Production of mycelial biomass, proteases and protease inhibitors by Ganoderma lucidum under different submerged fermentation conditions.

Ganoderma lucidum is a medicinal mushroom widely recognized as a source of biomolecules with pharmacological properties, however, little is known about the factors that influence the synthesis of bioactive proteins by this fungus when cultivated under submerged fermentation. The objective of this work was to evaluate the production of mycelial biomass and intracellular proteases and protease inhibitors by G. lucidum cultivated under different submerged fermentation conditions. The cultivation was carried out in a medium composed of glucose (10 or 20 g.L-1), soy peptone (2.5 or 5 g.L-1) and yeast extract (5 g.L-1), with incubation under agitation (120 rpm) and non-agitation, totaling 8 experimental conditions. Biomass production was determined from the dry weight, while glucose consumption was estimated by quantification of reducing sugars. The proteins were extracted in NaCl (0.15 M), and the protein extracts were submitted to protein quantification by the Bradford method, total proteolytic activity using azocasein, caseinolytic and fibrinolytic activity in Petri dishes, activity of serine (trypsin and chymotrypsin) and cysteine (papain) protease inhibitors. Cultivation in agitated condition showed higher biomass production with a maximum value of 7 g.L-1, in addition to higher activities of trypsin, chymotrypsin and papain inhibitors, with 154 IU.mg-1, 153 IU.mg-1 e 343 IU.mg-1 of protein, respectively. The non-agitated condition showed a greater potential for obtaining proteins, total proteases, caseinolytic and fibrinolytic enzymes, with maximum values of 433 mg.g-1 of extract, 71 U.mL-1 of extract, 63.62 mm2 and 50.27 mm2, respectively. Thus, a medium composed of soy peptone, yest extract and glucose in a 1:2:4 proportion is recommended, under agitation to produce protease inhibitors, and the non-agitated condition when the target is, mainly caseinolytic and fibrinolytic enzymes.

Culicoides spp. found near Lusitano stud farms in mainland Portugal which may contribute for IBH studies.

Insect Bite Hypersensitivity (IBH) is a common cutaneous disease, affecting a large number of horses worldwide. Several studies have identified Culicoides spp. saliva as a clinically relevant allergen source. The prevalence of IBH in Portugal, particularly in Lusitano horses, is still not known. However, the environmental characteristics of the national territory are favorable to the activity of Culicoides, and several species of this genus can be found, namely C. imicola and C. obsoletus/C. scoticus. In this study we characterized the Culicoides population present in Lusitano stud farms with a history of IBH. Thirteen stud farms with Lusitano horses were selected in several regions of mainland Portugal for having a previous history of IBH-affected horses, with a minimum of 5 affected horses. Culicoides were collected in May and June 2016 using OVI traps, placed in these stud farms, and we were able to identify several Culicoides species. We could also verify that C. obsoletus/C. scoticus, and C. imicola were the ones most frequently found, but other species like C. pulicaris were also found.

Distinct chromatin structures at the monoamine oxidase-A promoter correlate with allele-specific expression in SH-SY5Y cells.

Monoamine oxidase-A (MAOA) metabolises monoamines and is implicated in the pathophysiology of psychiatric disorders. A polymorphic repetitive DNA domain, termed the uVNTR (upstream variable number tandem repeat), located at the promoter of the MAOA gene is a risk factor for many of these disorders. MAOA is on the X chromosome suggesting gender could play a role in regulation. We analysed MAOA regulation in the human female cell line, SH-SY5Y, which is polymorphic for the uVNTR. This heterozygosity allowed us to correlate allele-specific gene expression with allele-specific transcription factor binding and epigenetic marks for MAOA. Gene regulation was analysed under basal conditions and in response to the mood stabiliser sodium valproate. Both alleles were transcriptionally active under basal growth conditions; however, the alleles showed distinct transcription factor binding and epigenetic marks at their respective promoters. Exposure of the cells to sodium valproate resulted in differential allelic expression which correlated with allele-specific changes in distinct transcription factor binding and epigenetic marks at the region encompassing the uVNTR. Biochemically our model for MAOA promoter function has implications for gender differences in gene × environment responses in which the uVNTR has been implicated as a genetic risk.

Clinical Evolution of Patients Using Ventricular Assist Devices as a Bridge for Transplantation.

