DIAGNOSTIC REFERENCE LEVELS BASED ON PATIENT BODY MASS INDEX FOR SELECT INTERVENTIONAL PROCEDURES IN MINAS GERAIS/BRAZIL.

This work establishes local diagnostic reference levels (DRLs) in interventional radiology based on adult patient body mass index (BMI). The monitoring was carried out from 23 institutions and patient data from 3015 procedures were collected, being 907 Catheterism (CAT), 921 Percutaneous transluminal coronary angioplasty (PTCA) and 1187 CAT/PTCA and 6 BMI ranges were taken, going from under 18 up to 40 kg·m-2. It is presented that 18 initial DRL values to be used in the county of Minas Gerais. The overall 75th kerma-area product (KAP), commonly considered DRL, in Gy·cm2, is 94.6(SD 119)-CAT, 88.6(SD 121)-PTCA and 33.0(SD 47.6)-CAT/PTCA. In considering KAP-BMI individual values, one obtain the min-max ranges, in Gy·cm2, 3.2-BMI A to 101-BMI B for CAT, 65-BMI A to 102-BMI F for PTCA and 10.4-BMI A to 59.2-BMI E for CAT/PTCA. The KAP-BMI approach has shown to be feasible as a DRL optimization process.

In vitro cytotoxic effects of 5-Fluorouracil on isolated murine ovarian preantral follicles.

5 fluorouracil (5FU), an antineoplastic drug, is often utilized in the therapeutic regimen for several types of cancer, including the hepatoblastoma in children. The effects of 5FU on the population of ovarian preantral follicles, which is the largest oocyte reservoir, is still poorly understood. The integrity of the ovarian preantral follicle pool is important for lifelong fertility. The better understanding of such effects may favor intervention strategies to protect fertility in 5FU-treated children and women coping with cancer. To analyze the effects of 5FU on isolated murine secondary follicles in vitro, ovaries were collected from young mice (28-30 days old), and secondary follicles were isolated and cultured for 12 days in basic culture medium, with or without 5FU at concentrations of 0.3 mM, 1 mM, 3 mM, 10 mM, and 30 mM. In the in vitro study, we analyzed the percentage of morphologically normal follicles, antrum formation, follicular diameter, and hormone production. On day 12, oocytes were recovered for in vitro maturation. 5FU treatment did not alter the percentage of morphologically normal follicles. On day 12, only 1, 10, and 30 mM 5FU significantly reduced the percentage of antrum. From day 4 onwards, 5FU treatments significantly reduced follicle diameter. The meiosis resumption rate was significantly lower in all 5FU treatments. 5FU concentrations ≥3 mM reduced estradiol levels. In conclusion, 5FU does not affect follicular morphology. However, 5FU deleteriously affects follicular growth, estradiol production, and oocyte maturation in isolated ovarian follicles.

Cilostamide affects in a concentration and exposure time-dependent manner the viability and the kinetics of in vitro maturation of caprine and bovine oocytes.

This study investigated: 1) the kinetics of oocyte chromatin configuration during in vitro maturation (IVM) of caprine and bovine oocytes; and 2) the effect of in vitro pre-maturation (IVPM) with cilostamide with or without association of the follicular wall (FW) on the same parameters. In experiment I, cumulus-oocyte complexes (COCs) were cultured in vitro in a standard maturation medium for 6, 12, 18 or 30 h. For experiment II, the COCs were cultured for 30 h, either in a standard IVM medium or in IVPM containing cilostamide (10 or 20 µM) and FW alone or in combination, for 6 or 12 h before the onset of maturation. The MII rate was similar (P > .05) between 18 and 30 h of maturation, both of which were higher (P < .05) than 6 and 12 h IVM in both species (Experiment I). Contrary to caprine, all IVPM treatments presented a higher (P < .05) percentage of bovine oocytes arrested at the GV stage than the control treatment after 6 h of culture. The percentage of MII oocytes after 30 h (IVPM+IVM) of culture in bovine oocytes treated with 10 µM cilostamide associated with FW and FW alone cultured for 6 h presented MII percentages similar to the control. However, in caprine, these treatments significantly reduced the percentages of MII in relation to the control treatment (Experiment II). In conclusion, the combination of concentration-exposure time to cilostamide during IVPM delayed meiotic progression in bovine after 6 and 12 h of culture. However, overall the culture period (IVPM+IVM) influenced the oocyte chromatin configuration and kinetics in both species.

