Rumen-protected methionine supplementation improves lactation performance and alleviates inflammation during a subclinical mastitis challenge in lactating dairy cows.

This study aimed to evaluate the effects of rumen-protected Met on lactation performance, inflammation and immune response, and liver glutathione of lactating dairy cows during a subclinical mastitis challenge (SMC). Thirty-two Holstein cows (145 ± 51 DIM) were enrolled in a randomized complete block design. At -21 d relative to the SMC, cows were assigned to dietary treatments, and data were collected before and during the SMC. Cows were blocked according to parity, DIM, and milk yield and received a basal diet (17.4% CP; Lys 7.01% MP and Met 2.14% MP) plus 100 g/d of ground corn (CON; n = 16) or a basal diet plus 100 g/d of ground corn and rumen-protected Met (SM, Smartamine M at 0.09% of dietary DM; n = 16), fed as a top-dress. At 0 d, the mammary gland's rear right quarter was infused with 100,000 cfu of Streptococcus uberis (O140J). Milk yield was recorded twice daily from 0 until 3 d relative to SMC. Milk samples were collected during each milking from 0 to 3 d relative to SMC, blood samples were collected at 0, 6, 12, 24, 48, and 72 h relative to SMC. The mTOR pathway activation was assessed in immune cells in blood and milk samples by measuring quantity and phosphorylation status of mTOR-related proteins, including AKT, S6RP, and 4EBP1. For the ratio of phosphorylated to total AKT, S6RP, and 4EBP1, blood samples were collected at 0, 12, and 24 h, and milk samples at 24 h relative to SMC. Liver biopsies were performed at -10 d and 24 h relative to SMC for measurement of glutathione. Linear mixed models with repeated measures were used to analyze the results. There was a trend for greater milk yield per milking (+ 0.8 kg) and per day (+1.7 kg) after SMC in SM cows compared with CON. The DMI was not affected by dietary treatments. Reactive oxygen metabolites (ROM) were lower in SM cows than in CON. Milk somatic cell linear score was not affected by dietary treatments, and a score >4 at 24 h confirmed subclinical mastitis. The SM cows had greater milk fat percentage at 24 and 36 h post SMC, resulting in overall greater milk fat. Milk protein tended to be greater in SM cows than in CON. We observed greater liver glutathione in SM cows than in CON. Among inflammation biomarkers, ceruloplasmin was lower for SM cows compared with CON. In milk, greater pAKT:AKT and pS6RP:S6RP ratios were observed in immune cell populations from SM cows compared with CON. Blood neutrophils had a greater p4EBP1:4EBP1 ratio in SM cows compared with CON. Overall, our results show that Met supplementation during an SMC positively affected milk performance, lowered the risk of oxidative stress, and attenuated inflammation partially by increasing liver glutathione and immune cells' protein synthesis via mTOR signaling.

Fungal Planet description sheets: 1284-1382.