OBJECTIVE: To evaluate the clinical evolution of patients with implantation of ventricular assist device (VAD) and identify the intervening factors for death. METHODS: This analytical, retrospective study was carried out in a public reference hospital in cardiopulmonary diseases located in northeastern Brazil. The study population encompassed the medical records of 16 patients who underwent VAD implantation. Data collection took place from January to August 2016, through the consultation of medical records. Descriptive analysis, odds ratio, and the Fisher's Exact, Wilcoxon, Friedman and t-tests were used to analyze the data. RESULTS: All patients experienced complications during the use of the device, with bleeding being the main cause (11 [68.8%]). There was a significant decrease in noradrenaline (P = .025), milrinone (Primacor; P = .007), and dobutamine (P = .046) flow rates with the clinical evolution of patients. Regarding hematologic parameters, the use of VAD promoted a significant improvement in hemoglobin (P < .001), hematocrit (P = .003), activated partial thromboplastin time (P = .013), and fibrinogen (P = .049) values at the 3 time points analyzed. Regarding the clinical outcome of the patients, the majority (10 [62.5%]) underwent cardiac transplantation. CONCLUSIONS: This study allowed for better knowledge of the clinical evolution of patients with VAD implantation, highlighting the benefits of this type of device as a bridge for heart transplantation.

A TOMM40 poly-T variant modulates gene expression and is associated with vocabulary ability and decline in nonpathologic aging.

The Translocase of Outer Mitochondrial Membrane 40 Homolog and Apolipoprotein E (TOMM40-APOE) locus has been associated with a number of age-related phenotypes in humans including nonpathologic cognitive aging, late-onset Alzheimer's disease, and longevity. Here, we investigate the influence of the TOMM40 intron 6 poly-T variant (rs10524523) on TOMM40 gene expression and cognitive abilities and decline in a cohort of 1613 community-dwelling elderly volunteers who had been followed for changes in cognitive functioning over a period of 14 years (range = 12-18 years). We showed that the shorter length poly-T variants were found to act as a repressor of luciferase gene expression in reporter gene constructs. Expression was reduced to approximately half of that observed for the very long variant. We further observed that the shorter poly-T variant was significantly associated with reduced vocabulary ability and a slower rate of vocabulary decline with age compared to the very long poly-T variants. No significant associations were observed for memory, fluid intelligence or processing speed, although the direction of effect, where the short variant was correlated with reduced ability and slower rate of decline was observed for all tests. Our results indicate that the poly-T variant has the ability to interact with transcription machinery and differentially modulate reporter gene expression and influence vocabulary ability and decline with age.

Candida colonisation as a source for candidaemia.

Candida spp. are important healthcare-associated pathogens. Identifying the source of infection is important for prevention and control strategies. The objective of this study was to evaluate candida colonisation sites as potential sources for candidaemia. Sixty-three consecutive patients with a positive blood culture for candida were included. Surveillance cultures were collected from urine, rectum, oropharynx, skin, intravascular catheter tip and skin around catheter. Molecular typing was performed when the same species of candida was isolated from blood and surveillance sites of a patient. C. albicans was associated with 42% of candidaemias, C. parapsilosis 33%, C. tropicalis 16% and C. guilliermondii, C. krusei, C. glabrata, C. holmii and C. metapsilosis were all 2% each. Six of 10 C. parapsilosis catheter tip isolates were indistinguishable from corresponding blood isolates (all in neonates). C. albicans isolates from blood were indistinguishable from corresponding gastrointestinal tract isolates in 13 of 26 patients and from catheter tip isolates in two patients. In conclusion, the results suggest that gastrointestinal colonisation is the probable source of C. albicans candidaemia and C. parapsilosis is exogenous.

Filling in at partially deafferented visual cortex.

METHODS: Perceptual visual filling in at the corresponding area of toxoplasmic retinochoroiditis scotomas in the fellow eye was studied in 14 patients. Targets were programmed to appear inside the scotoma corresponding area in the fellow eye, and at two control points. RESULTS: The results showed faster perceptual filling-in latencies (p<0.0001) in the scotoma corresponding area than at the control points. CONCLUSIONS: Perceptual filling in is facilitated at the corresponding area of retinal scotomas. The neural mechanisms of this visual facilitation might be similar to those found following somatosensory cortex partial deafferentation.

Color vision discrimination in the capuchin monkey Cebus apella: evidence for trichromaticity.

