Supplemental progesterone during early pregnancy exerts divergent responses on embryonic characteristics in sows and gilts.

Progesterone (P4) plays a key role in pregnancy establishment and maintenance; during early pregnancy, P4 stimulates the production and release of uterine secretions necessary for conceptus growth prior to implantation; therefore, exogenous P4 supplementation may improve embryo development. This study evaluated the effects of supplementation during early pregnancy with long-acting injectable progesterone or altrenogest on embryonic characteristics of sows and gilts. Thus, a total of 32 sows and 16 gilts were used. On day 6 of pregnancy sows and gilts were allocated to one of the following groups: non-supplemented; supplemented with 20 mg of altrenogest, orally, from days 6 to 12 of pregnancy; supplemented with 2.15 mg/kg of long-acting injectable progesterone on day 6 of pregnancy. Animals were killed on day 28 of pregnancy, and ovulation rate, embryo survival, embryo weight, crown-to-rump length, uterine glandular epithelium and endometrial vascularization were assessed. Treatments had no effect on pregnancy rate, embryo survival or endometrial vascular density (P > 0.05). Non-supplemented gilts presented larger and heavier embryos compared to gilts from supplemented groups (P < 0.05). Sows in the altrenogest group presented larger and heavier embryos compared to non-supplemented sows and sows supplemented with long-acting injectable progesterone. In conclusion, supplementation of sows and gilts with progestagen from day 6 of pregnancy can be used as a means to improve embryo survival without deleterious effects.

Absence of seminal plasma from sperm-rich fraction decreases boar sperm quality characteristics during the course of liquid storage.

Seminal plasma (SP), the fluid that surrounds the sperm cells, is known to exert substantial influence on sperm physiology. The SP has a pivotal role in sperm function in vivo, and due to its components, it functions in an ambiguous manner in vitro, simultaneously possessing deleterious and beneficial effects. This experiment aimed to describe the differences between the presence or absence of SP from the sperm-rich fraction on some spermatozoa characteristics (kinetics, plasma and acrosome membrane integrity, lipid peroxidation and capacitation-like changes). Furthermore, this experiment focused on distinguishing the effects of SP on the variables evaluated from the effects of centrifugation during SP removal. Total and progressive sperm motility, as well as integrity of plasma and acrosome membranes, were less (P < 0.05) in the absence of SP. Membrane lipid peroxidation (P < 0.05) and sperm membrane stability (P < 0.05) did not differ among treatments. The SP from the sperm-rich fraction is important for the maintenance of adequate structural and functional characteristics of extended liquid boar semen and should be present in seminal doses throughout storage. Furthermore, the detrimental effect on the variables evaluated was caused solely by the absence of SP and not by the process of removal through centrifugation at 500 x g for 10 min.

Seminal plasma arising from the whole boar sperm-rich fraction increases the stability of sperm membrane after thawing.

Boar spermatozoa arising from the sperm-rich ejaculate fraction are reported to have a more stable plasma membrane and are more resistant to cold shock and premature acrosome reaction than spermatozoa from the whole ejaculate. Furthermore, seminal plasma (SP) can increase the cryotolerance of boar spermatozoa, and in other domestic species, it has the ability to reverse cryopreservation damage. This study aimed to evaluate the effects of boar SP arising from the whole sperm-rich ejaculate fraction (SP-SRF) on the integrity, stability, and peroxidation of sperm membranes after thawing. Each ejaculate ( = 24) was divided among 4 treatments: control (CT), centrifuged and suspended in autologous SP-SRF (CS), centrifuged with withdrawn SP-SRF (CW), and post-thawed SP arising from the whole sperm-rich fraction addition to CW (CWSP). After thawing, all treatments were incubated for 5, 60, and 120 min and were analyzed for membrane integrity, fluidity, and peroxidation by flow cytometer. The absence of SP-SRF increased the lipid disorder ( < 0.05) but had no effect on lipid peroxidation ( > 0.05) or membrane integrity ( > 0.05). However, the increase in lipid disorder by withdrawal of SP-SRF was reversed by SP-SRF addition ( < 0.05) to the post-thawing medium, whereas plasma and acrosomal membrane integrity ( > 0.05) and lipid peroxidation ( > 0.05) were unchanged. In conclusion, despite the centrifugation effects, the addition of SP arising from the whole sperm-rich fraction to post-thawed boar semen decreased sperm lipid disorder without an influence of the sperm membrane integrity and peroxidation.