Dose-dependent effects of frutalin on in vitro maturation and fertilization of pig oocytes.

The aim of the present study was to evaluate the effect of frutalin (FTL) on in vitro maturation (IVM), and fertilization (IVF) of pig oocytes. In the Experiment 1, cumulus-oocyte complexes (COCs) were submitted to IVM in maturation medium alone or supplemented with different FTL concentration (0.6, 6 and 60 µg/mL), or 0.3 µg/mL doxorubicin (DXR). After IVM, some oocytes were evaluated for chromatin configuration, and the remaining oocytes were submitted to in vitro fertilization. In Experiment 2, matured oocytes were fertilized in IVF medium alone (control) or in presence of different FTL concentration (0.6, 6 and 60 µg/mL), or 0.3 µg/mL DXR. After 18 h post fertilization, the endpoints penetration rate, monospermy, spermatozoa per oocyte, and the IVF efficiency were evaluated in both experiments. In Experiment 1, 6 and 60 µg/mL FTL, as well as DXR increased (P < 0.05) the rate of oocytes with abnormal chromatin configuration when compared to oocyte matured in control medium alone or supplemented with 0.6 µg/mL FTL. The percentage of meiotic resumption in oocytes cultured with 60 µg/mL FTL or DXR was less (P < 0.05) than in the other treatments. Moreover, oocytes matured with 6 or 60 µg/mL FTL and DXR had a lesser IVM efficiency when compared to those matured with 0.6 µg/mL FTL or in control medium. Additionally, there was a greater (P < 0.05) with culture in a medium containing 6 µg/mL FTL for the rate of partenogenetically activated oocytes when compared with the other treatments. Culturing of COCs during IVM in a medium containing 6 or 60 FTL resulted in a lesser (P < 0.05) sperm penetration and spermatozoa/oocyte rates when compared to other treatments, and IVF efficiency was less (P < 0.05) than that in control medium alone or with a medium containing 0.6 µg/mL FTL. In Experiment 2, culturing in a medium containing 0.6 µg/mL FTL resulted in greater (P < 0.05) monospermy and IVF efficiency rates when compared to culturing in the control medium. In addition, culturing in a medium with 6 and 60 µg/mL FTL resulted in a lesser (P < 0.05) spermatozoa penetration, sperm/oocyte rates and IVF efficiency, although there were greater (P < 0.05) monospermy rates. In conclusion, culturing in a medium containing 0.6 µg/mL FTL resulted in lesser spermatozoa penetration rates and number of spermatozoa/oocyte increasing the IVF efficiency without harmful effects. Use of a greater concentration of FTL in the medium has toxic effects during oocyte maturation and results in a reduced IVF efficiency.

Relationship between follicular dynamics and oocyte maturation during in vitro culture as a non-invasive sign of caprine oocyte meiotic competence.

The search for non-invasive signs of oocyte meiotic competence is very important for the development of in vitro follicle culture (IVFC) systems. The aims of the present study were: (1) to investigate the effect of in vitro maturation (IVM) of in vivo grown goat COCs, in group or individually, on oocyte chromatin configuration (Experiment 1), and (2) the influence of IVFC period (12 vs. 18 days) on the ability of the oocyte to resume meiosis immediately after IVFC (before in vitro maturation; IVM), or after IVM (Experiment 2). In experiment 1, in vivo grown cumulus-oocyte complexes (COCs) were submitted to IVM in groups (10 COCs/100 µL-drop) or individually (1 COC/10 µL-drop), and chromatin configuration was assessed. In experiment 2, isolated follicles were individually cultured for 12 or 18 days, and submitted to individual IVM afterwards. The following end points were evaluated: follicular growth and morphology, oocyte diameter, viability and chromatin configuration, as well as individual follicular estradiol production. Similar maturation rates were obtained between in vivo grown COCs matured individually and in groups (66.7% vs. 63.6%, respectively) (Experiment 1). Only after 18 days of IVFC, oocytes were able to grow during IVM, reaching a mean oocyte diameter of 119 µm. Also, this treatment produced the highest rate of metaphase II oocytes (46.2% out of the total number of cultured follicles). Finally, it was observed that follicles with a daily growth rate >7.1 µm/day (fast-growing) and that reached at least 600 µm in diameter, were more likely (P < 0.05) to produce oocytes capable of attaining MII. In conclusion, caprine oocytes can be individually matured in vitro, as efficiently as in groups. This result was essential to pair in vitro follicle development and in vitro oocyte maturation with specific individual follicles. Using this approach, it was possible to establish non-invasive signs for the efficiency of IVFC based on follicle daily growth rate and diameter, and oocyte diameter: follicle daily growth >7 µm, follicle diameter of at least 600 µm, and oocyte diameter ≥120 µm. In addition, 18 days seems to be the most suitable culture time for caprine early antral follicles.