Novel species of fungi described in this study include those from various countries as follows: Antartica, Cladosporium austrolitorale from coastal sea sand. Australia, Austroboletus yourkae on soil, Crepidotus innuopurpureus on dead wood, Curvularia stenotaphri from roots and leaves of Stenotaphrum secundatum and Thecaphora stajsicii from capsules of Oxalis radicosa. Belgium, Paraxerochrysium coryli (incl. Paraxerochrysium gen. nov.) from Corylus avellana. Brazil, Calvatia nordestina on soil, Didymella tabebuiicola from leaf spots on Tabebuia aurea, Fusarium subflagellisporum from hypertrophied floral and vegetative branches of Mangifera indica and Microdochium maculosum from living leaves of Digitaria insularis. Canada, Cuphophyllus bondii from a grassland. Croatia, Mollisia inferiseptata from a rotten Laurus nobilis trunk. Cyprus, Amanita exilis on calcareous soil. Czech Republic, Cytospora hippophaicola from wood of symptomatic Vaccinium corymbosum. Denmark, Lasiosphaeria deviata on pieces of wood and herbaceous debris. Dominican Republic, Calocybella goethei among grass on a lawn. France (Corsica), Inocybe corsica on wet ground. France (French Guiana), Trechispora patawaensis on decayed branch of unknown angiosperm tree and Trechispora subregularis on decayed log of unknown angiosperm tree. Germany, Paramicrothecium sambuci (incl. Paramicrothecium gen. nov.) on dead stems of Sambucus nigra. India, Aureobasidium microtermitis from the gut of a Microtermes sp. termite, Laccaria diospyricola on soil and Phylloporia tamilnadensis on branches of Catunaregam spinosa. Iran, Pythium serotinoosporum from soil under Prunus dulcis. Italy, Pluteus brunneovenosus on twigs of broadleaved trees on the ground. Japan, Heterophoma rehmanniae on leaves of Rehmannia glutinosa f. hueichingensis. Kazakhstan, Murispora kazachstanica from healthy roots of Triticum aestivum. Namibia, Caespitomonium euphorbiae (incl. Caespitomonium gen. nov.) from stems of an Euphorbia sp. Netherlands, Alfaria junci, Myrmecridium junci, Myrmecridium juncicola, Myrmecridium juncigenum, Ophioceras junci, Paradinemasporium junci (incl. Paradinemasporium gen. nov.), Phialoseptomonium junci, Sporidesmiella juncicola, Xenopyricularia junci and Zaanenomyces quadripartis (incl. Zaanenomyces gen. nov.), from dead culms of Juncus effusus, Cylindromonium everniae and Rhodoveronaea everniae from Evernia prunastri, Cyphellophora sambuci and Myrmecridium sambuci from Sambucus nigra, Kiflimonium junci, Sarocladium junci, Zaanenomyces moderatricis-academiae and Zaanenomyces versatilis from dead culms of Juncus inflexus, Microcera physciae from Physcia tenella, Myrmecridium dactylidis from dead culms of Dactylis glomerata, Neochalara spiraeae and Sporidesmium spiraeae from leaves of Spiraea japonica, Neofabraea salicina from Salix sp., Paradissoconium narthecii (incl. Paradissoconium gen. nov.) from dead leaves of Narthecium ossifragum, Polyscytalum vaccinii from Vaccinium myrtillus, Pseudosoloacrosporiella cryptomeriae (incl. Pseudosoloacrosporiella gen. nov.) from leaves of Cryptomeria japonica, Ramularia pararhabdospora from Plantago lanceolata, Sporidesmiella pini from needles of Pinus sylvestris and Xenoacrodontium juglandis (incl. Xenoacrodontium gen. nov. and Xenoacrodontiaceae fam. nov.) from Juglans regia. New Zealand, Cryptometrion metrosideri from twigs of Metrosideros sp., Coccomyces pycnophyllocladi from dead leaves of Phyllocladus alpinus, Hypoderma aliforme from fallen leaves Fuscopora solandri and Hypoderma subiculatum from dead leaves Phormium tenax. Norway, Neodevriesia kalakoutskii from permafrost and Variabilispora viridis from driftwood of Picea abies. Portugal, Entomortierella hereditatis from a biofilm covering a deteriorated limestone wall. Russia, Colpoma junipericola from needles of Juniperus sabina, Entoloma cinnamomeum on soil in grasslands, Entoloma verae on soil in grasslands, Hyphodermella pallidostraminea on a dry dead branch of Actinidia sp., Lepiota sayanensis on litter in a mixed forest, Papiliotrema horticola from Malus communis, Paramacroventuria ribis (incl. Paramacroventuria gen. nov.) from leaves of Ribes aureum and Paramyrothecium lathyri from leaves of Lathyrus tuberosus. South Africa, Harzia combreti from leaf litter of Combretum collinum ssp. sulvense, Penicillium xyleborini from Xyleborinus saxesenii, Phaeoisaria dalbergiae from bark of Dalbergia armata, Protocreopsis euphorbiae from leaf litter of Euphorbia ingens and Roigiella syzygii from twigs of Syzygium chordatum. Spain, Genea zamorana on sandy soil, Gymnopus nigrescens on Scleropodium touretii, Hesperomyces parexochomi on Parexochomus quadriplagiatus, Paraphoma variabilis from dung, Phaeococcomyces kinklidomatophilus from a blackened metal railing of an industrial warehouse and Tuber suaveolens in soil under Quercus faginea. Svalbard and Jan Mayen, Inocybe nivea associated with Salix polaris. Thailand, Biscogniauxia whalleyi on corticated wood. UK, Parasitella quercicola from Quercus robur. USA, Aspergillus arizonicus from indoor air in a hospital, Caeliomyces tampanus (incl. Caeliomyces gen. nov.) from office dust, Cippumomyces mortalis (incl. Cippumomyces gen. nov.) from a tombstone, Cylindrium desperesense from air in a store, Tetracoccosporium pseudoaerium from air sample in house, Toxicocladosporium glendoranum from air in a brick room, Toxicocladosporium losalamitosense from air in a classroom, Valsonectria portsmouthensis from air in men's locker room and Varicosporellopsis americana from sludge in a water reservoir. Vietnam, Entoloma kovalenkoi on rotten wood, Fusarium chuoi inside seed of Musa itinerans, Micropsalliota albofelina on soil in tropical evergreen mixed forests and Phytophthora docyniae from soil and roots of Docynia indica. Morphological and culture characteristics are supported by DNA barcodes. Citation: Crous PW, Osieck ER, Jurjevic Z, et al. 2021. Fungal Planet description sheets: 1284-1382. Persoonia 47: 178-374. https://doi.org/10.3767/persoonia.2021.47.06.

