Watanabe Applied Technology (Brazil)

To describe clinical uses of the class of alpha-2 receptor agonist medications in the treatment of traumatic brain injury sequelae. Clinicians treating patients with traumatic brain injuries (TBI) will often see patients who have been prescribed alpha-2 agonists or may consider initiating use. Narrative review. This class of medication has a number of different indications (and side effects), and drugs within this class also have some unique properties that can be important factors in clinical decision-making. Although this class of medications has been available for many years, there is still much emerging information that is pertinent for the treatment of patients with TBI.

The present study evaluated the effects of melatonin (MEL) during IVM of bovine cumulus-oocyte complexes (COC). The COC were cultured in droplets (25–30/100 µL) under mineral oil at 38.5°C and 5% CO2 in air. Medium (TCM199 + 0.1% polyvinyl alcohol, 0.25 mM sodium pyruvate, and 25 µg mL–1 of gentamycin) was supplemented with FSH (0.5 µg mL–1), MEL (10–9 and 10–6 M), or no hormones (control). In experiment 1, oocytes were assessed for nuclear maturation rates (6, 12, 18, and 24 h of IVM). In experiment 2, relative abundance of antioxidant enzymes copper, zinc superoxide dismutases (CuZnSOD), manganese superoxide dismutases (MnSOD), and glutathione peroxidase (GPx) was evaluated in oocytes and cumulus cells (0 and 24 h) by real time PCR. The immature group was the reference and endogenous controls were actin β and glyceraldehyde 3-phosphate dehydrogenase. In experiment 3, nuclear fragmentation in cumulus was assessed by TUNEL and flow cytometry (24 h). In experiment 4, embryo development after in vitro fertilization and culture was analysed (cleavage rates D2, blastocysts Day 8, and hatching Day 10). The control group was matured in complete IVM medium (10% FCS, 0.5 µg mL–1 of FSH, 50 µg mL–1 of LH, and 1 µg mL–1 of oestradiol). The MEL and FSH groups were the same. Data (3–4 replicates) were analysed by Chi-square (experiment 1; GraphPad Prism) or ANOVA and Tukey test (SAS, 1995) and 5% significance. At 6 h of IVM, all oocytes were at germinal vesicle stage. At 12 h, hormone groups had similar metaphase I rates (71–81%, P > 0.05) and were superior to control (54%, P < 0.05). At 18 h, metaphase II (MII) rates were similar (57–74%, P > 0.05). After 24 h, MEL (10–9 and 10–6 M) was similar to control (51, 67, and 69% MII, respectively, P > 0.05) and FSH had the highest MII rates (90%, P > 0.05). The 10–6 M MEL was similar to FSH (P > 0.05). Antioxidant enzymes were unaffected in oocytes (P > 0.05%). The CuZnSOD transcripts increased in cumulus after IVM with 10–6 M MEL compared to immature cells (3.5 and 1.0, respectively, P < 0.05). Control, FSH, and 10–9 M MEL did not differ (2.1–2.5, P > 0.05) or increased relative to immature cells (P < 0.05), but were inferior to 10–6 M MEL (P < 0.05). The MnSOD relative abundance was similar for immature, control, and 10–9 M MEL (1.0–3.0, P > 0.05). The 10–6 M MEL increased MnSOD relative to immature cells (3.0 and 1.0, respectively, P < 0.05), but was similar to control and 10–9 M MEL (P > 0.05). The FSH showed the highest levels (9.1, P < 0.05); GPx4 transcripts were not affected (P > 0.05). Nuclear fragmentation in cumulus was not influenced (33.4–41.5/10 000 cells; P > 0.05). Embryo development rates were similar for all groups (cleavage: 82–87%, blastocysts: 49–54, hatching: 91–96%, P > 0.05). In conclusion, MEL during IVM stimulates meiosis resumption at rates similar to FSH and embryo development similar to FSH and complete IVM medium. The MEL increased CuZnSOD expression in cumulus, but no parallel effect was observed on nuclear fragmentation.

