain (kg).(AU)

Amniotic membrane as a biological dressing for 5-fluoruracil-induced oral mucositis in rats.

Oral mucositis is a reaction to chemoradiation therapy during cancer treatment. The aim of this study was to investigate the use of amniotic membrane as a biological dressing for oral mucositis lesions in rats. Sixty Wistar rats were divided into three groups (n = 20): control, 5-fluoruracil (5-FU), 5-fluoruracil + amniotic membrane (5-FU+AM). Each group was subdivided (n = 5) according to the time interval to sacrifice (3, 7, 14, and 21 days). Histology (haematoxylin-eosin staining) and immunocytochemistry (anti-rat antibodies CD4, CD8, VEGF, and PCNA) were evaluated. Immunocytochemistry results were analyzed using one-way ANOVA and Tukey tests. The amniotic membrane (5FU+AM) played an important role in cell proliferation (PCNA 3 days 27.08 ± 4.65, 7 days 27.90 ± 3.34) and especially in neovascularization (VEGF 3 days 23.00 ± 1.40, 7 days 26.00 ± 0.95) for all time intervals, when compared to 5-FU (PCNA 3 days 23.12 ± 1.61, 7 days 37.21 ± 1.20; VEGF 3 days 17.05 ± 1.51, 7 days 8.45 ± 1.35) and control (PCNA 3 days 29.99 ± 0.92, 7 days 16.33 ± 2.88; VEGF 3 days 13.65 ± 0.55, 7 days 15.70 ± 1.39). It was biocompatible, showing significant differences compared to the other groups in CD4 (F = 40.72; P = 0.001) and CD8 (F = 69.99, P = 0.001) staining together, only during the inflammation phase (7 days). Amniotic membrane presented biocompatibility and stimulated cell proliferation and neovascularization, functioning as a promising biological dressing.

Invariant Natural Killer T Cells are Reduced in Hereditary Hemochromatosis Patients.

PURPOSE: Invariant natural killer T (iNKT) cells are CD1d restricted-T cells that react to lipid antigens. iNKT cells were shown to be important in infection, autoimmunity and tumor surveillance. Alterations in the number and function of these cells were described in several pathological conditions including autoimmune and/or liver diseases. CD1d is critical for antigen presentation to iNKT cells, and its expression is increased in liver diseases. The liver is the major organ affected in Hereditary Hemochromatosis (HH), an autosomal recessive disorder caused by excessive iron absorption. Herein, we describe the study of iNKT cells of HH patients. METHODS: Twenty-eight HH patients and 24 control subjects from Santo António Hospital, Porto, were included in this study. Patient's iron biochemical parameters (serum transferrin saturation and ferritin levels) and the liver function marker alanine transaminase (ALT) were determined at the time of study. Peripheral blood iNKT cells were analyzed by flow cytometry using an anti-CD3 antibody and the CD1d tetramer loaded with PBS57. RESULTS: We found a decrease in the percentage and number of circulating iNKT cells from HH patients when compared with control population independently of age. iNKT cell defects were more pronounced in untreated patients, relating with serum ferritin and transferrin saturation levels. No correlation was found with ALT, a marker of active liver dysfunction. CONCLUSIONS: Altogether, our results demonstrate that HH patients have reduced numbers of iNKT cells and that these are influenced by iron overload.