Fungal Planet description sheets: 1284-1382.

Novel species of fungi described in this study include those from various countries as follows: Antartica, Cladosporium austrolitorale from coastal sea sand. Australia, Austroboletus yourkae on soil, Crepidotus innuopurpureus on dead wood, Curvularia stenotaphri from roots and leaves of Stenotaphrum secundatum and Thecaphora stajsicii from capsules of Oxalis radicosa. Belgium, Paraxerochrysium coryli (incl. Paraxerochrysium gen. nov.) from Corylus avellana. Brazil, Calvatia nordestina on soil, Didymella tabebuiicola from leaf spots on Tabebuia aurea, Fusarium subflagellisporum from hypertrophied floral and vegetative branches of Mangifera indica and Microdochium maculosum from living leaves of Digitaria insularis. Canada, Cuphophyllus bondii from a grassland. Croatia, Mollisia inferiseptata from a rotten Laurus nobilis trunk. Cyprus, Amanita exilis on calcareous soil. Czech Republic, Cytospora hippophaicola from wood of symptomatic Vaccinium corymbosum. Denmark, Lasiosphaeria deviata on pieces of wood and herbaceous debris. Dominican Republic, Calocybella goethei among grass on a lawn. France (Corsica), Inocybe corsica on wet ground. France (French Guiana), Trechispora patawaensis on decayed branch of unknown angiosperm tree and Trechispora subregularis on decayed log of unknown angiosperm tree. Germany, Paramicrothecium sambuci (incl. Paramicrothecium gen. nov.) on dead stems of Sambucus nigra. India, Aureobasidium microtermitis from the gut of a Microtermes sp. termite, Laccaria diospyricola on soil and Phylloporia tamilnadensis on branches of Catunaregam spinosa. Iran, Pythium serotinoosporum from soil under Prunus dulcis. Italy, Pluteus brunneovenosus on twigs of broadleaved trees on the ground. Japan, Heterophoma rehmanniae on leaves of Rehmannia glutinosa f. hueichingensis. Kazakhstan, Murispora kazachstanica from healthy roots of Triticum aestivum. Namibia, Caespitomonium euphorbiae (incl. Caespitomonium gen. nov.) from stems of an Euphorbia sp. Netherlands, Alfaria junci, Myrmecridium junci, Myrmecridium juncicola, Myrmecridium juncigenum, Ophioceras junci, Paradinemasporium junci (incl. Paradinemasporium gen. nov.), Phialoseptomonium junci, Sporidesmiella juncicola, Xenopyricularia junci and Zaanenomyces quadripartis (incl. Zaanenomyces gen. nov.), from dead culms of Juncus effusus, Cylindromonium everniae and Rhodoveronaea everniae from Evernia prunastri, Cyphellophora sambuci and Myrmecridium sambuci from Sambucus nigra, Kiflimonium junci, Sarocladium junci, Zaanenomyces moderatricis-academiae and Zaanenomyces versatilis from dead culms of Juncus inflexus, Microcera physciae from Physcia tenella, Myrmecridium dactylidis from dead culms of Dactylis glomerata, Neochalara spiraeae and Sporidesmium spiraeae from leaves of Spiraea japonica, Neofabraea salicina from Salix sp., Paradissoconium narthecii (incl. Paradissoconium gen. nov.) from dead leaves of Narthecium ossifragum, Polyscytalum vaccinii from Vaccinium myrtillus, Pseudosoloacrosporiella