Avaliação de diferentes crioprotetores e taxas de diluição na criopreservação seminal de Prochilodus brevis / Evaluation of different crioprotectants and dilution ratios on Prochilodus brevis sperm cryopreservation

Este estudo objetivou caracterizar o sêmen de Prochilodus brevis e avaliar os efeitos de diferentescrioproterores e taxas de diluição sobre a cinética e a morfologia do sêmen criopreservado. Inicialmente,amostras seminais de P. brevis (n = 40) foram analisadas quanto ao volume, pH, osmolaridade, concentração,motilidade e morfologia espermáticas. Para a criopreservação, o material coletado foi distribuído em 10 pools desêmen (n = 10), submetidos a seis tratamentos compostos pela combinação de glicose 5%, dois crioprotetores(DMSO ou MG) e três taxas de diluição (1:3, 1:6 ou 1:9 sêmen:diluidor). As amostras, in natura e pósdescongeladas,foram analisadas quanto à sua morfologia e cinética espermática utilizando o CASA. O sêmen deP. brevis apresentou características semelhantes a demais espécies de Prochilodus. O sêmen congelado comglicose e MG apresentou resultados significativamente superiores (P < 0,05) comparado àquele congeladoutilizando DMSO. As diluições 1:3 e 1:6 apresentaram melhores resultados para glicose + MG, enquanto 1:9 foimelhor para glicose + DMSO. Portanto, a associação glicose + MG, numa taxa de diluição de 1:3 ou 1:6, é amais indicada para a criopreservação do sêmen de Prochilodus brevis.(AU)

This study aimed to characterize Prochilodus brevis sperm and evaluate the effects of different diluentsand dilution ratios on kinetics and morphology of cryopreserved sperm. Initially, sperm samples of P. brevis(n = 40) were assessed for volume, pH, osmolality, spermatic concentration, motility and morphology. Forcryopreservation, the collected material was distributed in ten pools of semen (n = 10), submitted to sixtreatments composed by a combination of glucose 5% with two cryoprotectants (DMSO or MG) and threedilution ratios (1:3, 1:6 or 1:9 - sperm:diluent). In natura and post-thawed samples were assessed for spermaticmorphology and kinetics using CASA. The sperm of P. brevis showed similar characteristics to otherProchilodus species. Sperm cryopreserved in glucose plus MG presented significantly higher results (P < 0.05)compared to sperm frozen in DMSO. The dilution ratios of 1:3 and 1:6 yielded better results for glucose + MG,while 1:9 was the best dilution for glucose + DMSO. In conclusion, the association of glucose + MG, in adilution ratio of 1:3 or 1:6, is the most indicated for Prochilodus brevis sperm cryopreservation.(AU)

Avaliação de diferentes crioprotetores e taxas de diluição na criopreservação seminal de Prochilodus brevis / Evaluation of different crioprotectants and dilution ratios on Prochilodus brevis sperm cryopreservation

Este estudo objetivou caracterizar o sêmen de Prochilodus brevis e avaliar os efeitos de diferentescrioproterores e taxas de diluição sobre a cinética e a morfologia do sêmen criopreservado. Inicialmente,amostras seminais de P. brevis (n = 40) foram analisadas quanto ao volume, pH, osmolaridade, concentração,motilidade e morfologia espermáticas. Para a criopreservação, o material coletado foi distribuído em 10 pools desêmen (n = 10), submetidos a seis tratamentos compostos pela combinação de glicose 5%, dois crioprotetores(DMSO ou MG) e três taxas de diluição (1:3, 1:6 ou 1:9 – sêmen:diluidor). As amostras, in natura e pósdescongeladas,foram analisadas quanto à sua morfologia e cinética espermática utilizando o CASA. O sêmen deP. brevis apresentou características semelhantes a demais espécies de Prochilodus. O sêmen congelado comglicose e MG apresentou resultados significativamente superiores (P < 0,05) comparado àquele congeladoutilizando DMSO. As diluições 1:3 e 1:6 apresentaram melhores resultados para glicose + MG, enquanto 1:9 foimelhor para glicose + DMSO. Portanto, a associação glicose + MG, numa taxa de diluição de 1:3 ou 1:6, é amais indicada para a criopreservação do sêmen de Prochilodus brevis.

This study aimed to characterize Prochilodus brevis sperm and evaluate the effects of different diluentsand dilution ratios on kinetics and morphology of cryopreserved sperm. Initially, sperm samples of P. brevis(n = 40) were assessed for volume, pH, osmolality, spermatic concentration, motility and morphology. Forcryopreservation, the collected material was distributed in ten pools of semen (n = 10), submitted to sixtreatments composed by a combination of glucose 5% with two cryoprotectants (DMSO or MG) and threedilution ratios (1:3, 1:6 or 1:9 - sperm:diluent). In natura and post-thawed samples were assessed for spermaticmorphology and kinetics using CASA. The sperm of P. brevis showed similar characteristics to otherProchilodus species. Sperm cryopreserved in glucose plus MG presented significantly higher results (P < 0.05)compared to sperm frozen in DMSO. The dilution ratios of 1:3 and 1:6 yielded better results for glucose + MG,while 1:9 was the best dilution for glucose + DMSO. In conclusion, the association of glucose + MG, in adilution ratio of 1:3 or 1:6, is the most indicated for Prochilodus brevis sperm cryopreservation.