In vitro growth and maturation of isolated caprine preantral follicles: Influence of insulin and FSH concentration, culture dish, coculture, and oocyte size on meiotic resumption.

The aims of this study were: (1) to evaluate the effect of different insulin concentrations, alone or in combination with either a fixed FSH concentration or increasing FSH concentrations on the in vitro culture of isolated caprine preantral follicles and (2) to analyze the efficiency of two IVM media and maturation culture systems (with or without coculture with in vivo grown oocytes) on the meiosis resumption. Secondary follicles were cultured for 18 days in a basic medium supplemented with low- or high-insulin concentration alone or with a fixed FSH concentration or with increasing FSH concentrations. Oocytes grown in vivo or in vitro were matured alone or cocultured. The high-insulin concentration associated with fixed FSH treatment had higher meiotic resumption rate (P < 0.05) and was the only treatment capable of producing oocytes in metaphase II. The rates of germinal vesicle, germinal vesicle breakdown, metaphase I, metaphase II (MII), meiotic resumption, and oocyte diameter were similar between the maturation media. In conclusion, a basic medium supplemented with 10-µg/mL insulin and 100-µg/mL FSH throughout the culture period improved meiotic resumption rate and produced MII oocytes from caprine preantral follicles cultured in vitro. The MII rate was similar between in vivo and in vitro grown oocytes ≥110 µm.

Caprine ovarian follicle requirements differ between preantral and early antral stages after IVC in medium supplemented with GH and VEGF alone or in combination.

The aim of the present study was to evaluate the effect of growth hormone (GH) and vascular endothelial growth factor (VEGF) added alone, sequentially or in combination, in the presence of insulin at physiological concentration (10 ng/mL) on the IVC of two different follicular categories: preantral (experiment 1; Exp.1) and early antral (experiment 2; Exp.2). Isolated follicles were individually cultured for 24 (Exp.1) and 18 days (Exp.2) in the following treatments: αMEM+ (Control), or Control medium supplemented with 50 ng/mL GH (GH), 100 ng/mL VEGF (VEGF), the combination of both (GH + VEGF), GH during the first 12 days and VEGF from Day 12 until the end of the culture (GH/VEGF) and vice versa (VEGF/GH). At the end of the culture, cumulus-oocyte complexes from in vitro-grown follicles were recovered and subjected to IVM. The following end points were evaluated: Follicle morphology, growth rates and antrum formation, production of estradiol, progesterone and testosterone, oocyte viability and meiotic stage, as well as relative expression of LHR, Amh, HAS2, PTGS2, CYP17, CYP19A1, and 3ßHSD. A considerable amount of viable fully grown oocytes were recovered after the IVC of early antral follicles in all treatments. Nevertheless, the GH treatment presented the highest percentage of fully grown oocytes (60%), mean oocyte diameter (117.74 ± 2.61 µm), and meiotic resumption (50%). Furthermore, GH treatment produced higher (P < 0.05) rates of metaphase II oocytes than all the other treatments, and similar LHR, Amh, and PTGS2 transcript levels to in vivo. Contrary to early antral follicles, preantral follicles were not affected by medium supplementation. In conclusion, the addition of GH to a culture medium containing physiological concentrations of insulin improves oocyte growth and maturation after the IVC of goat early antral follicles.

Effect of heat stress on the survival and development of in vitro cultured bovine preantral follicles and on in vitro maturation of cumulus-oocyte complex.