Organic selenium supplementation increases PHGPx but does not improve viability in chilled boar semen.

This study evaluated the effects of dietary organic selenium (Se) on viability of chilled boar semen. Twelve boars were divided into three groups: control (CON), 0.3 mg kg(-1) sodium selenite; inorganic (INO), 0.5 mg kg(-1) sodium selenite and organic (ORG), 0.5 mg kg(-1) Se yeast. The experiment was conducted within 10 weeks, and analysis was performed fortnightly, in storage semen by 72 h. No effect was observed on motility; however, straightness and linearity percentages were higher (P < 0.05) in the animals receiving CON diet compared with INO group. Percentages of cells with both plasma and acrosomal intact membranes, lipidic membrane peroxidation and mitochondrial membrane potential were similar on all treatments. Animals receiving CON diet presented higher (P < 0.05) values of ATP when compared with INO group. The PHGPx was higher (P < 0.05) in animals that received ORG in comparison with INO group. In conclusion, organic selenium supplementation increases PHGPx but does not improve chilled semen viability in 72 h.

Gonadotropin-induced puberty does not impair reproductive performance of gilts over three parities.

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty-nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post-weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning-to-oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.

Effect of sequence of insemination after simultaneous thawing of multiple semen straws on conception rate to timed AI in suckled multiparous Nelore cows.

The objective was to determine the effect of sequence of insemination after simultaneous thawing of multiple 0.5 mL semen straws on conception rate in suckled multiparous Nelore cows. The effect of this thawing procedure on in vitro sperm characteristics was also evaluated. All cows (N = 944) received the same timed AI protocol. Ten straws (0.5 mL) of frozen semen from the same batch were simultaneously thawed at 36 °C, for a minimum of 30 sec. One straw per cow was used for timed AI. Frozen semen from three Angus bulls was used. Timed AI records included sequence of insemination (first to tenth) and time of semen removal from thawing bath. For laboratory analyses, the same semen batches used in the field experiment were evaluated. Ten frozen straws from the same batch were thawed simultaneously in a thawing unit identical to that used in the field experiment. The following sperm characteristics were analyzed: sperm motility parameters, sperm thermal resistance, plasma and acrosomal membrane integrity, lipid peroxidation, chromatin structure, and sperm morphometry. Based on logistic regression, there were no significant effects of breeding group, body condition score, AI technician, and sire on conception rate, but there was an interaction between sire and straw group (P = 0.002). Semen from only one bull had decreased (P < 0.05) field fertility for the group of straws associated with the longest interval from thawing to AI. However, the results of the laboratory experiment were unable to explain the findings of the field experiment. Sperm width:length ratio of morphometric analysis was the single sperm characteristic with a significant interaction between sire and straw group (P = 0.02). It was concluded that sequence of insemination after simultaneous thawing of 10 semen straws can differently affect conception rates at timed AI, depending on the sire used. Nevertheless, the effects of this thawing environment on in vitro sperm characteristics, remain to be further investigated.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association.

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Addition of seminal plasma to post-thawing equine semen: what is the effect on sperm cell viability?

Effect of seminal plasma addition after thawing on viability or cryocapacitation is not definitively established. This experiment was performed to verify the effect of adding seminal plasma, autologous or homologous (from an animal with good semen freezability). Five ejaculates from each of four stallions with proven fertility were collected and cryopreserved. The semen was subsequently thawed and divided into the following three treatment groups: no seminal plasma addition after semen thawing (NOSP); the addition of homologous seminal plasma after semen thawing (HSP) and the addition of autologous seminal plasma after semen thawing (ASP). The addition of 20% of seminal plasma led to an increase in the cell population that simultaneously show plasma and acrosomal membrane integrity (p < 0.05). The addition of seminal plasma did not alter the total motility, the amount of cells with mitochondrial membrane potential or the sperm velocities (average path velocity, straight-line velocity and curvilinear velocity). However, the beat/cross-frequency, straightness and linearity were reduced in ASP and HSP groups (p < 0.05). Unexpectedly, the addition of homologous seminal plasma reduced the proportion of cells with progressive motility (p < 0.05) and the addition of autologous seminal plasma reduced the amplitude of the lateral head displacement (p < 0.05). Based on the increase in the cell populations that had the plasma and acrosomal membrane integrity simultaneously identified in this study, we proposed that the addition of seminal plasma (autologous or homologous) into post-thawed semen before insemination could increase semen fertility.