Primates display significant differences in color vision. The purpose of this study was to assess the ability of capuchin monkeys in discriminating chromatic and achromatic Munsell color chips through behavioral tests. The subjects were trained in a simple and reverse discrimination learning procedure. All subjects were capable of discriminations along five color categories investigated. The results are discussed in terms of the hypothesis of male dichromatism in New World monkeys, the role of color vision in adaptation to feeding ecology, as well as to aspects regarding primate evolution.

Seroprevalência da infecçäo pelo HIV-1 em gestantes atendidas no IMIP, Recife - Brasil / Seroprevalence of HIV infection among pregnant women attending in the IMIP, Recife-Brasil

The number of HIV infected heterosexual has been steadily increasing in Brazil. This fact was followed by a decline on the ratio of male/female HIV infection every year. We have examined the seroprevalence of HIV infection among pregnant women attending an antenatal clinic in the Instituto Materno Infantil de Pernambuco (IMIP) at Recife, North-eastern Brazil. This study was performae as an anonymous sentinel surveillance. The collected blood samples were tested for HIV infection by two different types of ELISA, and if positive, further analysis by IIA was submitted. The subjects were arranged in four age groups with the respective proportion: group l (<15 years)-0.9 per cent, group II (15 to 20 years)-31.6 per cent, group III (21 to 34 years)-61.7 per cent and group IV (>35 years)- 5,8 per cent. It was found an HIV-1 seroprevalence of 0.1 per cent (95 per cent confidence interval, -0.l to + 0,3) This percentage represents one single HIV positive woman included in age group III. his low HIV infection seroprevalence is suitable with a low risk population (pregnant women) in a low prevalence area. However, we became concerned with the great propottion of pregnant woman under 21 years (32,5 per cent) and with the implications of how to reinforce effectively HIV preventions of how to reinforce effectively HIV prevention strategies to this population

Relative sizes of cortical visual areas in marmosets: functional and phylogenetic implications.

Visual areas V1, V2 and MT (V5) were identified through myeloarchitectonic criteria and their sizes estimated in a flattened map of caudal cerebral cortex in the marmoset Callithrix penicillata. The ratio MT/V1 in this species is similar to values reported for other species of primates, but the ratio V2/V1 in Callithrix penicillata is smaller than that in macaques and larger than that in Aotus. The possible implications of these results are discussed.

Ganglion cell size and distribution in the retina of the two-toed sloth (Choloepus didactylus L.).

The distribution of ganglion cell densities and sizes was studied in Nissl-stained flat-mount retinae of the two-toed sloth. The area centralis, a weak specialization with low ganglion cell density, is located in the temporal retina close to the center of the eye. The presence of a visual streak was noted. The distribution of different ganglion cell sizes was approximately equal throughout the retina. Although the retinal organization differs from that of the closely related three-toed sloth, the presumed function of retinal specializations in both species is to guide limb movements by permitting visualization of the branch along which the animal is climbing.

Ganglion cell size and distribution in the retina of the two-toed sloth (Choloepus didactylus L.)

The distribution of ganglion cell densities and sizes was studied in Nissl-stained flat-mount retinae fo the two-toed sloth. The area centrailis, a weak specializtion with low ganglion cell density, is located in the temporal retina close to the center of the eye. The presence of a visual treak was nsoted. The distribution of different ganglion cell sizes was approximately equal throughout the retina. Although the retinal organization differs from that of the closely related three-toed sloth, presumed function of retinl specializations in both species is to guide limb movements by permiting visualization of the branch along which the animal is climbing

Distribution of motoneurones innervating extraocular muscles in the brain of the marmoset (Callithrix jacchus).

Extraocular muscle motoneurones were localised in the oculomotor nucleus (ON), trochlear nucleus (TN) and abducens nucleus (AN) in the marmoset brain using the horseradish peroxidase (HRP) retrograde labelling technique. HRP pellets injected into individual extraocular muscles revealed one or more groups of labelled neurones occupying discrete loci within these nuclei. Relatively little overlap of motoneurone pools was observed, except in the case of the inferior oblique and superior rectus muscles. Injections of HRP into the medial rectus muscle revealed three separate populations of labelled cells in the ipsilateral ON. Motoneurones innervating the inferior rectus muscle were mainly localised in the lateral somatic cell column of the ipsilateral ON. A second smaller grouping was observed in the medial longitudinal fasciculus. The inferior oblique muscle motoneurones were localised in the ipsilateral medial somatic cell column intermingled with motoneurones supplying the superior rectus muscle of the opposite eye. The superior oblique muscle motoneurones occupied the entire TN and the lateral rectus muscle motoneurones the AN. It was concluded that the organisation of nuclei and subnuclei responsible for controlling the extraocular muscles in the marmoset is broadly similar to that of other primates.