The deleterious effect of heat stress (HS) on competence of oocytes from antral follicles is well recognized, but there is a lack of data regarding its impact on the viability and growth of preantral follicles. In this study, we used in vitro preantral follicle cultures to investigate the effects of HS on the following parameters: survival and development of primordial follicles after in vitro culture of ovarian fragments (experiment I); growth and antrum formation of isolated advanced secondary follicles (experiment II); and maturation rates after in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) from antral follicles (>2-6 mm) grown in vivo (experiment III). Furthermore, the following end points were evaluated in all experiments: follicle/oocyte survival, reactive oxygen species (ROS), estradiol (E2) and progesterone (P4) production, as well as mRNA expression for select genes related to stress (HSP70) and apoptosis (MCL1 and BAX). In all experiments, HS consisted of exposing the structures (ovarian fragments, isolated preantral follicles and COCs) to 41 °C for 12 hours and then to 38.5 °C until the end of the culture (7 days for experiments I and II and 24 hours for experiment III). The temperature for the control group was held at 38.5 °C for the entire culture period. Heat stress increased (P < 0.05) the percentage of developing follicles (intermediate, primary, and secondary follicles) at 12 hours and increased levels of ROS at all evaluated time points (12, 24 hours, and D7), when compared to the control (experiment I). Heat stress did not affect (P > 0.05) any identified end points when preantral follicles were cultured in their isolated form (experiment II). However, in experiment III, HS decreased (P < 0.05) both the rates of metaphase II after 24 hours and E2 production at 12 hours of IVM. Moreover, HS increased (P < 0.0001) levels of P4 after IVM and ROS production at every evaluated time point, compared with the control (12 and 24 hours). In conclusion, HS caused: (1) early activation of primordial follicles; (2) an increase in ROS production by early preantral follicles enclosed in ovarian tissue and by COCs; (3) a short-term reduction of E2 production by COCs; and (4) an increase in P4 secretion from COCs. However, HS did not affect in vitro culture of advanced isolated secondary follicles. Experimental evidence indicates that preantral follicles are less sensitive to HS than COC.

Survival capacity of Mycoplasma agalactiae and Mycoplasma mycoides subsp capri in the diluted semen of goat bucks and their effects on sperm quality.

This study examines the viability of Mycoplasma agalactiae (Ma) and Mycoplasma mycoides subsp capri (Mmc) during 150 minutes of incubation at 37 °C in contaminated diluted semen (DS) doses. The effects of the presence of both microorganisms on sperm viability, motility, and morphology were also examined. In a second experiment, the viability of Ma and its effects on sperm viability were determined in ejaculate samples and skimmed milk semen extender samples. Ma and Mmc were able to survive in DS at concentrations considered infectious, and no significant differences in mean concentrations were detected (7.1 log colony-forming units [CFU]/mL). However, initial concentration of Ma declined (P < 0.05) from 7.5 to 6.9 log CFU/mL and Mmc declined (P < 0.05) from 7.7 to 7.1 log CFU/mL after incubation. Conversely, ejaculate concentrations of Ma increased significantly (from 7.1 to 7.4 log CFU/mL, P < 0.05). These observations suggest that the natural breeding medium is more suitable for Ma than the medium used for artificial insemination (AI). The presence of Mmc slightly reduced sperm viability in the DS (from 21.7% to 16.6%, P < 0.05). The absence of major effects on sperm quality could lead to the unnoticed use of semen contaminated with Ma and Mmc for AI. As both bacteria were able to survive the conditions of ejaculates and semen doses, these findings suggest a risk of venereal transmission of contagious agalactia and support the use of mycoplasma-free semen samples for (AI).

Equine spermatozoa stored in the epididymis for up to 96h at 4°C can be successfully cryopreserved and maintain their fertilization capacity.

After injury or death of a valuable male, recovery of epididymal spermatozoa may be the last chance to ensure preservation of its genetic material. The objective of this research was to study the effect of sperm storage, at 4°C up to 96h, in the epididymides obtained from castrated horses and its effect on different functional sperm parameters. Aims were to study the effect of (1) sperm storage on viability and chromatin condensation; (2) pre-incubation of recovered epididymal sperm in the freezing extender, prior cryopreservation, on viability and chromatin condensation; and (3) freezing-thawing on viability, chromatin condensation, ROS generation, protein tyrosine phosphorylation and heterologous fertilization rate (ICSI and IVF using bovine oocytes) of sperm recovered from the epididymis up to 96h post castration. The average volume (720±159µL) and the concentration (6.5±0.4×10(9) spermatozoa/mL) of sperm recovered from the epididymis were not affected by storage. Sperm viability after refrigeration at 4°C for up to72h was similar (P<0.01). The effect of sperm dilution in the freezing media showed similar values up to 48h, while viability was preserved up to 72h (P<0.01). Cryopreserved spermatozoa show similar viability between different storage times. Chromatin condensation was not affected by storage time; however, incubation for 30min in freezing medium and freezing-thawing process induced an increase in the chromatin decondensation. ROS generation was not affected by storage up to 96h. Epididymal storage did not affect sperm protein tyrosine phosphorylation patterns; although the pattern of phosphorylation changed to strong staining of the equatorial segment when the sperm where capacitated in sperm-TALP. Finally, successful and similar pronuclear formation (analyzed by ICSI) and in vitro penetration (evaluated with bovine zone free oocyte) was observed using cryopreserved sperm obtained from prolong epididymal storage at 4°C. In conclusion, cryopreservation of epididymal stallion sperm stored for up to 72h in the epididymis at 4°C, maintain both viability and ability to fertilize in vitro.

