Reversal of the anticoagulant and anti-hemostatic effect of low molecular weight heparin by direct prothrombin activation

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

Effect of passive pneumoperitoneum on oesophageal pressure, cardiovascular parameters and blood gas analysis in horses

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Neonatal meningitis according to the microbiological diagnosis: a decade of experience in a tertiary center

The aim of this study was to evaluate the incidence of and mortality due to meningitis and compare data according to microbiological diagnosis. This was a ten-year retrospective study conducted at a neonatal intensive care unit (NICU). Newborns with meningitis confirmed by positive CSF culture were included; those with congenital infection or malformations that made lumbar puncture impossible were excluded. The variables investigated were birth weight, gestational and postnatal age, procedures, hematological and CSF parameters, and complications. Parametric and non-parametric tests were used (statistical value p<0.05). The incidence of meningitis was 0.6% and mortality was 27%. of the 22 cases, 59% involved Gram-negative bacteria; 36% Gram-positive and 5% fungi. The groups did not differ in relation to birth weight, gestational and postnatal age, procedures or hematological and CSF parameters. Sepsis, convulsions and deaths were frequent in both groups, without statistical difference. Gram-negative cases showed abscesses and higher frequency of ventriculitis and hydrocephaly. Meningitis was infrequent, but presented high mortality and frequent complications.

Alterations in pulmonary structure by elastase administration in a model of emphysema in mice is associated with functional disturbances

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

PLANT-TO-SEED TRANSMISSION of CURTOBACTERIUM FLACCUMFACIENS pv. FLACCUMACIENS IN A DRY BEAN CULTIVAR

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

Molecular and parasitological survey of Hepatozoon canis (Apicomplexa : Hepatozoidae) in dogs from rural area of São Paulo state, Brazil

Hepatozoon canis is a protozoan that infects dogs and is transmitted by the ingestion of the brown dog tick, Rhipicephalus sanguineus. Two distinct species of Hepatozoon genus can infect dogs, H. canis and H. americanum. Routine tests to detect the disease are based on direct examination of gametocytes on Giemsa-stained blood smears. The objectives of this study were the investigation of infection prevalence in rural area dogs, the comparison of diagnostics by blood smear examination and polymerase chain reaction (PCR), and the association of infection with tick infestation. Blood smears, collected by puncture of the cephalic vein and ear margin capillary bed from 150 dogs, were examined. This technique detected 17 positive animals (11.3%), with 14 (9.3%) in peripheral blood and seven (4.7%) in cephalic vein blood. PCR tests detected 80 (53.3%) positive animals. R. sanguineus and Amblyomma spp. were found in 36 of the dogs (24%), in equal proportions. The identified species for Amblyomma genus were A. cajennense and A. ovale. Data analysis showed that PCR was much more sensitive when compared to blood smear examination. Hepatozoon species was previously identified as closely related to H. canis.

Propolis Standardized Extract (EPP-AF (R)), an Innovative Chemically and Biologically Reproducible Pharmaceutical Compound for Treating Wounds

The aim of this study was to develop a formulation, containing the propolis standardized extract (EPP-AF (R)), which can assist in the healing of skin lesions. To achieve this objective the antimicrobial activity and chemical composition of the propolis extract was determined. The final product was subjected to in vitro and in vivo pre-clinical evaluation. The broth macrodi-lution method was used to determine the antimicrobial activity of the extracts and formulations against the microorganisms most commonly found in burns, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. Wistar rats with puncture wounded skin were used to evaluate the wound healing properties of propolis. The results of chemical and biological characterization demonstrated the batch-to-batch reproducibility of the standardized extract which is an unprecedented result. The antimicrobial and wound healing activity of the pharmaceutical studied showed the best results when samples contain 3.6% propolis, suggesting that this is the most promising composition.

