A Study of the Development of Endotoxin-induced Inflammation in the Bovine Teat

Endotoxin-induced local inflammation was studied by frequent samplings in a bovine teat cistern model, which provides a unique possibility for in vivo studies of reactions in the teat without interference from the mammary gland. A rapid inflammatory response of rather short duration was elicited after endotoxin administration. An initial increase in the concentrations of bovine serum albumin and N-acetyl-β-D-glucosaminidase, indicating a disturbance in the epithelial integrity, was observed between 1 and 1.5 h post infusion (p.i.). Approximately 0.5 h later, the first influx of leukocytes, mainly neutrophils, appeared. The neutrophils tended to enter the teat cistern in several peaks occurring between 2.5 and 5 h p.i.. The sampling procedure decreased the accumulation of cells by approximately 40%, which was probably due to the removal of inflammatory mediators at an early stage. The parallel use of 2 teats instead of 1 had no major influence on the inflammatory process. This teat cistern model and the experimental procedure used should be suitable for further studies of the development of local inflammation.     

The Effect of Post Milking Teat Dip and Suckling on Teat Skin Condition,Bacterial Colonisation,and Udder Health

The teat skin of cows was scored (1: smooth as silk; 2: smooth; 3: slightly rough; 4: rough; 5: cracked; and 6: sores) by trained technicians who moved their fingers down the barrel of the teat with a light touch. Technicians ranked the same population of teats in the same rank order, but their mean values differed by half a score which probably could be related to the skin condition of their own fingers. Half udder experiments were carried out for 6 months at 4 farms with 35 to 52 cows each. A postmilking teat spray with 10% glycerol improved teat skin condition (p<0.10) compared with no teat spray. A postmilking teat spray with 120 ppm chlorine dioxide did not influence teat skin condition compared with no teat spray. No differences in udder health could be proven between treatments. Control studies revealed that 10% glycerol as an emollient of a postmilking teat spray improved teat skin condition within 3 weeks from being slightly rough to being smooth for lactating cows (p<0.05) but not for dry cows having smooth teat skin. Neither glycerol nor chlorine dioxide influenced absolute number of bacteria on teat skin after a challenge with Streptococcus uberis and Staphylococcus aureus but half life of S. aureus on unsprayed teats was longest (p = 0.05). Suckling made teat skin more rough than machine milking. Nevertheless, suckling lowered the number of esculin positive bacteria on the teat skin. We concluded that the condition of healthy teat skin (scores 1–4) has no influence on bacterial colonisation in the absence of cracks and sores (scores 5–6).     

Studies of Defence Mechanisms and Inflammatory Reactions in the Bovine Teat using a New Experimental Method

Bovine intramammary infections are usually caused by microorganisms entering through the teat canal. The teat canal normally acts as a mechanical and physiological barrier preventing bacteria from entering the teat cistern. This barrier can be broken, e.g. if the teat end is damaged, making it possible for bacteria to invade the teat cistern. If an infection is established inflammatory reac-tions will occur in the udder.     

Informative Value of Changes in Hand Skin Temperature in Response to Hypoxic Exposure

Residents of Magadan oblast with no experience of regular exposure to low ambient temperatures were examined to study the changes in hand skin temperature (HST) as a response to hypoxia. It was found that vasodilative responses followed by an increase in HST were typical of subjects demonstrating a high resistance to hypoxia during a 3-min rebreathing test and when breathing a gas mixture containing 8% O₂ and 92% N₂ for 14 min. Vasoconstrictor responses and a decrease in HST prevailed in subjects with a low resistance to hypoxia. Hand skin areas differed in the informative value of temperature changes in response to standard hypoxic exposure. Their characteristics may be used to select subjects for work under extreme environmental conditions and as an individual and specific marker of a human ecological portrait.__________Translated from Fiziologiya Cheloveka, Vol. 31, No. 3, 2005, pp. 108–117.Original Russian Text Copyright © 2005 by Maksimov.     