The alga Bryothamnion seaforthii contains carbohydrates with antinociceptive activity.

Bryothamnion seaforthii, a red alga common to the Northeastern coast of Brazil, was used to prepare the protein fraction F0/60 by ammonium sulfate precipitation. The chromatography of F0/60 on DEAE-Sephadel column resulted in two lectin fractions, PI and PII, which have antinociceptive properties in rodents. We determined the antinociceptive activity of the PII fraction and of a carbohydrate-containing fraction (CF) in mice. The CF was prepared from the dried algae, after digestion with 100 mM sodium acetate, pH 6.0, containing 5 mM cysteine, EDTA and 0.4% papain, at 60 masculine C. A 10% cetylpyridinium chloride was added to the filtrate, and the precipitate was dissolved with 2 M NaCl:ethanol (100:15, v/v) followed by the carbohydrate precipitation with ethanol. The final precipitate, in acetone, was dried at 25 masculine C. The PII fraction markedly inhibited acetic acid-induced abdominal writhing after ip administration (control: 27.1 +/- 2.20; PII 0.1 mg/kg: 5.5 +/- 1.85; 1 mg/kg: 1.6 +/- 0.72 writhes/20 min) and after oral administration (control: 32.0 +/- 3.32; PII 0.1 mg/kg: 13.1 +/- 2.50; 1 mg/kg: 9.4 +/- 3.96 writhes/20 min). PII was also effective against both phases of pain induced by 1% formalin (control, ip: 48.2 +/- 2.40 and 27.7 +/- 2.56 s; PII: 1 mg/kg, ip: 34.3 +/- 5.13 and 5.6 +/- 2.14 s; control, po: 44.5 +/- 3.52 and 25.6 +/- 2.39 s; PII 5 mg/kg, po: 26.5 +/- 4.67 and 15.3 +/- 3.54 s for the 1st and 2nd phases, respectively) and in the hot-plate test. The CF (ip) also displayed significant antinociceptive properties in all tests but at higher doses (1 and 5 mg/kg, ip and po). Thus, CF at the dose of 5 mg/kg significantly inhibited writhes (ip: 7.1 +/- 2.47 and po: 14.5 +/- 2.40 writhes/20 min) as well as the 1st (po: 19.6 +/- 1.74 s) and 2nd (po: 7.1 +/- 2.24 s) phases of the formalin test compared to controls ip and po. The antinociceptive effects of both the PII and CF in the formalin and hot-plate tests were prevented at least partially by pretreatment with the opioid receptor antagonist naloxone (2 mg/kg, sc). Moreover, both fractions retained antinociceptive activity in the acetic acid-induced writhing test following heating, a procedure which abolished the hemagglutinating activity of the fraction, presumably due to lectins also present. Finally, both fractions also prolonged the barbiturate-induced sleeping time. These results indicate that carbohydrate molecules present in the PII (26.8% carbohydrate) and CF (21% of the alga dried weight) obtained from B. seaforthii display pronounced antinociceptive activity which is resistant to heat denaturation and is mediated by an opioid mechanism, as indicated by naloxone inhibition.