Anemia em crianças menores de dois anos atendidas nas unidades básicas de saúde no Estado de São Paulo, Brasil

The present study sought to determine the prevalence of anemia in 2,992 children, aged between 6 and 23 months, who voluntarily attended 160 Basis Health Care Units, located in 63 cities of the 5 Regional Health Coordinating areas of the State of S. Paulo, Brazil. Blood samples were collected by venous puncture and hemoglobin was measured by the cianometahemoglobin method. The WHO criteria for the diagnosis of anemia (Hb < 11.0 g/dl) were used: 59.1% of the children were shown to be anemic, with prevalence varying from 47.8% to 68.7% in the 5 RHCs. RHC 1, which comprises the Greater S. Paulo Region, presented a prevalence of anemia significantly lower than the other 4 RHCs, which cover the rest of the State. Hemoglobin levels 9.5 g/dl were found in 25.1% of the children. Anemia was more frequent in male children in male children, in those born, with a weight of less than 3,000 gr, in those who were breastfed for less than 2 months and in those that who presented some degree of energy deficient proteic malnutrition, according to Gomez's criteria. This is the first of a series of 4 articles whose purpose is the determining the prevalence of anemia in the State of S. Paulo and of testing the intervention alternatives with a view to curtailing the incidence of this pathology which today is the most prevalent nutritional disturbance in the world.

Association between polyclonal B cell activation and the presence of autoantibodies in mice infected with Yersinia enterocolitica O:3

Eight-week old conventional female Swiss mice were inoculated intravenously with Yersinia enterocolitica O:3. A second group of normal mice was used as control. Five mice from each group were bled by heart puncture and their spleens were removed for spleen cell collection on the 3rd, 5th, 7th, 10th, 14th and 21st day after infection. Immunoglobulin-secreting spleen cells were detected by the isotype-specific protein A plaque assay. Total immunoglobulin levels were determined in mouse serum by single radial immunodiffusion and the presence of autoantibodies was determined by ELISA. We observed a marked increase in the total number of cells secreting immunoglobulins of all isotypes as early as on the 3rd day post-infection and the peak of secretion occurred on the 7th day. At the peak of the immunoglobulin response, the total number of secreting cells was 19 times higher than that of control mice and most immunoglobulin-secreting cells were of the IgG2a isotype. On the 10th day post-infection, total serum immunoglobul in values were 2 times higher in infected animals when compared to the control group, and continued at this level up to the 21st day post-infection. Serum absorption with viable Y. enterocolitica cells had little effect on antibody levels detected by single radial immunodiffusion. Analysis of serum autoantibody levels revealed that Y. enterocolitica infection induced an increase of anti-myosin and anti-myelin immunoglobulins. The sera did not react with collagen. The present study demonstrates that Y. enterocolitica O:3 infection induces polyclonal activation of murine B cells which is correlated with the activation of some autoreactive lymphocyte clones.

Use of fibrin glue derived from snake venom in testicular biopsy of rams

Sequelae due to testicular biopsy such as hemorrhage, adhesion and fibrosis may be limiting factors to the use of this surgical procedure. Fibrin glue (FG) derived from snake venom was used to minimize these sequelae, as well as to evaluate its healing property in tunica vaginalis and scrotal skin of rams. Applicability of fibrin glue derived from snake venom was tested in different tissues of other animals such as in sciatic nerve and colon of rats and skin of rabbits. In the present study, 30 healthy adult rams were used. They were divided into 3 groups of 10 animals each as follows: G1: fibrin glue group (application of fibrin glue on puncture sites and skin incisions after bilateral testicular biopsy with a Tru-Cut needle); G2: swab/nylon group (hemostasis by compression with a swab on puncture sites and skin suturing with nylon after biopsy) and G3: control group (the animals were not subjected either to biopsy or to surgery). On the 20th day after biopsy, the presence of adhesion strands between the sites of skin incision and testicle was evaluated by palpation Adhesion strands were found in three testicles (15%) in G1 and in two testicles (10%) in G2. One hundred days after biopsy, orchiectomy was carried out and the material collected was assessed for subcutaneous (SC) and/or tunica vaginalis adhesions. G3 did not present any abnormality. Groups G1 and G2 presented four testicles each (20%) with adhesion between the tunics at biopsy site. On the other hand, subcutaneous adhesions were found once (5%) in G1 and three times (15%) in G2. Fibrin glue showed to be of easy application, required short postoperative monitoring, presented fast and good-quality healing property and tended to reduce formation of subcutaneous adhesion.