Trypan blue exclusion assay by flow cytometry

Dye exclusion tests are used to determine the number of live and dead cells. These assays are based on the principle that intact plasma membranes in live cells exclude specific dyes, whereas dead cells do not. Although widely used, the trypan blue (TB) exclusion assay has limitations. The dye can be incorporated by live cells after a short exposure time, and personal reliability, related to the expertise of the analyst, can affect the results. We propose an alternative assay for evaluating cell viability that combines the TB exclusion test and the high sensitivity of the flow cytometry technique. Previous studies have demonstrated the ability of TB to emit fluorescence when complexed with proteins. According to our results, TB/bovine serum albumin and TB/cytoplasmic protein complexes emit fluorescence at 660 nm, which is detectable by flow cytometry using a 650-nm low-pass band filter. TB at 0.002% (w/v) was defined as the optimum concentration for distinguishing unstained living cells from fluorescent dead cells, and fluorescence emission was stable for 30 min after cell treatment. Although previous studies have shown that TB promotes green fluorescence quenching, TB at 0.002% did not interfere with green fluorescence in human live T-cells stained with anti-CD3/fluorescein isothiocyanate (FITC) monoclonal antibody. We observed a high correlation between the percentage of propidium iodide+CD3/FITC+ and TB+CD3/FITC+ cells, as well as similar double-stained cell profiles in flow cytometry dot-plot graphs. Taken together, the results indicate that a TB exclusion assay by flow cytometry can be employed as an alternative tool for quick and reliable cell viability analysis.

Evaluation of anti-nociceptive and anti-inflammatory activity of low-level laser therapy on temporomandibular joint inflammation in rodents.

The aim of this study was to investigate the analgesic and anti-inflammatory activity of low-level laser therapy (LLLT) on the nociceptive behavioral as well as histomorphological aspects induced by injection of formalin and carrageenan into the rat temporomandibular joint. The 2.5% formalin injection (FRG group) induced behavioral responses characterized by rubbing the orofacial region and flinching the head quickly, which were quantified for 45 min. The pretreatment with systemic administration of diclofenac sodium-DFN group (10 mg/kg i.p.) as well as the irradiation with LLLT infrared (LST group, 780 nm, 70 mW, 30 s, 2.1 J, 52.5 J/cm(2), GaAlAs) significantly reduced the formalin-induced nociceptive responses. The 1% carrageenan injection (CRG group) induced inflammatory responses over the time-course of the study (24 h, and 3 and 7 days) characterized by the presence of intense inflammatory infiltrate rich in neutrophils, scanty areas of liquefactive necrosis and intense interstitial edema, extensive hemorrhagic areas, and enlargement of the joint space on the region. The DFN and LST groups showed an intensity of inflammatory response that was significantly lower than in CRG group over the time-course of the study, especially in the LST group, which showed exuberant granulation tissue with intense vascularization, and deposition of newly formed collagen fibers (3 and 7 days). It was concluded that the LLLT presented an anti-nociceptive and anti-inflammatory response on the inflammation induced in the temporomandibular joint of rodents.

Substituição de monensina sódica por bicarbonato de sódio em dietas de novilhas confinadas / Evaluation of the replacement of Monensin with Sodium Bicarbonate in diets for confined bovines

Avaliaram-se os efeitos da substituição da monensina sódica pelo bicarbonato de sódio em dietas de novilhas em confinamento em relação ao desempenho animal, à conversão alimentar e ao parâmetro de fermentação ruminal. Foram utilizadas 56 novilhas mestiças, confinadas por 112 dias, submetidas a dietas com duas porcentagens de concentrado, 50% e 80% da matéria seca, com adição de monensina sódica ou bicarbonato de sódio, em fatorial 2x2. O volumoso utilizado para compor as dietas testadas foi composto de silagem de milho e cana-de-açúcar na proporção de 60:40, respectivamente. O nitrogênio amoniacal foi maior nos animais alimentos com dietas com alto grão. A monensina, na dieta 80:20, proporcionou maiores valores de nitrogênio amoniacal, uma e nove horas após o consumo da ração. O uso de bicarbonato promoveu os maiores valores de pH ruminal na dieta com 50% de concentrado, de seis a 18 horas após o consumo. Não houve efeito do aditivo sobre o desempenho animal. A alta inclusão de grãos na dieta propiciou aumento do desempenho animal e da eficiência alimentar. Conclui-se que o uso de bicarbonato de sódio em dietas para terminação de animais em confinamento é uma opção viável em substituição à monensina sódica para a manipulação da fermentação ruminal.