The alga Bryothamnion seaforthii contains carbohydrates with antinociceptive activity

Bryothamnion seaforthii, a red alga common to the Northeastern coast of Brazil, was used to prepare the protein fraction F0/60 by ammonium sulfate precipitation. The chromatography of F0/60 on DEAE-Sephadel column resulted in two lectin fractions, PI and PII, which have antinociceptive properties in rodents. We determined the antinociceptive activity of the PII fraction and of a carbohydrate-containing fraction (CF) in mice. The CF was prepared from the dried algae, after digestion with 100 mM sodium acetate, pH 6.0, containing 5 mM cysteine, EDTA and 0.4 percent papain, at 60ºC. A 10 percent cetylpyridinium chloride was added to the filtrate, and the precipitate was dissolved with 2 M NaCl:ethanol (100:15, v/v) followed by the carbohydrate precipitation with ethanol. The final precipitate, in acetone, was dried at 25ºC. The PII fraction markedly inhibited acetic acid-induced abdominal writhing after ip administration (control: 27.1 ± 2.20; PII 0.1 mg/kg: 5.5 ± 1.85; 1 mg/kg: 1.6 ± 0.72 writhes/20 min) and after oral administration (control: 32.0 ± 3.32; PII 0.1 mg/kg: 13.1 ± 2.50; 1 mg/kg: 9.4 ± 3.96 writhes/20 min). PII was also effective against both phases of pain induced by 1 percent formalin (control, ip: 48.2 ± 2.40 and 27.7 ± 2.56 s; PII: 1 mg/kg, ip: 34.3 ± 5.13 and 5.6 ± 2.14 s; control, po: 44.5 ± 3.52 and 25.6 ± 2.39 s; PII 5 mg/kg, po: 26.5 ± 4.67 and 15.3 ± 3.54 s for the 1st and 2nd phases, respectively) and in the hot-plate test. The CF (ip) also displayed significant antinociceptive properties in all tests but at higher doses (1 and 5 mg/kg, ip and po). Thus, CF at the dose of 5 mg/kg significantly inhibited writhes (ip: 7.1 ± 2.47 and po: 14.5 ± 2.40 writhes/20 min) as well as the 1st (po: 19.6 ± 1.74 s) and 2nd (po: 7.1 ± 2.24 s) phases of the formalin test compared to controls ip and po. The antinociceptive effects of both the PII and CF in the formalin and hot-plate tests were prevented at least partially by pretreatment with the opioid receptor antagonist naloxone (2 mg/kg, sc). Moreover, both fractions retained antinociceptive activity in the acetic acid-induced writhing test following heating, a procedure which abolished the hemagglutinating activity of the fraction, presumably due to lectins also present. Finally, both fractions also prolonged the barbiturate-induced sleeping time...

Antinociceptive activity of sulfated carbohydrates from the red algae Bryothamnion seaforthii (Turner) Kütz. and B. triquetrum (S.G. Gmel.) M. Howe.

We report the antinociceptive activity, determined by the writhing, formalin and hot-plate tests in mice, of crude (F0/60), lectin and carbohydrate fractions isolated by ammonium sulfate precipitation (0 to 60%) from Bryothamnion seaforthii and B. triquetrum, species of red algae. Not only fraction F0/60 but also lectins from both species significantly inhibited acetic acid-induced abdominal contractions after intraperitoneal or oral administrations. In the formalin test, lectins (1 and 5 mg/kg, ip, and 5 to 20 mg/kg, po) inhibited the 1st and 2nd phases (5 and 20 min, respectively), but the effect occurred predominantly on the 2nd phase. The effects of the lectins were totally or partially reversed by naloxone (2 mg/kg, sc) in the 1st and 2nd phases, respectively. Experiments performed with lectins in the absence and presence of avidin (1 mg/kg, ip) and D-mannose (1 mg/kg, ip) showed that avidin did not interfere with the effect of B. seaforthii lectin but partially reversed the effect of B. triquetrum lectin. D-Mannose completely reversed the effects of both species. F0/60 fractions from both algae significantly increased the latency time in response to thermal stimuli, and naloxone reversed antinociception, indicating the involvement of the opioid system in both the peripheral and central effects of the fractions. In the writhing test, the carbohydrate fractions were the most active, inhibiting the contractions by 71 and 79% (B. triquetrum) and by 46 and 69% (B. seaforthii) at doses of 1 and 5 mg/kg, ip, respectively. Sulfated carbohydrate fractions of B. seaforthii and B. triquetrum, containing only about 5% protein as contaminants, are probably responsible for the antinociceptive effects of these red algae.