cryptomeriae (incl. Pseudosoloacrosporiella gen. nov.) from leaves of Cryptomeria japonica, Ramularia pararhabdospora from Plantago lanceolata, Sporidesmiella pini from needles of Pinus sylvestris and Xenoacrodontium juglandis (incl. Xenoacrodontium gen. nov. and Xenoacrodontiaceae fam. nov.) from Juglans regia. New Zealand, Cryptometrion metrosideri from twigs of Metrosideros sp., Coccomyces pycnophyllocladi from dead leaves of Phyllocladus alpinus, Hypoderma aliforme from fallen leaves Fuscopora solandri and Hypoderma subiculatum from dead leaves Phormium tenax. Norway, Neodevriesia kalakoutskii from permafrost and Variabilispora viridis from driftwood of Picea abies. Portugal, Entomortierella hereditatis from a biofilm covering a deteriorated limestone wall. Russia, Colpoma junipericola from needles of Juniperus sabina, Entoloma cinnamomeum on soil in grasslands, Entoloma verae on soil in grasslands, Hyphodermella pallidostraminea on a dry dead branch of Actinidia sp., Lepiota sayanensis on litter in a mixed forest, Papiliotrema horticola from Malus communis, Paramacroventuria ribis (incl. Paramacroventuria gen. nov.) from leaves of Ribes aureum and Paramyrothecium lathyri from leaves of Lathyrus tuberosus. South Africa, Harzia combreti from leaf litter of Combretum collinum ssp. sulvense, Penicillium xyleborini from Xyleborinus saxesenii, Phaeoisaria dalbergiae from bark of Dalbergia armata, Protocreopsis euphorbiae from leaf litter of Euphorbia ingens and Roigiella syzygii from twigs of Syzygium chordatum. Spain, Genea zamorana on sandy soil, Gymnopus nigrescens on Scleropodium touretii, Hesperomyces parexochomi on Parexochomus quadriplagiatus, Paraphoma variabilis from dung, Phaeococcomyces kinklidomatophilus from a blackened metal railing of an industrial warehouse and Tuber suaveolens in soil under Quercus faginea. Svalbard and Jan Mayen, Inocybe nivea associated with Salix polaris. Thailand, Biscogniauxia whalleyi on corticated wood. UK, Parasitella quercicola from Quercus robur. USA, Aspergillus arizonicus from indoor air in a hospital, Caeliomyces tampanus (incl. Caeliomyces gen. nov.) from office dust, Cippumomyces mortalis (incl. Cippumomyces gen. nov.) from a tombstone, Cylindrium desperesense from air in a store, Tetracoccosporium pseudoaerium from air sample in house, Toxicocladosporium glendoranum from air in a brick room, Toxicocladosporium losalamitosense from air in a classroom, Valsonectria portsmouthensis from air in men's locker room and Varicosporellopsis americana from sludge in a water reservoir. Vietnam, Entoloma kovalenkoi on rotten wood, Fusarium chuoi inside seed of Musa itinerans, Micropsalliota albofelina on soil in tropical evergreen mixed forests and Phytophthora docyniae from soil and roots of Docynia indica. Morphological and culture characteristics are supported by DNA barcodes. Citation: Crous PW, Osieck ER, Jurjevic Z, et al. 2021. Fungal Planet description sheets: 1284-1382. Persoonia 47: 178-374. https://doi.org/10.3767/persoonia.2021.47.06.