Temperature Regulation of Anthocyanin Accumulation in Apple Skin

The regulation of anthocyanin accumulation in apple skin (cv.Jonathan) by temperature was studied. In the field the increasein anthocyanin in the skin before harvest coincided with decreasingtemperatures and with increasing ethylene production by thefruit. In detached apples held under continuous white light,the optimum temperatures for anthocyanin accumulation were 12°C in unripe apples and 16–24 °C in ripe apples.These effects were explained by corresponding changes in thelevel of phenylalanine ammonia-lyase (PAL), a key enzyme offlavonoid synthesis. PAL levels were higher at low than at hightemperatures and higher in ripe than in unripe apples. In intermittentlight the effects of temperature were similar but levels ofPAL and anthocyanin were lower, particularly in unripe apples.It is concluded that temperature, in conjunction with ripeningand light, is an important factor regulating anthocyanin accumulationand that its effects are mediated by effects on the level ofPAL activity. Key words: Apple, Anthocyanin, Phenylalanine ammonia-lyase     

牛卵母细胞孤雌激活及马-牛异质克隆胚构建

首先用不同的激活剂孤雌激活体外成熟培养的牛卵母细胞,经试验获得:离子霉素、A23187和7%乙醇联合6-DMAP可有效地激活牛卵母细胞,并支持其发育到囊胚,但离子霉素激活效率显著优于其他两种(P<0.05);以10?S SOFaa 颗粒细胞为发育体系培养激活的成熟牛卵母细胞可得到较高的卵裂率和囊胚率(72.30%,14.91%)。其次,通过体外培养成年马皮肤成纤维细胞,将获得的成纤维细胞经血清饥饿培养后,作为核供体移入去核牛卵母细胞透明带下,电融合后,能得到融合的马牛重构胚,在交流电脉冲起始电压20V,持续时间10s,频率0.2MHz,结束电压15V,2次脉冲和融合间隔为0.125s的条件下,当融合电压为2.0kV/cm,脉冲时程为40μs时,重组胚的融合率和卵裂率最高(52.27%,71.74%)。     

Chronological Changes of Bovine Follicular Oocyte Maturation in Vitro

Chronological changes of bovine follicular cumulus-oocyte-complexesi were studied after in vitro maturation over a period of 48 h. According to their thickness and compactness of cumulus investments they were classified into 4 groups and cultured in enriched Ham’s F-10 medium with or without human chorionic gonadotrophin (hCG) and estradiolbenzoate (EB) for 0, 6, 12, 18, 21, 24, 27, 30 and 48 h. Representative samples were taken at each time interval for evaluation of nuclear maturation stages, ooplasm quality and size of the peri vitelline space (PVS). The results showed that oocyte nuclear breakdown (ONBD) required 6 to 12 h culture, and the peak of the first polar abstriction occurred at 24 h. The culture period required for ONBD and abstraction of the first polar body were related to the thickness and compactness of cumulus investments with and approximately 6 h delay in heavily compacted complexes. Ooplasm quality evaluation failed to show a clear trend, but the PVS increased in size from 0 h to 30 h and then, retracted again from 30 to 48 h. The overall maturation rate in the presence of hCG and EB was 79.1 %, and a substantial proportion (68.8 %) of nude or partially covered oocytes reached metaphase II stage. In the presence of hCG and EB no block at either metaphase I or at anaphase-telophase I was observed. In the absence of hCG and EB the percentage of oocytes reaching metaphase II was much lower (48.6%) in comparison with oocytes matured in the presence of these hormones (79.1 %). It was concluded a very high proportion of slaughterhouse oocytes could be matured in vitro and that the cumulus investments and addition of certain hormones affected the maturation rate.     

Electrocutaneous Sensitivity: Effects of Skin Temperature

The effect of human skin temperature on electrocutaneous sensitivity was examined using brief capacitive discharges. Stimuli were designed to ensure that sensory effects would be independent of skin resistance and would reflect underlying neural excitability as closely as possible. Skin temperature was manipulated by immersing the forearm in circulating hot or cold air. Detection thresholds on the arm and fingertip were raised by cooling, but were not altered by heating. Temperature-related sensitivity shifts were described by the same multiplicative factors for both threshold and suprathreshold levels. The temperature coefficient (Q₁₀) for cutaneous sensitivity under these conditions was approximately 1.3.     