[Intracranial pressure monitoring in children with fulminant hepatic failure]. / Monitorizacion de la presion intracraneal en ninos con insuficiencia hepatica fulminante.

INTRODUCTION: We describe a series of 10 children with intracranial hypertension complicating fulminant hepatic failure submitted to intracranial pressure (ICP) monitoring for intensive care and transplantation management. PATIENTS AND METHODS: Information from pediatrics patients acute liver failure admitted to our hospital was collected in a standard protocol form. We analyzed data from 10 patients, medium age 5.2 years old. In this period we studied aspects as ICP transducer used, number of days with ICP monitoring and complications of ICP monitoring. RESULTS: Hepatitis A was diagnosed in five patients and hepatitis B in two cases. The initial ICP were 2 to 24 mmHg in transducer. Seven patients died, four due to intracranial hypertension, included the patient operated for subdural hematoma, and three with transplantation failure. Only a case of hematoma was verified. CONCLUSIONS: The application of ICP monitoring allows intensive care for aggressive ICP management. It can be used in children without adaptations.

Practical techniques for bovine sperm simultaneous fluorimetric assessment of plasma, acrosomal and mitochondrial membranes.

This experiment was performed to develop and validate practical techniques for simultaneous evaluation of the integrity of plasma and acrosomal membranes, as well as mitochondrial function in bovine spermatozoa using associations of fluorescent probes. Four protocols of fluorescent probes association were defined: protocol 1: propidium iodide (PI), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and rhodamine 123; protocol 2: PI, FITC-PSA and MitoTracker Green FM (MITO); protocol 3: PI, Hoechst 33342 (H342), FITC-PSA and CMXRos; and protocol 4: PI, H342, FITC-PSA and JC-1. Three ejaculates from each of the four bulls (n = 12) were utilized, showing sperm motility >/=80% and abnormal morphology

Fluorescent stain method for the simultaneous determination of mitochondrial potential and integrity of plasma and acrosomal membranes in boar sperm.

The purpose of this study was to validate a technique for simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in boar spermatozoa, using an association of fluorescent probes: Propidium iodide (PI), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and JC-1. Three ejaculates from each of four different boars, all showing motility >or=80% and abnormal morphology

Estudo comparativo entre os sistemas de umidificação aquoso aquecido e trocador de calor e de umidade na via aérea artificial de pacientes em ventilação mecânica invasiva / Comparative study between heated water humidifiers and hygroscopic heat and moisture exchangers in artificial airways for patients undergoing invasive mechanical ventilation

CONTEXTO: Nos pacientes recebendo suporte ventilatório invasivo através de tubos endotraqueais é essencial o uso de umidificadores. OBJETIVO: avaliar os níveis de temperatura (TºC) e de umidade relativa (UR) do gás administrado ao paciente em ventilação mecânica através da umidificação aquosa aquecida (UAA) e do filtro trocador de calor e de umidade (FTCU). MÉTODO: Este foi um estudo prospectivo, randomizado, onde foram estudados 20 pacientes divididos em dois grupos: um grupo usou a UAA (n=10) e o outro grupo (n=10) usou FTCU Hygrobac "S", marca Mallinckrodt®. As variáveis analisadas foram: níveis de temperatura (T) e umidade relativa (UR) do gás, volume minuto (VM), volume corrente (VC) e volume de condensação. RESULTADOS: Verificou-se que o sistema de umidificação aquosa aquecida atingiu temperaturas mais baixas que o filtro trocador (29,01 ± 1,33 ºC, versus 30,14 ± 1,24 ºC; p<0,001). A umidade relativa foi maior na UAA do que no FTCU (97,45 ± 5,22 por cento, versus 89,87 ± 11.04 por cento, p < 0,021). O volume de condensação do circuito do grupo usando UAA foi maior que o do grupo usando o FTCU (p < 0,05). CONCLUSÃO: Os resultados demonstram que ambos os sistemas, UAA e o FTCU forneceram umidades absolutas abaixo do valor recomendado, sendo que o sistema de UAA ofereceu uma umidade relativa maior, enquanto que o aquecimento do gás com o FTCU obteve um melhor desempenho.