Structural, bonding, and electronic properties of the hexagonal ferroelectric and paraelectric phases of LuMnO3 compound: a density functional theory study.

We have investigated the structural, bonding, and electronic properties of both ferroelectric (FE) and paraelectric (PE) phases of the hexagonal LuMnO3 compound using calculations based on density functional theory. The structural properties have been determined by employing the generalized gradient approximation with Perdew-Burke-Ernzerhof and Wu-Cohen parameterization. The bonding and electronic properties have been treated by recently developed modified Becke-Johnson exchange potential, which succeeded to open a band gap for both PE and FE phases, in agreement with experimental predictions. The Bader's topological analysis of electronic density showed that the character of the Lu-O axial bonds changes when the crystal exhibits the PE → FE structural transition. This fact is in agreement with experimental findings. The covalent character of the Lu-O bond significantly increases due to orbital hybridization between the Lu 5dz(2) and O 2pz-states. This bonding mechanism causes the ferroelectricity in the hexagonal LuMnO3 compound.

Accumulation and decomposition of cultural residues of Theobroma grandiflorum, Paullinia cupana, Bixa orellana and forest in the southern region of Amazonas.

The litter deposited on the soil surface at various stages of decomposition is important for primary productivity that impacts the microbial communities and soil carbon storage. The objective of this study was to evaluate the accumulation and decomposition of cultural residues of Theobroma grandiflorum (Willd. ex. Spreng) Schum, Paullinia cupana (Mart.) Ducke, Bixa orellana L., and forest in the Amazon region. The study was carried out in the São Francisco settlement, Canutama in the south of Amazonas, in a randomized block experimental design, and the treatments consisted of four areas with different crops: 1 - P. cupana; 2 - T. grandiflorum; 3 - B. orellana; 4 - Native woodland area (forest), in time subdivided plots: 7, 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, and 330 days after the distribution of the bags in the field, all with four repetitions. To evaluate the contribution and fractions of litter, conical collectors were used in each area, and collections were performed monthly in the period from March 2020 to February 2021. The estimate of the decomposition rate of the litter was done by quantifying the loss of mass, using litter bags, which allow for a direct analysis of the rate of decay over time. The forest and P. cupana environments presented the highest litter production, and greater deposition when compared to environments cultivated with T. grandiflorum and B. orellana. The forest and B. orellana areas showed the highest speed of decomposition, while the opposite situation occurred under T. grandiflorum and P. cupana cultivation.

The nematode community in the Atlantic rainforest lizard Enyalius perditus Jackson, from south-eastern Brazil.

Studies focusing on communities of helminths from Brazilian lizards are increasing, but there are many blanks in the knowledge of parasitic fauna of wild fauna. This lack of knowledge hampers understanding of ecological and parasitological aspects of involved species. Moreover, the majority of research has focused on parasitic fauna of lizards from families Tropiduridae and Scincidae. Only a few studies have looked at lizards from the family Leiosauridae, including some species of Enyalius. This study presents data on the gastrointestinal parasite fauna of Enyalius perditus and their relationships with ecological aspects of hosts in a disturbed Atlantic rainforest area in the state of Minas Gerais, south-eastern Brazil. Two nematode species, Oswaldocruzia burseyi [(Molineidae) and Strongyluris oscari (Heterakidae) were found. Nematode species showed an aggregated distribution in this host population, with O. burseyi being more aggregated than S. oscari. The present study extends the range of occurrence of O. burseyi to the Brazilian continental area.

Effect of tooth bleaching on bond strength of enamel-dentin cavities restored with silorane- and dimethacrylate-based materials.

The aim of this study was to evaluate the influence of tooth bleaching on the push-out bond strength of a composite resin based on dimethacrylates and silorane to cavities that involve both enamel and dentin. A total of 80 bovine incisors were sectioned on the buccal surface to obtain specimens (10 × 10 mm) presenting enamel and dentin (1-mm thick each substrate). The specimens were randomly distributed into eight groups (n=10), according to the bleaching protocol (1--none; 2--10% carbamide peroxide [CP] for 21 days, six hours each day; 3--three applications of 35% hydrogen peroxide [HP] in 15-minute sessions, one session every seven days for three weeks; 4--10% CP for 18 days, six hours each day + three applications of 35% HP in 15-minute sessions, one session every seven days for three weeks) and the restorative system applied (Adper Single Bond 2 + Filtek Supreme; Filtek Silorane adhesive and composite resin). After treatment, cavities were made (1.2-mm diameter on dentin; 1.5-mm diameter on enamel) with a diamond bur. At 24 hours after restoration, a push-out bond strength test was performed at a crosshead speed of 0.5 mm/min. The bleaching treatments did not significantly affect the bond strengths of either restorative system to enamel-dentin. Regardless of the bleaching treatment, the dimethacrylate-based resin system exhibited significantly higher bond strengths to enamel-dentin than did the silorane-based system.

Effect of preheating resin composite and light-curing units on the microleakage of Class II restorations submitted to thermocycling.