Antinociceptive activity of sulfated carbohydrates from the red algae Bryothamnion seaforthii (Turner) Kütz. and B. triquetrum (S.G. Gmel.) M. Howe

We report the antinociceptive activity, determined by the writhing, formalin and hot-plate tests in mice, of crude (F0/60), lectin and carbohydrate fractions isolated by ammonium sulfate precipitation (0 to 60 percent) from Bryothamnion seaforthii and B. triquetrum, species of red algae. Not only fraction F0/60 but also lectins from both species significantly inhibited acetic acid-induced abdominal contractions after intraperitoneal or oral administrations. In the formalin test, lectins (1 and 5 mg/kg, ip, and 5 to 20 mg/kg, po) inhibited the 1st and 2nd phases (5 and 20 min, respectively), but the effect occurred predominantly on the 2nd phase. The effects of the lectins were totally or partially reversed by naloxone (2 mg/kg, sc) in the 1st and 2nd phases, respectively. Experiments performed with lectins in the absence and presence of avidin (1 mg/kg, ip) and D-mannose (1 mg/kg, ip) showed that avidin did not interfere with the effect of B. seaforthii lectin but partially reversed the effect of B. triquetrum lectin. D-Mannose completely reversed the effects of both species. F0/60 fractions from both algae significantly increased the latency time in response to thermal stimuli, and naloxone reversed antinociception, indicating the involvement of the opioid system in both the peripheral and central effects of the fractions. In the writhing test, the carbohydrate fractions were the most active, inhibiting the contractions by 71 and 79 percent (B. triquetrum) and by 46 and 69 percent (B. seaforthii) at doses of 1 and 5 mg/kg, ip, respectively. Sulfated carbohydrate fractions of B. seaforthii and B. triquetrum, containing only about 5 percent protein as contaminants, are probably responsible for the antinociceptive effects of these red algae

Phaeoisaria clematidis as a cause of keratomycosis.

We report the first case of human infection by Phaeoisaria clematidis. This fungus caused a corneal ulcer in a Brazilian man who had previously suffered an eye injury. Diagnosis was established by positive direct examination and repeated cultures. The isolate was clearly resistant in vitro to the six antifungal agents tested.

[Newborn intestinal colonization by multidrug resistant enterobacteria in a neonatal unit] / Colonização intestinal de recém-natos por enterobactérias multirresistentes a antimicrobianos em unidade neonatal.

OBJECTIVE: To evaluate the occurrence of intestinal colonization in newborns by multidrug-resistant enterobacteria strains (MDRES) during hospital stay after birth. We used selective media in an attempt to determine the relationship between isolation of these strains and some of the presumed colonization risk factors. METHOD: A sequencial inclusion study of 30 newborns was carried out in the neonatal unit of the HUPE, State University Hospital, a general 600-bed tertiary care hospital. We obtained clinical and epidemiological information from medical records and collected a fecal sample from each newborn, which was plated in gentamicin (8mg/ml) medium and potassium tellurite (25mg/ml) medium. The isolated strains were biochemically identified and also submitted to tests of antimicrobial susceptibility. Nine MDRES were submitted to an assay for plasmid conjugational transfer. RESULTS: We isolated 56 distinct MDRES from 14 among 30 newborns (46.7%). Klebsiella pneumoniae was the most common bacterial species (38/56 (68%). We found statistical association between individual MDRES isolation and presence of 3 or 4 of the following colonization risk factors considered: antimicrobial use, low weight (<2.500g), more than 6 days of hospitalization and artificial milk feeding (p< 0.02). We could detect plasmid resistance transfer by bacterial conjugation for 8 among 9 MDRES. CONCLUSION: The seletive cultura media were useful to detect the high frequence of newborns colonized by MDRES in association with well established infection risk factors. We emphasize the importance of reinforcing control rules aiming at preventing intestinal colonization viewed as a risk of nosocomial infection.

Presença de pestivírus e anticorpos contra pestivírus em soros e cultivos celulares / Presence of pestiviruses and antibodies to pestiviruses in sera and cell cultures

Cultivos celulares e soro utilizados como suplemento de meios de cultivo foram examinados em busca de pestivírus ou anticorpos contra pestivírus, por técnicas de isolamento de vírus e soro-neutralizaçäo, seguidas de revelaçäo por imunoperoxidase. Sete de um total de 13 partidas de linhagens celulares, incluindo células de origem bovina (MDBK, GBK, BT), ovina (FLK) e suína (SK6, PK15), apresentavam-se contaminadas com pestivírus. Uma linhagem celular de origem de rim de coelho (RK13) näo apresentou contaminaçäo. Duas de sete partidas de cultivos primários de testículos de neonatos bovinos preparados no local também se encontraram infectados. Soros de neonatos bovinos obtidos de abatedouros continham pestivírus em oito de 80 amostras e anticorpos anti-pestivírus em 15 de 80 amostras examinadas. Soro fetal bovino adquirido comercialmente de diferentes firmas apresentou anticorpos anti-pestivírus em quatro de oito amostras examinadas