The effects of replacing monensin with sodium bicarbonate in diets for confined heifers on weight gain (WG), dry matter intake (DMI), feed:gain ratio (FGR) and rumen fermentation parameters were evaluated. Fifity six heifers were utilized (Santa Gertrudis x Braunvieh). Diets were formulated with two levels of concentrate (50% and 80% concentrate) with sodium monensin or sodium bicarbonate. The DMI was higest in heifers with an intake of 80% concentrate. There was not difference when the same concentrate levels were evaluated in relation to the DMI. The heifers at 80% concentrate showed highest WG. There was no significant difference in relation to WG between monensin and bicarbonate treatments and levels concentrate proportions. There were significant effects regarding FGR between treatments. The heifers on 80% concentrate showed lowest FGR. In the evaluation of N-NH3, the highest values were observed, on the whole, at 80:20 diets in relation to 50:50 diets. In the additive use, the monensin on 80:20 diets promoted the highest values of N-NH3 at 1 for 9 hours after feed intake. In the values of ruminal pH, the bicarbonate increased the values on 50:50 at 6 for18 hours after feed intake, in relation to the 80:20 diet. In this case, it was concluded that the use of sodium bicarbonate in diets for confined finishing heifers is a viable option for the replacement of monensin sodium.

Whole-body vibration decreases the proliferativeb response of TCD4+ cells in elderly individuals with knee osteoarthritis

The aim of this study was to investigate the effect of adding whole-body vibration (WBV; frequency = 35 to 40 Hz; amplitude = 4 mm) to squat training on the T-cell proliferative response of elderly patients with osteoarthritis (OA) of the knee. This study was a randomized controlled trial in which the selected variables were assessed before and after 12 weeks of training. Twenty-six subjects (72 ± 5 years of age) were divided into three groups: 1) squat training with WBV (WBV, N = 8); 2) squat training without WBV (N = 10), and 3) a control group (N = 8). Women who were ≥60 years of age and had been diagnosed with OA in at least one knee were eligible. The intervention consisted of 12 uninterrupted weeks of squatting exercise training performed 3 times/week. Peripheral blood mononuclear cells were obtained from peripheral blood collected before and after training. The proliferation of TCD4+ and TCD8+ cells was evaluated by flow cytometry measuring the carboxyfluorescein succinimidyl ester fluorescence decay before and after the intervention (∆). The proliferative response of TCD4+ cells (P = 0.02, effect size = 1.0) showed a significant decrease (23%) in the WBV group compared to the control group, while there was no difference between groups regarding the proliferative response of TCD8+ cells (P = 0.12, effect size = 2.23). The data suggest that the addition of WBV to squat exercise training might modulate T-cell-mediated immunity, minimizing or slowing disease progression in elderly patients with OA of the knee.

Expression of the chemokine receptor CXCR4 on lymphocytes of leprosy patients

Leprosy is caused by Mycobacterium leprae, which induces chronic granulomatous infection of the skin and peripheral nerves. The disease ranges from the tuberculoid to the lepromatous forms, depending on the cellular immune response of the host. Chemokines are thought to be involved in the immunopathogenesis of leprosy, but few studies have investigated the expression of chemokine receptors on leukocytes of leprosy patients. In the present study, we evaluated 21 leprosy patients (M/F: 16/5) with a new diagnosis from the Dermatology Outpatient Clinic of the University Hospital, Federal University of Minas Gerais. The control group was composed of 20 healthy members (M/F: 15/5) of the community recruited by means of announcements. The expression of CCR2, CCR3, CCR5, and CXCR4 was investigated by flow cytometry on the surface of peripheral blood lymphocytes. There was a decrease in percentage of CD3+CXCR4+ and CD4+CXCR4+ lymphocytes in the peripheral blood of leprosy patients (median [range], 17.6 [2.7-41.9] and 65.3 [3.9-91.9], respectively) compared to the control group (median [range], 43.0 [3.7-61.3] and 77.2 [43.6-93.5], respectively). The percentage of CD4+CXCR4+ was significantly lower in patients with the tuberculoid form (median [range], 45.7 [0.0-83.1]) of the disease, but not in lepromatous patients (median [range], 81.5 [44.9-91.9]). The CXCR4 chemokine receptor may play a role in leprosy immunopathogenesis, probably directing cell migration to tissue lesions in tuberculoid leprosy patients.