This in vitro study evaluated microleakage in Class II cavities restored with dental composite and varying light-curing units and the temperature of the composite when subjected to a thermocycling test. Ninety cavities were prepared on the proximal surfaces of bovine teeth and randomly divided according to the light-curing mode (QTH-420 mW/cm2, LED 2nd generation-1100 mW/cm2, or LED 3rd generation-700 mW/cm2) and temperature of the resin composite (23°C, 54°C and 60°C). Following the restorative procedures and thermocycling, the samples were immersed in methylene blue for 12 hours. The samples were ground and the powder prepared for analysis in an absorbance spectrophotometer. All the results were statistically analyzed using the nonparametric tests of Kruskal-Wallis and Dunn (p ≤ 0.05). The results showed that there was no statistical difference between the light-curing modes at a temperature of 23°C. For 54°C, QTH showed a microleakage mean that was significantly lower than those of the LED groups, and for 60°C, QTH had a microleakage mean significantly lower than that of the LED 2nd generation group. There was no statistical difference between the temperatures of the resin composite when LEDs were used. For QTH, 54°C showed statistically lower microleakage than 23°C. The group preheated to 60°C showed no difference when compared to the group heated to 23°C. Preheating the resin composite (54°C and 60°C) did not improve the microleakage means when high-irradiance LED was used; however, it decreased the microleakage means when a QTH with low irradiance was used.

Effect of eugenol exposure time and post-removal delay on the bond strength of a self-etching adhesive to dentin.

OBJECTIVES: This study evaluated the effects of the exposure time of eugenol-based provisional restorative material and the time elapsed between the provisional material removal and the adhesive procedure on the bond strength of the composite to dentin. MATERIALS AND METHODS: Human third molars were sectioned into two halves that were enclosed in resin cylinders. The cavities were prepared over the buccal/lingual faces with diamond burs. Zinc oxide and eugenol (ZOE) provisional material was inserted into cavities and left for 24 hours, 7 days or 14 days. The cavities not restored with ZOE were used as controls. After ZOE removal or over fresh dentin (control), self-etching Adper SE Plus was applied immediately, after a 7- or 14-day delay. The cavity was restored with non-eugenol provisional material during this delay period. Cylinders of resin cement were built-up over the hybridized dentin. A shear load was applied to the cylinders at a crosshead speed of 0.5 mm/minute until failure. The data were statistically analyzed using two-way ANOVA and Tukey's tests (α=0.05). RESULTS: Using IRM as a provisional restoration for 24 hours followed by its removal and immediate adhesive application resulted in the lowest values of shear bond strength. There was no significant difference between the other experimental conditions. CONCLUSIONS: The use of IRM for 24 hours adversely affected the shear bond strength of a self-etching adhesive to dentin. The recovery of the proper bond strength occurred one week after IRM removal.

Teschovirus and other swine and human enteric viruses in Brazilian watersheds impacted by swine husbandry.

Several emerging viral agents related to gastroenteritis are distributed in human and animal populations and may contaminate the environment due to anthropic activities. The objective of this study was to analyze the seasonal contamination by enteric virus and coliforms in water from streams in the Vale do Taquari, draining a large number of pig farms. Microbiological contamination was evidenced by the detection of total and thermotolerant coliforms, reaching their peak in December. Hepatitis E virus (HEV), Enterovirus-G (EV-G) genome, and Sapelovirus-A (SV-A) genome were not detected. On the other hand, Rotavirus (RV) was detected in 3% (1/32) of the samples, whereas Teschovirus-A (PTV) was detected in 6% (2/32). This is the first detection of PTV in environmental samples in Brazil, pointing that the virus is being shedded from swine herds to watersheds. Human mastadenovirus (HAdV) was the most frequent detected viral agent in 9.3% (3/32) with values of 2.54 × 105, 7.13 × 104, and 3.09 × 105 genome copies/liter (gc/L). The circulation of coliforms and viral pathogens is noticeable due to anthropic activities and to the management of animal waste from the pig farming. In this way, enteric viruses can assist in monitoring the quality of watersheds and in tracking sources of contamination.

Impact of 24-h cooling prior to freezing on the survival of domestic cat (Felis catus) epididymal sperm.