Peridural com ropivacaina a 1%: Experiencia com volume proporcional a estatura

Background and Objectives - Ropivacaine - a local amino amide anesthetic agent - is a plain S enantiomer which makes it a potent and low toxicity drug. The aim of our study was to evaluate 1% ropivacaine for epidural block in lower doses than those described in the literature. Methods - Thirty-eight patients, physical status ASA I and II, aged 15 to 70 years, weighing 50 to 100 kg were selected. Premedication consisted of 15 mg oral midazolam given 60 min before anesthesia induction. In the OR, after standard monitoring a catheter was inserted intravenously to administer 10 ml.kg-1 Ringers lactate solution. Epidural puncture was performed with the patient in the sitting position and 1% ropivacaine was administered in a volume corresponding to 10% of patient's height in centimeters. With the patient in the supine position, motor blockade intensity, temperature sensitivity and sensory block extension at 1, 3, 5, 7, 10, 15, 20, 30 minutes after drug injection were evaluated. Blood pressure, heart rate and adverse side effects during the course of anesthesia and in the post-anesthetic period were also observed. In the recovery room patients were followed-up until motor blockade intensity temperature sensitivity and sensory block had returned to level L2. Results - Mean values were 41.4 years of age, 68.8 kg of body weight and 165 cm height. Upper thermal blockade level was T4 and upper sensory block level was T6. Most patients showed motor block level 1 (Bromage scale) after 30 minutes of observation. Motor block mean duration was 254 minutes and temperature sensitivity 426 minutes. Only three patients had complications: two cases of hypotension and one of bradycardia. Conclusions - In the volumes used in this study, ropivacaine produced adequate analgesia and a less intense lower limb motor block which, however, was sufficient to allow for surgical procedures with low incidence of side-effects.

Effects of G a3 dosis associated with organosilicone on sweet oranges in Brazil

GA3 was tested in sweet oranges 'Pera' and 'Hamlin' for delay the picking time without loosening of fruit quality for processing. Hamlin is the firths cv processed in Brazil and Pera is a mid season cv and extending their period of processing is important. Two experiments were made at the Citrus Experimental Station during 1996 season. The treatments are 5 ppm of GA 3 + 0,05% Silwett L-77® (organosilicone), 10 ppm of GA 3 + 0,05% of Silwet L-77®, 20 ppm of GA3 + 0,1% Herbitensil® (Noniphenoloxietilate 40%m/v + isopropilic alcohol 15% m/v) and control, repeated 7 times for Hamlin and 8 times for Pera, with one tree each parcel. The treatments were applied in May 1996, at the stage of greenish yellow colour of the fruits. Evaluations were made each 20 days interval till the final picking, It was analysed fruit quality and retention force for picking and puncture resistance. The results showed no differences for fruit quality of Hamlin from July to mid September and for Pera till September. After some differences occurred. The GA3 treatments were effectives in maintain the fruit retention force for both cvs for 120 days after application. In relation to fruit puncture resistance the treatments with GA3 differed of the control for both cvs, accordingly with the doses and mixtures. The colour index was better maintained with 5ppm of GA3 plus 0,05% of Silwet L-77®. The total fruit production did not differ for both cultivars.