Unusual distribution of ganglion cells in the retina of the three-toed sloth (Bradypus variegatus).

1. The distribution and size of retinal ganglion cells labelled with horseradish peroxidase (HRP) were studied in flat-mounted retinas of three-toed sloths. 2. Massive injections of HRP solution were made throughout the thalamus and midbrain in anesthetized sloths in order to retrogradely label the retinal ganglion cell population. Twenty to thirty h later the eyes were excised and the retinas flat-mounted and reacted with phenylenediamine-HCl and H2O2 to label ganglion cells, thus distinguishing them from other cells in the same retinal layer. 3. Ganglion cell density gradually increased from about 500 cells/mm2 at the far periphery to a peak of about 1,500 cells/mm2 in an area, termed the area centralis, deep in the inferior temporal retina. The presence of a vertical visual streak was also noted. 4. The area centralis contained a higher frequency of small ganglion cells than the peripheral retina where large cells predominated. 5. The unusual position of the area centralis and visual streak in the retina can be explained by the sloth's unique ability to rotate its head 180 degrees while climbing upside-down along horizontal branches so that the head is right-side-up. If it is assumed that the branch directly above the sloth's head needs to be visualized for accurate claw placement then the branch would be imaged on the inferior temporal retina in an area corresponding to the maximum density region.

Unusual distribution of ganglion cells in the retina of the three-toed sloth (Bradypus variegatus)

1. The distribution ans size of retinal ganglion cells labelled with horseradish preoxidase (HRP) were studied in flat-mounted retinas of three-toed sloths. 2. Massive injections of HRP solution were made throughout the thalamus and midbrain in anesthetized sloths in order to retrogradely label the retinal ganglion cell population. Twenty to thirty h later the eyes were and the retinas flat-mounted and reacted with phenylenediamine-HCL and H2O2 to label ganglion cells,thus distinguishing then from other cells int he same retinal layer. 3. Ganglion cell density graually increased from about 500 cell/mm2 at the far periphery to a peak of about 1,500 cells/mm2 in an area, termed the area centralis, deep in the inferior temporal retina. The presence of a vertical visual steak was also noted. 4. The area centralis contained a higher frequency of small ganglion cells than the peripheral retina where large cells preominated. 5. /the unusual postion of the area centralis and cisual streak in the retina can be explained by the slot's unique ability to rotate its head 180- while climbing upside-dow along horizontal branches so that the head is right-side-up. If it is assumed that the branch directly above the sloth's head needs to be visualized for accurate claw placement then the branch would be imaged on the inferior temporal retina in an area corresponding to the maximum density region

Pattern and extent of demyelination in the optic nerves of mice infected with Semliki Forest virus and the possibility of axonal sprouting.

The optic nerves of all mice infected with Semliki Forest virus (SFV) showed patchy demyelination, although the extent of demyelination varied between animals. There was a significant increase in small diameter unmyelinated fibres in the optic nerves of SFV infected mice, paralleled by a loss of small myelinated fibres, suggesting that the smaller myelinated fibres may be more susceptible to virus-induced demyelination. The small unmyelinated fibres were more numerous in the peripheral region of the optic nerve trunk, where blood vessels are found in greater numbers. In addition, a particularly large increase in the proportion of unmyelinated fibres which had diameters of less than 0.2 microns suggested the possibility of axonal sprouting.

Increase in axonal transport in demyelinating optic nerve fibres in the mouse infected with Semliki Forest virus.

Since the integrity of myelin and axon is closely linked with the transport of substances along the axon, changes in the fast and slow axonal transport in the demyelinating optic nerves of mice infected with Semliki Forest virus were studied. Radioactive analysis of the superior colliculi and the optic nerves was thus made following different survival times after an intravitreal injection of tritiated proline by autoradiographic and liquid scintillation counting techniques. The amounts of both the fast and slow axonal transport of proteins were significantly higher in the optic nerves of mice infected with the virus than those found in the optic nerves of control mice. These results suggest that demyelinating fibres send a signal to the perikarya which then responds by synthesizing more proteins.