Human embryonic stem cells (ESC) lines to be used for cell therapies must be created and maintained under strict conditions, excluding the use of undefined supplements. Two key steps in the creation of a new embryonic stem cell line are adherence to the substrate and derivation towards the formation of a primary colony. The bovine parthenote embryo model was used to test different matrices of gelatin nanofibers and gelatin/galactomannan films to be used for ESC derivation and culturing. Gelatin/galactomannan films were made in two concentrations of galactomannan, 0.1 and 0.3%, in an aqueous solution of gelatin and tested for gel cytotoxicity using cumulus cells (CCs). CCs showed normal cell morphology, with no sign of lysis or degeneration in any of the matrices tested. Inner cell masses of parthenote blastocysts (n=116) were placed onto the gel matrices for culture. There were three or four repeats for each matrix. Our results showed a good rate of inner cell mass (ICM) adherence on the gelatin/galactomannan films (41%–44%) and one derivative of the gel nanofiber (17% adherence to the substrate). These results encouraged us to try new gelatin formulations to increase the rates of derivation and cell proliferation under defined culture conditions to comply with good manufacturing practice directives for the potential therapeutic use of ESCs.

Oocytes from dairy cattle and buffaloes have severely compromised developmental competence during summer. While analysis of gene expression is a powerful technique for understanding the factors affecting developmental hindrance in oocytes, analysis by real-time reverse transcription PCR (RT-PCR) relies on the correct normalization by reference genes showing stable expression. Furthermore, several studies have found that genes commonly used as reference standards do not behave as expected depending on cell type and experimental design. Hence, it is recommended to evaluate expression stability of candidate reference genes for a specific experimental condition before employing them as internal controls. In acknowledgment of the importance of seasonal effects on oocyte gene expression, the aim of this study was to evaluate the stability of expression levels of ten well-known reference genes (ACTB, GAPDH, GUSB, HIST1H2AG, HPRT1, PPIA, RPL15, SDHA, TBP and YWHAZ) using oocytes collected from different categories of dairy cattle and buffaloes during winter and summer. A normalization factor was provided for cattle (RPL15, PPIA and GUSB) and buffaloes (YWHAZ, GUSB and GAPDH) based on the expression of the three most stable reference genes in each species. Normalization of non-reference target genes by these reference genes was shown to be considerably different from normalization by less stable reference genes, further highlighting the need for careful selection of internal controls. Therefore, due to the high variability of reference genes among experimental groups, we conclude that data normalized by internal controls can be misleading and should be compared to not normalized data or to data normalized by an external control in order to better interpret the biological relevance of gene expression analysis.

The system and performance of the developed data processing unit that detects and classifies the fabric defects as compared with the corresponding data of the normal region by using the statistical processing are studied.The intensity distribution are sampled from the electro-optical output by scanning and the mean value of integration in each scanning times expressing the characters of the fabric without defects are obtained by the pre-processing part. Then the several parameters needed for detecting the defects are calculated according to these data by the digital computer.The threshold values to separate the abnormal regions from normal are decided by way of K times of the standard deviation obtained from the histograms of the electro-optical output by the reflection or transparency from fabrics, also the values of K are judged by many experimental results.The defect informations expressing the existing positions and sizes according to the threshold logic are detected by the analogue comparators and the peak holding circuits. These informations are changed into the binary coded signals by A-D converters and fed to the digital computer.The digital computer prints out the several data fed from the data processing unit and the results calculated by these data.

In scanning the surface of fabrics with a light spot, we can get the power of the reflection or transparency that periodically changes according to disposition, diameter or bend of the threads. Then, on application of a digital computer, a digital pattern of fabric is represented by a twodimentional array of points showing the reflection and transparency distributions of fabrics by a suitable scanning light spot.On detecting the defects by the reflection or transparency from fabrics, it is necessary to design the size and shape of the scanning light spot at the fabrics in consideration of the some kinds of defects and the inherent noises being due to the normal fabric structures, also the frequency band determined by the resolving power and the scanning speed that are suitable to the electric amplifier system.In general, the spaces between the threads in fabrics are much smaller than the thread diameters. When width of the scanning light spot is smaller than the thread diameter, we can detect the characters of the thread and thread interval in every each warp direction. When length of the scanning light spot is a little larger than the thread diameter, average characters of a few threads will be detected, eliminating the noises caused by the twists and fluffes of the threads. In our experiments, the width of the scanning light spot was set 0.2mm and length 5mm in consideration of practrical usefulness.Moreover, the frequency band f needed to design of the electro-optical amplifier is given by f=V/2b where V is scanning speed of the light spot, 240×103mm/sec, and b is resolving power, 0.2mm.Confirmation of the size and shape of the light spot, transient properties and frequency responses of the signal transduser was discussed with the some experimental equipments.