The aim of this study was to investigate the impact of a 24-h cooling period prior to freezing on domestic cat epididymal sperm viability. Fifteen tomcats were submitted to routine orchiectomy and sperm samples were retrieved from both epididymides in a Tris-glucose-20% egg yolk extender. For each tomcat, the diluted sperm was split into two equal volumes and cooled to 5 degrees C at a rate of 0.5 degrees C/min; one sample for 60 min (control) and the other for 24 h (cooled). After the cooling period, samples from both groups were frozen using an identical freezing protocol. Sperm samples were evaluated in three different periods: immediately after harvesting, after cooling at 5 degrees C for 24 h (cooled group) and after freezing-thawing of control and cooled groups. Evaluations consisted of sperm motility and progressive status, sperm morphology and plasma membrane integrity (PMI) using two fluorescent probes. After cooling for 24 h, a decrease (p < 0.05) in sperm motility, progressive status and PMI was observed when compared to sperm samples immediately after collection. Comparing the results obtained after thawing, no difference (p < 0.05) was found regarding sperm motility, progressive status, PMI and sperm morphology between control and cooled groups. The results from the present study show that cooling cat epididymal spermatozoa at 5 degrees C for 24 h prior to freezing does not lead to major damage of spermatozoa impairing the freeze-thaw process.

Osmotic modulation of chromatin impacts on efficiency and kinetics of cell fate modulation.

Chromatin structure is a major regulator of transcription and gene expression. Herein we explore the use of osmotic modulation to modify the chromatin structure and reprogram gene expression. In this study we use the extracellular osmotic pressure as a chromatin structure and transcriptional modulator. Hyposmotic modulation promotes chromatin loosening and induces changes in RNA polymerase II (Pol II) activity. The chromatin decondensation opens space for higher amounts of DNA engaged RNA Pol II. Hyposmotic modulation constitutes an alternative route to manipulate cell fate decisions. This technology was tested in model protocols of induced pluripotency and transdifferentiation in cells growing in suspension and adherent to substrates, CD34+ umbilical-cord-blood (UCB), fibroblasts and B-cells. The efficiency and kinetics of these cell fate modulation processes were improved by transient hyposmotic modulation of the cell environment.

Author Correction: Osmotic modulation of chromatin impacts on efficiency and kinetics of cell fate modulation.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Vitamin K in colostrum and mature human milk over the lactation period--a cross-sectional study.

Vitamin K was quantitated in the milk of four groups of 15 mothers from 1 d to 6 mo postpartum in a cross-sectional study. Concentrations were 7.52 +/- 5.90 and 6.36 +/- 5.32 nmol/L (3.39 +/- 2.66 and 2.87 +/- 2.40 micrograms/L) in colostrum and mature milk, respectively. Differences between colostrum and mature milk or among samples of mature milk collected at 1, 3, and 6 mo were not statistically significant. Because of significantly increased volumes of milk over the lactation period, approximately twice as much vitamin K was delivered in mature milk as in colostrum. Within normal ranges, concentrations of vitamin K in milk were not predicted by dietary intake of vegetables or fat. Vitamin K was correlated with fat in colostrum and was localized in the lipid core of the milk fat globule but was not associated with membranes. Vitamin K in human milk is insufficient to meet recommended intakes for infants aged less than 6 mo. Population and clinical studies are needed to assess the vitamin K status of exclusively breast-fed infants and to evaluate current recommendations.

Plasma response to oral beta-carotene in Guatemalan schoolchildren.

The response to oral doses of beta-carotene (0 mg, n = 10; 15 mg, n = 20; and 30 mg, n = 21) was studied in 51 Guatemalan children aged 8-15 y, with mean fasting plasma retinol concentrations of 1.72 +/- 0.38 mumol/L. Beta-carotene was delivered with a chocolate drink containing 8.4 g fat. Serial blood sampling was performed at intervals up to 48 h. Circulating retinol concentrations remained relatively constant. The maximum increases in plasma beta-carotene after the 30- and 15-mg doses for all subjects occurred at 24 h and were 0.29 and 0.23 mumol/L, respectively. Time of maximum increase for individuals varied and average maxima over the 48-h period for individuals were 0.13 and 0.26 mumol/L for the 15- and 30-mg-treatment groups, respectively. Increased plasma beta-carotene concentrations were not predicted by recent intake of dietary vitamin A, fasting plasma concentrations, or anthropometric measurements.

Quantitation of vitamin K in human milk.

A quantitative method was developed for the assay of vitamin K in human colostrum and milk. The procedure combines preparative and analytical chromatography on silica gel in a nitrogen atmosphere followed by reversed phase high performance liquid chromatography (HPLC). Two HPLC steps were used: gradient separation with ultraviolet (UV) detection followed by isocratic separation detected electrochemically. Due to co-migrating impurities, UV detection alone is insufficient for identification of vitamin K. Exogenous vitamin K was shown to equilibrate with endogenous vitamin K in the samples. A statistical method was incorporated to control for experimental variability. Vitamin K1 was analyzed in 16 pooled milk samples from 7 donors and in individual samples from 15 donors at 1 month post-partum. Vitamin K1 was present at 2.94 +/- 1.94 and 3.15 +/- 2.87 ng/mL in pools and in individuals, respectively. Menaquinones, the bacterial form of the vitamin, were not detected. The significance of experimental variation to studies of vitamin K in individuals is discussed.

