Significance of large nuclear vacuoles in human spermatozoa: implications for ICSI

The aim of this study was to determine the extent of DNA fragmentation and the presence of denatured single-stranded or normal double-stranded DNA in spermatozoa with large nuclear vacuoles (LNV) selected by high magnification. Fresh semen samples from 30 patients were prepared by discontinuous isolate concentration gradient. Spermatozoa with normal nucleus (NN) and LNV were selected at x8400 magnification and placed on different slides. DNA fragmentation was determined by TUNEL assay. Denatured and double-stranded DNA was identified by the acridine orange fluorescence method. DNA fragmentation in spermatozoa with LNV (29.1%) was significantly higher (P < 0.001) than in spermatozoa with NN (15.9%). Therefore, cleavage of genomic DNA in low molecular weight DNA fragments (mono- and oligonucleosomes), and single-strand breaks (nicks) in high molecular weight DNA occur more frequently in spermatozoa with LNV. Similarly, the percentage of denatured-stranded DNA in spermatozoa with LNV (67.9%) was significantly higher (P < 0.0001) than in spermatozoa with NN (33.1%). The high level of denatured DNA in spermatozoa with LNV suggests precocious decondensation and disaggregation of sperm chromatin fibres. The results show an association between LNV and DNA damage in spermatozoa, and support the routine morphological selection and injection of motile spermatozoa at high magnification for ICSI.

Blink reflex: comparison of latency measurements in different human races

Latências do reflexo trigêmino-facial e índices cefalométricos foram analisados em 30 voluntários adultos normais, de 3 diferentes raças, sendo 10 brancos, 10 negros e 10 orientais. Idades variaram de 15 a 59 anos, alturas de 1,6 a 1,8 m e pesos de 60 a 80 kg. Os reflexos trigêmino-faciais foram obtidos por estimulação elétrica unilateral do nervo supra-orbital e captação nos músculos orbicularis oculi, para análise quantitativa de 3 respostas, ipsolateral precoce (R1), ipsolateral tardia (R2i) e contralateral tardia (R2c). Índices cefalométricos foram obtidos multiplicando-se por 100 a razão entre maior diâmetro transverso e maior diâmetro sagital do crânio. As médias dos índices cefalométricos de cada grupo foram compatíveis com as respectivas características raciais. As respostas R1, R2i e R2c não mostraram diferenças de latências estatisticamente significativas entre as 3 diferentes raças analisadas neste estudo.

Clinical Evaluation of the Effectiveness of Different Bleaching Therapies in Vital Teeth

The aims of this in vivo study were to compare the effectiveness and color stability of at-home and in-office bleaching techniques and to evaluate whether the use of light sources can alter bleaching results. According to preestablished criteria, 40 patients were selected and randomly divided into four groups according to bleaching treatment: (1) at-home bleaching with 10% carbamide peroxide, (2) in-office bleaching with 35% hydrogen peroxide (HP) without a light source, (3) in-office bleaching with 35% HP with quartz-tungsten-halogen light, and (4) in-office bleaching with 35% HP with a light-emitting diode/laser. Tooth shade was evaluated using the VITA Classical Shade Guide before bleaching as well as after the first and third weeks of bleaching. Tooth shade was evaluated again using the same guide 1 and 6 months after the completion of treatment. The shade guide was arranged to yield scores that were used for statistical comparison. Statistical analysis using the Kruskal-Wallis test showed no significant differences among the groups for any time point (P > .01). There was no color rebound in any of the groups. The bleaching techniques tested were equally effective. Light sources are unnecessary to bleach teeth. (Int J Periodontics Restorative Dent 2012;32:303-309.)

Electrical and mechanical effects of hyoscine butylbromide on the human stomach: a non-invasive approach

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

ROLE of TLR2 and TLR4 on human neutrophil functions against Paracoccidioides brasiliensis