Estudo da flora fúngica da comjuntiva normal, da cana-de-açúcar e de anemófilos da regiäo canavieira de Santa Rita, Paraíba, Brasil / Study of the fungi flora in conjunctiva of sugar-cane workers and in the air and leaves of the sugar-cane plants in the Northest of Brazil

O estudo da microbiotica fúngica da conjuntiva sadia de 104 trabalhadores no corte da cana-de-açucar, no município de Santa Rita, Estado da Paraíba, apresentou como conclusöes que: 1) As culturas foram positivas em 38,5% dos indivíduos. 2) O fungo filamentoso mais comumente encontrado foi o Fusarium ssp (19,65%) sendo a espécie F. solani a mais freqüente (40,0%). Dentre as leveduras, destacou-se a Candida spp (50,0%) e a espécie C. albicans (43,58%). 3) Houve predominância dos fungos leveduriformes sobre os filamentosos. 4) A média do número de colônias por placas näo foi alta, mostrando uma microbiótica reduzida em cada trabalhador. 5) Fungos reconhecidamente patogênicos para o olho fizeram parte da microbiota fúngica. Nove gêneros diferentes foram identificados, mostrando uma microbióta variada. 6) A freqüência fúngica, quando comparada aos resultados de SCARPI, mostrou grande variaçäo. 7) A maioria dos fungos encontrados na conjuntiva foram isolados também na superfície epidérmica da folha de cana-de-açucar e colmo como também no ar, mostrando a participaçäo da contaminaçäo ambiental para conjuntiva. 8) Foi significante o aumento progressivo da freqüência fúngica em conjuntiva normal com o evoluir da idade. O estudo da microbiótica fúngica da cana-de açucar, epiderme colmo e da folha demonstrou que: 1) Os fungos isolados pertenciam a 13 gêneros. 2) O fungo mais frequentemente identificado foi o Fusarium spp., sendo o F. moniliforme na epiderme do colmo da cana, e o solani na epiderme da folha de cana os mais freqüentes. 3) Há maior variaçäo no gênereo entre os fungos presentes na folha. 4) A Candida albicans foi a levedura mais encontrada. 5) Os fungos mais freqüentemente encontrados por SCARPI, foram verificados em baixa freqüência nesse estudo e vice-versa. A análise da microbióta fúngica anemófila do canavial revelou que: 1) Identificaram-se 18 variedades de gêneros; 2) O fungo mais comumente isolado foi o Fusarium, e a espécie foi F. oxysporum; 3) A freqüência média de crescimento de fungo por placa foi alta; em 20 placas expostas ao meio ambiente, 18 mostraram crescimento fúngico; 4) Candida albicans foi a levedura identificada com maior freqüência. 5) Houve grande variedade fúngica nesse estudo, em comparaçäo com o trabalho de SCARPI

Analysis of traditional strategies to combat world hunger and their results.

The traditional strategies proposed for development of the agricultural production of the underdeveloped countries are examined in this article. It is shown that the application of these strategies in the past has proven insufficient to eliminate world hunger. The limitations of these strategies are discussed and proposals are made for achieving the agricultural development in the underdeveloped world that will be necessary to eliminate hunger in the near future.

Assessment of the world food situation-present and future.

The widespread bad harvests of 1972 in various regions of the world, the consequent reduction in grain reserves, the rapid rise in food prices almost everywhere and its impact on inflation, all have served to draw renewed attention to the problem of hunger which affects millions of human beings in the world today. During the 1974 United Nations World Food Conference many important matters relating to this problem were debated: the creation of international grain reserves; problems concerning world trade of foodstuffs; the current difficulties with certain key agricultural production factors, such as fertilizers; the necessity for organizing a worldwide information system on the situation; and prospects of various harvests and threats of famine in underdeveloped countries. It is often the case that discussion of the hunger problem does not correspond to the gravity of the crisis; true causes of the present situation are not examined, and measures are not adopted that will once and for all--for the first time--abolish hunger. In view of the prospect that the real issues are often ignored, the Transnational Institute provides this analysis in an attempt to clarify what must be done to abolish hunger in the belief that this is within the reach of humanity when and if we are determined to end the irrationalities of the present economic system and the relations of domination which some individuals and countries seek to continue.