Increased creatine kinase and spontaneous activity on electromyography, in amyotrophic lateral sclerosis.

INTRODUCTION: Mild to moderate elevation of muscle creatine kinase (CK) is commonly observed in amyotrophic lateral sclerosis (ALS). Although the determinants of increased the CK in ALS remain uncertain, we hypothesize that fasciculations and muscle denervation can be involved by damaging the muscle fibre. PATIENTS AND METHODS: We studied 87 ALS patients in whom CK determination was performed. In 47, a standardized EMG investigation was performed. In 22 patients a second CK determination was performed a mean of 5 months later. CK values were compared between different patients arranged in groups as determined by the number of regions with fasciculation as detected on the clinical examination, and the number of muscles with fasciculation or with fibrillation potentials as observed on EMG. RESULTS: 43% of our population had an increased CK value. Four out of 5 patients with suspected ALS had an increased CK value. The number of patients with increased CK value was not different between sexes, or between bulbar and spinal-onset patients. CK value was not related with disease duration, and did not change at the second measurement. CK value was not different between the groups studied. CONCLUSION: The fasciculations,and the signs of denervation on EMG, are not determinants for high CK values in ALS patients, which are still unknown. Increased CK can be useful in the differential diagnosis of patients with lower motor neuron disorders.

[Limitations and alternatives to the implementation of a program for drug users at risk of HIV infection using the Social Network Approach]. / Limites e alternativas para a implementação de um programa para dependentes químicos em risco para infecção pelo HIV utilizando o conceito de Rede Social.

The authors describe the development of a preventive program focused on intravenous drug users at risk of HIV infection, using the Social Network Approach as the intervention model. The authors describe the project's steps in a large university hospital in southern Brazil, emphasizing the unique methods and techniques developed by the treatment staff. Problems encountered during the project development are discussed, aimed at identifying the reasons why the program only achieved partial success. The authors identify critical issues, such as the use of a new technique not previously tried in Brazil, difficulties in maintaining IV drug users in treatment, lack of infrastructure for walk-in treatment, and the challenge of motivating staff and patients to continue treatment. The authors conclude by listing suggestions aimed at facilitating the development of new projects based on the same conceptual model.

Cervical enamel projection related to furcation involvement.

Reviewing the literature the authors show the importance of cervical enamel projections (CEP) in the involvement of molar furcation. The dento-gingival relationship of CEP structure is peculiar for not having connective attachment but only long junctional epithelium. In numerous studies, most mandibular and maxillary molars presented CEP, mainly of little extension, with an incidence ranging from 8.8 to 87.4%. The studies showed a relationship between cervical enamel projection and the presence of inflammatory periodontal disease, and furcation involvement of molars. These data suggest that a detailed clinical trial must be carried out as well as early diagnosis of periodontal disease at the region of furcation.

Scanning electron microscopy study of the effect of tetracycline HCl on smear layer removal and fibrin network formation.

Scanning electron microscopy (SEM) was used to evaluate root surface characteristics of human teeth affected with periodontitis following periodontal instrumentation and topical application of tetracycline HCl (TTC-HCl; pH 1.6; 4 min). Specimens were randomly assigned to periodontal instrumentation alone (control 1); periodontal instrumentation plus TTC-HCl (test 1); periodontal instrumentation plus trypsin solution after extraction (control 2); and periodontal instrumentation plus TTC-HCl plus trypsin solution after extraction (test 2). Tetracycline solution was applied with a cotton pellet. Twenty-two single root periodontitis affected human teeth scheduled for extraction were selected. Mucoperiosteal flaps were raised, root surfaces were mechanically and chemically treated, flaps were repositioned and maintained in place for 20 min. Teeth were extracted, rinsed and placed in cold phosphate buffer solution (PBS) and control 2 and test 2 groups were treated with trypsin solution. Specimens were examined using SEM. Smear layer was successfully removed, exposing dentinal tubules; however, fibrin network formation in situ was not improved by application of TTC-HCl.