Efeitos de diferentes doses de tenoxicam sobre a medula espinhal e as meninges. Estudo experimental em caes

Background and Objectives. The analgesic actions of nonsteroidal anti-inflammatory drugs (NSAID) result from the inhibition of the peripheral synthesis of prostaglandins. In spite of the emphasis on the peripheral action, several studies have shown the potential central action of such drugs. In rats, NSAID doses insufficient to block pain when systemically administered were effective when intrathecally injected. This effect could be mediated by interaction with descending serotoninergic ways together with neurotransmission modulation of glycine or N-methyl-D-aspartate receptors. Our goal was to study the effect of different tenoxican doses in the histology of dogs spinal cord and meninges. Methods. Thirty two dogs (7 to 17 kg) were randomly distributed in four groups: G1 - Control with distilled water (DW); G2 - 2 mg tenoxican diluted in DW; G3 - 4 mg tenoxican diluted in DW; G4 - 10 mg tenoxican diluted in DW in a constant volume of 1 ml. Anesthesia was induced with etomidate and fentanyl and dural puncture was performed with a 25G spinal needle in interspace L6-7. Animals were observed for 72 hours and subsequently euthanized by electrocution. Lumbar and sacral spinal cord segments were removed for further histologic examination. Results. All animals were clinically normal during the observation period and there has been no histologic alteration of the nervous system and meninges. Conclusions. In our experimental model intrathecal tenoxican doses up to 10 mg have not triggered nervous tissue or meningeal injuries in dogs.

Anesthesia of fish with benzocaine does not interfere with comet assay results

Fish blood erythrocytes are frequently used as sentinels in biomonitoring studies. Usually, fish blood is collected by painful cardiac or caudal vein punctures. Previous anesthesia could decrease animal suffering but it is not known at present whether anesthesia can cause confounding effects. Therefore, using the alkaline single cell gel (SCG)/comet assay with blood erythrocytes of the cichlid fish Nile tilapia, we tested for a possible modulation of induced DNA damage (methyl methanesulfonate; MMS) by the anesthetic benzocaine administered by bath exposure (80mg/l for ∼10min). Furthermore, benzocaine (80-600mg/l) was tested for its genotoxic potential on fish erythrocytes in vitro and for potential interactions with two known genotoxins (MMS and hydrogen peroxide). Our results did neither indicate a significant increase in the amount of DNA damage (even after a 48h follow-up), nor indicated interactions with MMS-induced DNA damage when fish were exposed to benzocaine in vivo. There was also no increase in DNA damage after in vitro exposure of fish erythrocytes to benzocaine. Clear concentration-related effects were observed for the two genotoxins in vitro, which were not significantly altered by the presence of benzocaine. These results suggest that anesthesia of fish does not confound comet assay results and the use of blood samples from anesthetized fish can be recommended with regard to animal welfare. © 2002 Elsevier Science B.V. All rights reserved.

Standardization of platelet parameters determination in blood samples collected with EDTA

The Abbott Cell-Dyn 3000 automated hematological analyzer prepares a histogram of platelet volume, which is measured by a technique using electrical impedance, inferring from its platelet count (PLT), mean platelet volume (MPV) and platelet distribution width (PDW). This equipment also calculates the plateletcrit (PCT). These platelet parameters may be important to evaluate platelet function, but they require standardization, because platelets swell when in contact with ethylenediaminetetra-acetic acid salts, hence an increase in blood sample storage time produces artificially increased results. To assess the effect of storage time on MPV, PLT, PDW and PCT, blood samples from 23 sickle cell anemia patients during steady state (Group I) and 50 from healthy controls (Group II) were placed in Vacutainer® tubes with dipotassium ethylenediaminetetra-acetic acid and measured over a period of 1440 minutes (24 h) at the following times: immediately after the venipuncture (time 0), 15, 30, 60, 120, 240, 360, 480 and 1440 minutes. The mean values of MPV and PCT were significantly increased (p<0.00001) along the storage time in both groups. The mean values of PLT and PDW were practically stable (p>0.05) throughout the storage time in both groups.