Response Surface Methodology for Optimizing the Bovine Serum Albumin Fibrillation

Amyloid fibrils are considered as nanostructures that could be formed by ordered self-assembly of the partially-folded states of many different peptides or proteins. In this study, bovine serum albumin was used as a model protein whose ordered aggregation (fibrillation) was optimized. Response surface methodology (RSM) was used in a design that contained a total of 30 experimental trials. The first 24 were organized in a factorial design and from 25 to 30 involved the replications of the central points. Data obtained from RSM were subjected to the analysis of variance (ANOVA) and analyzed using a second order polynomial equation. Subsequent testing of the suggested experimental parameters was done in vitro with Congo red spectrophotometric assay. Protein concentration, pH, temperature and time of incubation were the variables used in this study. Responses were assessed by measuring absorbance in 540?nm (characteristic of amyloid formation) and maximal wavelength. Concomitant effects of variables were assessed in surface plots that each considered two of the variables. Interestingly, the pattern obtained by monitoring absorbance at 540?nm and absorbance in maximal wavelength were identical in most cases. We are reporting the optimum concentration of protein, pH, temperature and time at 5?mg?ml^?1, 3.02 and 72?°C and 48?h, respectively. Our findings suggest that use of Congo red spectrophotometric test, as a simple and affordable assay could be suggested as a first test for assessing fibril formation of proteins. Absorbance in maximal wavelength is recommended as a significant indication of fibril formation.     

A Method to Record the Surface Temperature Distribution of the Skin as a Safe and Ergonomic Approach to Diagnosis and Monitoring in Breast Cancer

Biophysics - Abstract—This article discusses the relevant results on early and safe detection of breast cancer with a thermal volume tomography (TVT) device produced in Russia. The device...     

Phase Transition Temperature and Chilling Sensitivity of Bovine Oocytes

A limiting factor for achieving cryopreservation of oocytes is direct chilling injury (DCI), which occurs during cooling. DCI, or cold shock, is defined as an irreversible damage expressed shortly after exposure to low, but not freezing, temperatures. The primary target of DCI is thought to be the plasma membrane. Recently, an association between DCI in sperm and the thermotropic phase transition of their membrane lipids was demonstrated. In the present study, we examined the phase transition of the membrane lipids of immature andin vitro-matured bovine oocytes during cooling, using Fourier transform infrared spectroscopy (FTIR). The phase transition of the membrane lipids of oocytes at the germinal vesicle (GV) stage occurred between 13 and 20°C, while a very broad phase transition, which centered around 10°C, was observed for mature oocytes (MII) stage. Thermotropic phase transitions were demonstrated to be related to the temperature at which DCI affected the integrity of the oocyte membranes. When immature oocytes were cooled to 13°C, fewer oocytes (40%) retained their membrane integrity than after exposure to 4°C (51%) or holding them at 38°C (78%), (as determined by the Fluorescein Diacetate-FDA test). This finding might suggest that holding immature oocytes at the phase transition temperature is more damaging to their membranes than exposure to lower temperatures. By contrast, no significant differences in membrane integrity were observed whenin vitro-matured oocytes were cooled to the same temperatures. Subsequently, GV oocytes were cooled to 4°C, and 26% underwent maturation and 19% underwent fertilizationin vitro. In vitro-matured oocytes that were cooled to 4°C displayed a slightly decreased rate of fertilization; the overall fertilization was 60% with 24% polyspermy, rather than the 76% fertilization rate with 12% polyspermy obtained with those not subjected to cooling. The high rate of polyspermy indicates that a site(s) other than the plasma membrane is affected during cooling of bovine oocytes. Nucleated bovine GV oocytes were electrofused within vitro-matured and enucleated oocytes, and then cooled to 4°C. Evaluation of the membrane integrity of the fused oocytes showed that these oocytes are chilling resistant, which strongly suggests that alteration of the membrane composition of an oocyte can change the cell's susceptibility to low temperatures. This finding led to an improvement in the survival of oocytes after cryopreservation.