TP53 codon 72 polymorphism in adult soft tissue sarcomas.

Soft tissue sarcomas (STS) are tumors of mesodermal origin, comprising about 1% of all adult neoplasms. Management of such tumors is an important medical challenge. TP53 codon 72 polymorphism results in either the arginine or proline form of the p53 protein; several studies have investigated whether codon 72 polymorphisms are risk and prognostic factors for cancer. We investigated p53 codon 72 polymorphism (Arg72Pro) frequencies with respect to the susceptibility and the clinical outcome of patients with STS. A series of 100 STS were genotyped for the p53 Arg72Pro polymorphism using polymerase chain reaction. Genotype frequencies were compared to a group of 85 healthy donors (controls). Possible associations between polymorphic genotypes, clinicopathological factors and survival of STS patients were also investigated. Genotypic frequencies obtained for STS patients did not significantly differ from that obtained for controls. In the STS group, p53 codon 72 polymorphic variants were not significantly associated with gender, age, tumor size, clinical stage, tumor grade, histology, or nodal or distant metastasis. The five-year overall survival rate for the STS group was 48%; it was significantly affected by tumor grade, clinical stage, and nodal and distant metastasis. Soft tissue sarcoma patients with the Pro/Pro variant had a reduced survival rate (30%), when compared to the p53 Arg/Arg (45%) and the p53 Arg/Pro groups (55%). However, the differences between these groups were not significant (P = 0.44).

[Chronic intestinal pseudo-obstruction in children - report of seven cases] / Pseudo-obstrução intestinal crônica na infância - relato de sete casos.

OBJECTIVE: The purpose of this report is to describe the clinical characteristics and histological abnormalities on full-thickness intestinal biopsy in seven children from the Gastroenterology (GI) outpatient clinic. METHODS: A retrospective study of all patients who attended the GI outpatient clinic in the last ten years. The medical records of seven patients were reviewed, and all the relevant clinical data recorded. Diagnosis was based on clinical, laboratorial, and histological findings. CONCLUSION: Although this is a rare syndrome, the differential diagnosis shall be performed whenever the patient presents signs and symptoms of obstruction along the digestive tract with no association of any mechanical cause. Studies shall be encouraged regarding the precocious and accurate diagnosis, using less invasive techniques and specific therapeutics (both clinical and surgical), in order to improve life quality and extended survival of affected patients.

Ethological analysis of mother-pup interactions and other behavioral reactions in rats: effects of malnutrition and tactile stimulation of the pups.

Mother-pup interaction, as well as other behavioral reactions were studied during the lactation period in 24 litters of Wistar rats and their dams fed either a 16% (control - C; 12 litters) or a 6% (malnourished - M; 12 litters) protein diet. The diets were isocaloric. Throughout lactation there was a 36.4% weight loss of M dams and a 63% body weight deficit in the M pups when compared to control pups. During this period, half of the litters were exposed daily to additional tactile stimulation (CS or MS), while the other half were submitted to normal rearing conditions (CN or MN). The tactile stimulation of pups (handling) consisted of holding the animal in one hand and gently touching the dorsal part of the animal's body with the fingers for 3 min. A special camera and a time-lapse video were used to record litter behavior in their home cages. Starting at 6 p.m. and ending at 6 a.m., on days 3, 6, 12, 15, 18 and 21 of lactation, photos were taken at 4-s intervals. An increase in the frequency (154.88 +/- 16.19) and duration (455.86 +/- 18.05 min) of suckling was observed throughout the lactation period in all groups compared to birth day (frequency 24.88 +/- 2.37 and duration 376.76 +/- 21.01 min), but the frequency was higher in the C (84.96 +/- 8.52) than in the M group (43.13 +/- 4.37); however, the M group (470.2 +/- 11.87 min) spent more time suckling as compared with the C group (393.67 +/- 13.09 min). The M dams showed a decreased frequency of resting position throughout the lactation period (6.5 +/- 2.48) compared to birth day (25.42 +/- 7.74). Pups from the C group were more frequently observed separated (73.02 +/- 4.38) and interacting (258.99 +/- 20.61) more with their mothers than the M pups (separated 66.94 +/- 5.5 and interacting 165.72 +/- 12.05). Tactile stimulation did not interact with diet condition, showing that the kind of stimulation used in the present study did not lead to recovery from the changes induced by protein malnutrition. The changes in mother-pup interaction produced by protein malnutrition of both may represent retardation in neuromotor development and a higher dependence of the pups on their mothers. These changes may represent an important means of energy saving and heat maintenance in malnourished pups.