BACKGROUND: In patients receiving invasive mechanical ventilation through endotracheal tubes, the use of humidifiers is essential. OBJECTIVE: To evaluate temperature and relative humidity levels in the gas administered to patients undergoing mechanical ventilation by means of heated water humidifiers (HWH) and hygroscopic heat and moisture exchangers (HHME). METHOD: This was a prospective randomized study on 20 patients divided into two groups: one group using HWH (n=10) and the other using the Hygrobac "S" model of HHME, made by Mallinckrodt® (n=10). The variables analyzed were: temperature and relative humidity levels of the gas, minute volume (MV), tidal volume (V T) and condensation volume. RESULTS: It was found that HWH attained lower temperatures than did HHME (29.01 ± 1.33 ºC versus 30.14 ± 1.24 ºC; p<0.001). The relative humidity was higher in HWH than in HHME (97.45 ± 5.22 percent versus 89.87 ± 11.04 percent; p<0.021). The condensation volume in the ventilator circuit for the group using HWH was greater than for the HHME group (p<0.05). CONCLUSION: The results demonstrate that both systems (HWH and HHME) supplied absolute humidity that was below recommended values, while the HWH offered higher relative humidity. On the other hand, HHME produced better performance regarding gas heating.

Guidelines for neurosurgical trauma in Brazil.

This chapter emphasizes some aspects of the Brazilian Guidelines for the Assessment of Head Injury Patients, written based on the experience of the Emergency Service, Neurosurgical Division of the University of São Paulo Medical School Hospital, and sponsored by the Brazilian Society of Neurosurgery. These guidelines approach the management of head-injury patients from their initial assessment in the Emergency Room until the final suggested clinical or surgical management. The Brazilian Guidelines represents our efforts to provide the basis for a common unified data collection system, which may allow cooperative studies in the future.

Spontaneous migration of a bullet in the cerebellum--case report.

A 15-year-old boy presented with a gunshot wound in the left cerebellar hemisphere. He was confused and left cerebellar signs were noted. The patient underwent the first surgery for debridement of the entry wound in the left parietal region and second surgery to remove the bullet. However, the bullet could not be located via a left unilateral suboccipital craniectomy in the park bench position, because it had migrated to the opposite side due to the effects of gravity in just a few hours. Skull radiography obtained just before the third surgery showed that the bullet had returned to the left side, and it was removed easily via the previous craniectomy in the sitting position. The clinical course suggests that in removing a bullet, skull radiography or computed tomography should be obtained just before surgery, or even intraoperatively, and that those findings should be the basis for the surgical procedure and operative position.

Recovery from Duret hemorrhage: a rare complication after craniotomy--case report.

A 44-year-old female presented with Duret hemorrhage due to transtentorial herniation by extradural hematoma as a complication after craniotomy for treatment of spontaneous middle cranial fossa cerebrospinal fluid leakage through the oval window. Brain computed tomography revealed linear hemorrhage in the midbrain and the rostral pons. She awoke after 2 weeks in a coma, despite showing ocular bobbing and bilateral intranuclear ophthalmoplegia. She was discharged from the hospital with minimal neurological defects. Duret hemorrhage is usually fatal, but this case shows that early surgical decompression is the most important factor to avoid the worst sequelae.

Traumatic acute giant epidural hematoma in a hydrocephalic shunted child.

Extradural hematoma (EDH) is considered to be a rare complication of head trauma in children, and represents a serious and urgent pathology from which complete recovery can be expected if specialized treatment is instituted in time. In this article, the authors report the potential danger to a hydrocephalic shunted child who was apparently asymptomatic at the time of hospital admission with a mild head injury and developed an EDH of venous origin. This child had a rapid (time interval from injury to decerebrate posture of about 2 h), atypical (remained asymptomatic most of the time until abruptly deterioration) and fatal course, stressing the importance of early diagnosis and rapid therapy in order to avoid the death of the patient. The authors discuss the role of the ventriculoperitoneal shunting system in the lack of clinical symptoms associated with the presence of a giant EDH and a rapid and fatal course, and stress the importance of computed tomographic (CT) scanning in these patients, even if they are asymptomatic. If a skull fracture is suspected, a CT scan must be performed without delay.