In paracoccidioidomycosis, a systemic mycosis caused by the fungus Paracoccidioides brasiliensis (Pb), studies have focused on the role of neutrophils that are involved in the primary response to the fungus. Neutrophil functions are regulated by pro- and anti-inflammatory cytokines. Molecular mechanisms involved in this process are not fully understood, but there are strong evidences about the involvement of toll-like receptors (TLRs). We aimed at evaluating TLR2 and TLR4 expression on human neutrophils activated by GM-CSF, IL-15, TNF-alpha or IFNgamma and challenged with a virulent strain of P. brasiliensis (Pb18). Moreover, we asked if these receptors have a role on fungicidal activity, H(2)O(2) and IL-6, IL-8, TNFalpha and IL-10 production, by activating and challenging cells. All cytokines increased TLR2 and TLR4 expression. Pb18 also increased TLR2 expression, inducing an additional cytokine effect. on the contrary, it inhibited TLR4 expression. All cytokines increased neutrophil fungicidal activity and H(2)O(2) production; however, this process was not associated with TLR2 or TLR4. Neutrophil activation by GMCSF and TNF-alpha resulted in a significant increase of IL-8 production, while IL-15 and IFN-alpha have no effect. Pb18 also augmented IL-8 expression, inducing an additional effect to that of cytokines. None of the cytokines activated neutrophils by releasing IL-10. This cytokine was only detected after Pb18 challenge. Interestingly, IL-8 and IL-10 production involved TLR2 and mainly TLR4 modulation. The present results suggest that Pb18 interaction with neutrophils through TLR2 and TLR4 with consequent IL-8 and IL-10 production may be considered a pathogenic mechanism in paracoccidioidomycosis.

In vitro Cytotoxic Effect of Brazilian Green Propolis on Human Laryngeal Epidermoid Carcinoma (HEp-2) Cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Role of TLR2 and TLR4 in Human Neutrophil Functions Against Paracoccidioides brasiliensis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Description of an injury in a human caused by a false tocandira (Dinoponera gigantea, Perty, 1833) with a revision on folkloric, pharmacological and clinical aspects of the giant ants of the genera Paraponera and Dinoponera (sub-family Ponerinae)

A partir de um acidente causado pela picada de uma formiga falsa tocandira na mão de um pescador amador, os autores descrevem os achados clínicos locais observados, tais como edema, eritema e dor excruciante e a evolução do envenenamento, que cursou com fenômenos sistêmicos imediatos, como sudorese fria, náuseas, vômitos, mal estar, taquicardia e linfadenopatia axilar à esquerda. Após três horas, a dor intensa persistia e o paciente apresentou um episódio de hematoquesia, sem história anterior de enfermidades do trato digestivo, hematológicas ou vasculares. O uso de analgésicos (Tramal® 300 mg/dia), água quente e gelo não melhorou a dor, que arrefeceu em oito horas, tendo permanecido por cerca de 24 horas. São apresentados ainda os aspectos folclóricos, farmacológicos e clínicos relacionados às picadas de tocandiras.

LOSS OF THE SEX-CHROMOSOME IN HUMAN NEOPLASIAS OF THE NERVOUS-SYSTEM

Thirty-eight tumors (five grade I-II astrocytomas, three grade III astrocytomas, four glioblastomas, one oligodendroglioma, four ependymomas, one pineocytoma, three medulloblastomas, four acoustic nerve neurinomas, one intraspinal neurinoma, one neurofibroma, 10 meningiomas, and one craniopharyngioma) and three benign lesions of the nervous system were evaluated cytogenetically after in vitro culture. Sex chromosome loss was detected in 56% of the cases (-X in 13 of the 25 female patients and -Y in nine of the 16 male patients). The objective of the present report was to study the role of this abnormality in cells of the nervous system.

SUPPORT OF PARACOCCIDIOIDES-BRASILIENSIS MULTIPLICATION BY HUMAN MONOCYTES OR MACROPHAGES - INHIBITION BY ACTIVATED PHAGOCYTES

The interaction of human monocytes or monocyte-derived macrophages and yeast-form Paracoccidioides brasiliensis was studied in vitro. Yeast cells were readily ingested by adherent monocytes or macrophages. Multiplication of P. brasiliensis, measured by growth as colony forming units (cfu) on a supplemented medium with good plating efficiency, was greater in monocyte co-cultures compared to the number of cfu obtained from complete tissue-culture medium (CTCM). Multiplication increased with time in macrophage cocultures, e.g., from two-six-fold in 24 h to nine-fold in 72 h. Microscopic observations indicated that ingested yeast cells multiplied inside macrophages. When monocytes were treated with supernate cytokines (CK) from concanavalin-A-stimulated mononuclear cells, then co-cultured with P. brasiliensis, multiplication was significantly inhibited compared with control monocyte co-cultures. Treatment of macrophages-derived from monocytes by culture in vitro for 3 days-for a further 3 days with CK resulted in maximal inhibition of multiplication over the subsequent 72 h. Similarly, when monocyte-derived macrophages (after culture for 7 days) were treated for 3 days with recombinant human gamma-interferon (IFN; 300 U/ml) or CK they restricted multiplication of P. brasiliensis by 65% and 95%, respectively, compared with control macrophages, Antibody to IFN abrogated the effect of IFN or CK treatment. These findings show that ingested P. brasiliensis can multiply in human monocytes or macrophages and that this multiplication can be restricted by activated monocytes or macrophages.