Expression of the chemokine receptor CXCR4 on lymphocytes of leprosy patients.

Leprosy is caused by Mycobacterium leprae, which induces chronic granulomatous infection of the skin and peripheral nerves. The disease ranges from the tuberculoid to the lepromatous forms, depending on the cellular immune response of the host. Chemokines are thought to be involved in the immunopathogenesis of leprosy, but few studies have investigated the expression of chemokine receptors on leukocytes of leprosy patients. In the present study, we evaluated 21 leprosy patients (M/F: 16/5) with a new diagnosis from the Dermatology Outpatient Clinic of the University Hospital, Federal University of Minas Gerais. The control group was composed of 20 healthy members (M/F: 15/5) of the community recruited by means of announcements. The expression of CCR2, CCR3, CCR5, and CXCR4 was investigated by flow cytometry on the surface of peripheral blood lymphocytes. There was a decrease in percentage of CD3+CXCR4+ and CD4+CXCR4+ lymphocytes in the peripheral blood of leprosy patients (median [range], 17.6 [2.7-41.9] and 65.3 [3.9-91.9], respectively) compared to the control group (median [range], 43.0 [3.7-61.3] and 77.2 [43.6-93.5], respectively). The percentage of CD4+CXCR4+ was significantly lower in patients with the tuberculoid form (median [range], 45.7 [0.0-83.1]) of the disease, but not in lepromatous patients (median [range], 81.5 [44.9-91.9]). The CXCR4 chemokine receptor may play a role in leprosy immunopathogenesis, probably directing cell migration to tissue lesions in tuberculoid leprosy patients.

Microbial profile and antibiotic susceptibility of culture-positive bacterial endophthalmitis.

PURPOSE: To assess the distribution of microorganisms isolated from patients with bacterial endophthalmitis and their antimicrobial susceptibility. METHODS: Retrospective analysis of medical and microbiological records of patients with suspected diagnosis of endophthalmitis. The following information was assessed: number of presumed and culture-positive endophthalmitis cases, source of infection, microbiological result (aqueous and/or vitreous culture and Gram staining), microbial characterization and distribution, and antimicrobial susceptibility. RESULTS: A total of 107 (46%) of 231 patients with bacterial endophthalmitis showed positive results by gram stain or culture. Of these, 97 (42%) patients were positive for culture only. Most of them (62%) were secondary to a surgical procedure (postoperative), 12% were posttraumatic and 26% were secondary to an unknown source or the data were unavailable. A total of 100 microorganisms were isolated (38 aqueous and 67 vitreous samples) from the 97 culture-positive cases (91% were gram-positive and 9% were gram-negative). Coagulase-negative Staphylococcus(CoNS) (48%) were the most frequently isolated, followed by Stretococcus viridans(18%), and Staphylococcus aureus(13%). The antimicrobial susceptibility for CoNS was as follows: amikacin-91.6%, cephalothin-97.9%, ceftriaxone-50%, ciprofloxacin-62.5%, chloramphenicol-91.8%, gatifloxacin-79.5%, gentamicin-72.9%, moxifloxacin-89.5%, ofloxacin-70.8%, oxacillin-58.3%, penicillin-33.3%, tobramycin-85.4%, and vancomycin-100%. CONCLUSION: Gram-positive bacteria were the major causes of infectious endophthalmitis in this large series, usually following surgery. CoNS was the most common isolate. Of interest, susceptibility to oxacillin and fourth-generation quinolones was lower than previously published.