Spontaneous epidural hematoma following a shunt in an infant with congenital factor X deficiency. Case report and literature review.

The authors describe a case of an infant with congenital factor X deficiency. The patient presented with a central nervous system hemorrhage followed by hydrocephalus. He underwent a ventriculoperitoneal shunt and, during the postoperative period, developed a spontaneous epidural hematoma, which was evacuated. The clinical and pathophysiological aspects of this case are discussed based on a literature review.

Bacteroides fragilis: an exogenous pathogen?

Thirteen strains of Bacteroides fragilis isolated from contaminated water or from cases of intestinal and non-intestinal infections were comparatively analyzed in order to detect a possible biological relationship among them. The observation of similar profiles among the two groups, regarding the capacity of inducing abscesses and some surface properties, may support the hypothesis that the species could also act as an exogenous pathogen.

Metabolic and morphological changes in A. albopictus cells infected with Mayaro virus under heat-shock conditions.

We addressed the question how temperature elevation inhibits Mayaro virus replication in Aedes albopictus infected cells. The morphology and macromolecular changes induced by temperature, infection and high serum concentration were investigated in these cells. Cells incubated with 2 and 10% serum at 28 degrees C disclosed an intense vacuolization and inhibition of [35S]methionine incorporation in a time-dependent manner. 34 and 50 kDa viral structural proteins were detected 24 h after infection. In contrast, an inhibition of viral proteins synthesis occurred when infected cells were kept at 37 degrees C (heat-shock conditions). Total cellular RNA was isolated from mock and infected cells incubated at 28 or 37 degrees C. Northern blot analysis with a Mayaro genomic probe coding for viral structural proteins showed a decrease in the amount of viral 26S RNA in stressed cells when compared to those kept at 28 degrees C. Taken together, these results suggest that the inhibition of viral proteins synthesis in response to temperature elevation is associated with a decrease in the amount of subgenomic 26S RNA.

Changes in cell-surface carbohydrates of Trypanosoma cruzi during metacyclogenesis under chemically defined conditions.

Highly purified lectins with specificities for receptor molecules containing sialic acid, N-acetylglucosamine (D-GlcNAc), N-acetylgalactosamine (D-GalNAc), galactose (D-Gal), mannose-like residues (D-Man) or L-fucose (L-Fuc), were used to determine changes in cell-surface carbohydrates of the protozoal parasite Trypanosoma cruzi during metacyclogenesis under chemically defined conditions. Of the D-GalNAc-binding lectins, BS-I selectively agglutinated metacyclic trypomastigotes, MPL was selective for replicating epimastigotes, whereas SBA strongly agglutinated all developmental stages of T. cruzi. WGA (sialic acid and/or D-GlcNAc specific) was also reactive with differentiating epimastigotes and metacyclic trypomastigotes but displayed a higher reactivity with replicating epimastigote forms. A progressive decrease in agglutinating activity was observed for jacaline (specific for D-Gal) during the metacyclogenesis process; conversely, a progressive increase in affinity was observed for RCA-I (D-Gal-specific), although the reactivity of other D-Gal-specific lectins (PNA and AxP) was strong at all developmental stages. All developmental stages of T. cruzi were agglutinated by Con A and Lens culinaris lectins (specific for D-Man-like residues); however, they were unreactive with the L-fucose-binding lectins from Lotus tetragonolobos and Ulex europaeus. These agglutination assays were further confirmed by binding studies using 125I-labelled lectins. Neuraminidase activity was detected in supernatants of cell-free differentiation medium using the PNA hemagglutination test with human A erythrocytes. The most pronounced differences in lectin agglutination activity were observed between replicating and differentiating epimastigotes, suggesting that changes in the composition of accessible cell-surface carbohydrates precede the morphological transformation of epimastigotes into metacyclic trypomastigotes.