Ethological analysis of mother-pup interactions and other behavioral reactions in rats: effects of malnutrition and tactile stimulation of the pups

Mother-pup interaction, as well as other behavioral reactions were studied during the lactation period in 24 litters of Wistar rats and their dams fed either a 16 percent (control - C; 12 litters) or a 6 percent (malnourished - M; 12 litters) protein diet. The diets were isocaloric. Throughout lactation there was a 36.4 percent weight loss of M dams and a 63 percent body weight deficit in the M pups when compared to control pups. During this period, half of the litters were exposed daily to additional tactile stimulation (CS or MS), while the other half were submitted to normal rearing conditions (CN or MN). The tactile stimulation of pups (handling) consisted of holding the animal in one hand and gently touching the dorsal part of the animal's body with the fingers for 3 min. A special camera and a time-lapse video were used to record litter behavior in their home cages. Starting at 6 p.m. and ending at 6 a.m., on days 3, 6, 12, 15, 18 and 21 of lactation, photos were taken at 4-s intervals. An increase in the frequency (154.88 + or - 16.19) and duration (455.86 + or - 18.05 min) of suckling was observed throughout the lactation period in all groups compared to birth day (frequency 24.88 + or - 2.37 and duration 376.76 + or - 21.01 min), but the frequency was higher in the C (84.96 +- 8.52) than in the M group (43.13 + or - 4.37); however, the M group (470.2 + or - 11.87 min) spent more time suckling as compared with the C group (393.67 + or - 13.09 min). The M dams showed a decreased frequency of resting position throughout the lactation period (6.5 + or- 2.48) compared to birth day (25.42 + or - 7.74). Pups from the C group were more frequently observed separated (73.02 + or - 4.38) and interacting (258.99 + or - 20.61) more with their mothers than the M pups (separated 66.94 + or - 5.5 and interacting 165.72 + or - 12.05). Tactile stimulation did not interact with diet condition, showing that the kind of stimulation used in the present study did not lead to recovery from the changes induced by protein malnutrition. The changes in mother-pup interaction produced by protein malnutrition of both may represent retardation in neuromotor development and a higher dependence of the pups on their mothers. These changes may represent an important means of energy saving and heat maintenance in malnourished pups

Cardiac abnormalities in the acquired immunodeficiency syndrome. A prospective study with a clinical-pathological correlation in twenty-one adult patients.

OBJECTIVE: To evaluate the cardiac abnormalities and their evolution during the course of the acquired immunodeficiency syndrome, as well as to correlate clinical and pathological data. METHODS: Twenty-one patients, admitted to the hospital with the diagnosis of acquired immunodeficiency syndrome, were prospectively studied and followed until their death. Age ranged from 19 to 42 years (17 males). ECG and echocardiogram were also obtained every six months. After death, macro- and microscopic examinations were also performed. RESULTS: The most frequent causes of referral to the hospital were: diarrhea or repeated pneumonias, tuberculosis, toxoplasmosis or Kaposi sarcoma. The most frequent findings were acute or chronic pericarditis (42%) and dilated cardiomyopathy (19%). Four patients died of cardiac problems: infective endocarditis, pericarditis with pericardial effusion, bacterial myocarditis and infection by Toxoplasma gondii. CONCLUSION: Severe cardiac abnormalities were the cause of death in some patients. In the majority of the patients, a good correlation existed between clinical and anatomical-pathological data. Cardiac evaluation was important to detect early manifestations and treat them accordingly, even in asymptomatic patients.

Effects of Different Levels of Protein and Environmental Stimulation on the Behavior of Young Rats Tested in the Elevated Plus-Maze.

UNLABELLED: The interaction between the effects of different levels of protein malnutrition and environmental stimulation was investigated in young male rats in the elevated plus-maze (EPM). Litters (dam plus 6 male and 2 female pups) were fed a 16% ( CONTROL: C), 10% (Malnourished: M10) or 6% (Malnourished: M6) protein diet. Half of the litters were daily exposed to additional stimulation (CS, MS10 or MS6), while the other half was maintained in normal rearing conditions (CN, MN10 or MN6). The stimulation (handling) consisted of holding the rat in one hand and gently touching the dorsal part of the body with the fingers for 3 min. On the 22nd day of life (weaning), two male pups from each group were tested in the plus-maze. Two male pups continued to receive the same diet as their mothers, and the other two were fed a non-purified lab chow diet until 35 days of age when they were tested in the plus-maze (LN, LS, LN10, LS10, LN6 and LS6 groups). Results showed that environmental stimulation increased open arms exploration both at 22 and 35 days of age, indicating an anxiolytic effect of this procedure. Younger rats (22 days of age) explored significantly more the open arms of the maze as compared with older rats (35 days of age), indicating an increase in anxiety with age. M6 animals showed significantly higher percentages of open arm entries and less frequent attempts to enter open arms in the maze as compared with C and M10 animals. These results suggest that even a short period of protein deficiency can produce alterations in the emotional response of rats in the elevated plus-maze. In addition, the data demonstrated that protein deficiency more severe than 10% is necessary to produce behavior alterations in the EPM test.

[Cardiac changes in children with AIDS]. / Alterações cardíacas em crianças com SIDA.

PURPOSE: To describe the cardiac findings in children with AIDS, the follow-up with treatment and the correlation between clinical and pathological features. METHODS: We studied prospectively 25 children with clinical-laboratorial diagnosis of AIDS, ages between 3 months and 11 years, even those without cardiac symptoms. We classified the signs of AIDS following that of CDC-Atlanta 1994. Eight children died and it was done necropsy in six with macro and microscopic examinations. RESULTS: Fifteen cases had already complications of AIDS and were classified as C2 and C3, 5 as B2, 3 B1 and 2 as A1. Beside the symptoms related to the disease and infections we found signs of congestive heart failure III and IV (NYHA) in 5 children, pericardial effusion in 5, (one of them had cardiac tamponade). In the electrocardiogram (EKG), 8 children had repolarization abnormalities. In 1st echocardiogram (ECHO) we found some features of dilated myocardiopathy in 8 (6 were asymptomatic) 5 of those had pericardial effusion, one child had huge amount of pericardial liquid and also increased measures of intraventricular septum and posterior wall of left ventricle by ECHO and confirmed by necropsy examination. All cases of diagnosis of myocardiopathy received treatment with captopril and 4 also received furosemide and digoxina. Even under oral treatment three of them had deterioration of heart failure but after combined anti-retroviral drugs they showed better cardiac functions. From the six necropsied cases, two had increased heart weight without myocarditis, one had toxoplasma pancarditis and other one had fibrocalcic vasculopathy. CONCLUSION: We found several assymptomatic cases already with decreased ventricular function. Some worsened of heart failure even under apropriate treatment and showed better cardiac index after combined anti-retroviral drugs.

[Clinicopathologic correlation in 50 cases of acquired immunodeficiency syndrome. Retrospective study]. / Correlação clínicopatológica de 50 casos de síndrome de imunodeficência adquirida. Estudo retrospectivo.

PURPOSE: To analyze myocardial abnormalities in patients of acquired immunodeficiency syndrome with clinical and pathological correlation. METHODS: We selected 50 cases, retrospectively, age ranged from 3 months to 40 years, all of them had myocardial changes and the data of clinical records fulfilled our protocol. Cases of others cardiac diseases were not included. RESULTS: The pathological findings were: myocarditis in 33 (11 had severe myocarditis) and degenerative hystological lesions in 17. The etiologic agents detected were: Toxoplasma in 11, Cryptococcus in 7 and Cytomegalovirus in 3. In 12 cases we could not find any agent. In 15 cases occurred others lesions: endocarditis, pericarditis and sarcoma of Kaposi. It was noted tachycardia in 15 cases, decrease of heart sounds in 12, arterial hypotension in seven, systolic murmur in 8, galop rhythm in 7, pericardial friction rub in 3, arrhythmia in 2. Four patients had congestive heart failure. The EKG showed sinus tachycardia in 18, ST and T changes in 10, low voltage in 5, ST segment elevation in 5 and extrasystoles in 3 cases. The echocardiogram findings were: pericardial effusion in 9 cases and 9 had ventricular dysfunction. CONCLUSION: The cardiac lesions were very important even in patients without clinical signals. We need others prospective studies with viral identification trying to detect specific lesions of HIV.

[Severe tricuspid valve insufficiency due to papillary anterior muscle infarction of the right ventricle secondary to neonatal hypoxia]. / Insuficiência tricúspide severa por infarto do músculo papilar anterior do ventrículo direito secundário à hipóxia neonatal.

A fatal case of a stressed newborn baby who developed tricuspid insufficiency due to an anterior papillary muscle infarction of the right ventricle, related to perinatal anoxia is reported. The baby needed resuscitation management and had a systolic murmur soon after birth. The echocardiographic examination showed a flail anterior leaflet of the tricuspid valve due to an avulsed anterior papillary muscle. There was rupture of chordae tendineae between anterior and posterior leaflet. Necrosis and calcium deposit were found in the area. The conclusion was that the anoxia in perinatal period, in most cases, causes transient lesions, but in few cases might be severe and fatal. It is important a complete cardiac evaluation in every case of prolonged perinatal stress.