Aspirin Enhances the Protection of Hsp90 from Heat-Stressed Injury in Cardiac Microvascular Endothelial Cells Through PI3K-Akt and PKM2 Pathways.

Heat stress (HS) often causes sudden death of humans and animals due to heart failure, mainly resulting from the contraction of cardiac microvasculature followed by myocardial ischemia. Cardiac microvascular endothelial cells (CMVECs) play an important role in maintaining vasodilatation. Aspirin (ASA) is well known for its protective abilities of febrile animals. However, there is little knowledge about molecular resistance mechanisms of CMVECs and which role ASA may play in this context. Therefore, we used a heat stress model of rat cardiac microvascular endothelial cell cultures in vitro and investigated the cell injuries and molecular resistance mechanism of CMVECs caused by heat stress, and the effect of aspirin (ASA) on it. HS induced severe pathological damage of CMVECs and cellular oxidative stress and dysfunction of NO release. Hsp90 was proven to be indispensable for resisting HS-injury of CMVECs through PI3K-Akt and PKM2 signaling pathways. Meanwhile, PKM2 functioned in reducing Akt phosphorylation. ASA treatment of CMVECs induced a significant expression of Hsp90, which promoted both Akt and PKM2 signals, which are beneficial for relieving HS damage and maintaining the function of CMVECs. Akt activation also promoted HSF-1 that regulates the expression of Hsp70, which is known to assist Hsp90's molecular chaperone function and when released to the extracellular liquid to protect myocardial cells from HS damage. To the best of our knowledge, this is the first study to show that HS damages CMVECs and the protection mechanism of Hsp90 on it, and that ASA provides a new potential strategy for regulating cardiac microcirculation preventing HS-induced heart failure.

Do Pigs Have Adequate Space in Animal Transportation Vehicles?-Planimetric Measurement of the Floor Area Covered by Finishing Pigs in Various Body Positions.

In this study, the floor area covered by individual finishing pigs in various body positions was measured using a contrast-based planimetric method for computer-assisted analysis of two-dimensional images. Two hundred and thirty-two finishing pigs were weighed during the last fifth of the fattening period and measured in different body positions using contrast-based planimetry. Thirteen body positions were defined based on characteristic directions of the head, legs and body. The lowest average covered floor area was found for body position A (pig standing up straight, nose touching the ground) with 0.288 ± 0.026 m2. The highest average covered floor area for a standing pig amounted to 0.335 ± 0.030 m2 in body posture ES (pig standing curved sideways, head raised above the dorsal line) and, for a lying pig, 0.486 ± 0.040 m2 (posture LL, pig lying in fully lateral recumbent position). The covered floor surface significantly depended on the weight of the animal and the body posture. Allometric estimations previously described for calculating the floor area physically covered by a pig's body are not consistently precise in depicting the actual areas covered. The minimal floor area offered in animal transportation vehicles, according to European legislation, is insufficient in the case of all pigs lying in the fully recumbent position simultaneously, without the pigs being forced to partially overlap one another. Therefore, both allometric formulas and legislation should be modified on the basis of these results and further studies with pigs of modern genetic origin should be conducted.

Expression and location of HSP60 and HSP10 in the heart tissue of heat-stressed rats.

The present study aimed to analyze the expression levels and localizations of heat shock protein (HSP) 60 and HSP10 in the heart tissue of rats subjected to heat stress (42°C) for 0, 20, 80 and 100 min. Histopathological injuries and increased serum activities of serum lactate dehydrogenase and creatine kinase isoenzyme MB were detected in the heated rat myocardial cells. These results suggested that heat stress-induced acute degeneration may be sufficient to cause sudden death in animals by disrupting the function and permeability of the myocardial cell membrane. In addition, the expression levels of HSP60 were significantly increased following 20 min heat stress, whereas the expression levels of its cofactor HSP10 were not. Furthermore, the location of HSP60, but not of HSP10, was significantly altered during periods of heat stress. These results suggested that HSP60 in myocardial tissue may be more susceptive to the effects of heat stress as compared with HSP10, and that HSP10 is constitutively expressed in the heart of rats. The expression levels and localizations of HSP60 and HSP10 at the different time points of heat stress were not similar, which suggested that HSP60 and HSP10 may not form a complex in the heart tissue of heat-stressed rats.

Antimicrobial-Resistant Escherichia coli Survived in Dust Samples for More than 20 Years.

In a retrospective study, 119 sedimentation dust samples stored between five and 35 years from various barns of intensive livestock farming were evaluated for the occurrence of cultivatable Escherichia coli. Growth of E. coli occurred in 54 samples. Successful cultivation was achieved in samples from as early as 1994. The frequency of detection increased from earlier to later time periods, but the concentrations, which ranged between 3.4 × 10(2) and 1.1 × 10(5) colony-forming units per gram, did not correlate with sample age (Spearman rank correlation; p > 0.05). We hypothesize that E. coli cells survived in dust samples without cell division because of the storage conditions. Dry material (dust) with low water activities (arithmetic mean < 0.6) and storage at 4°C in the dark likely facilitated long-term survival. E. coli were isolated on MacConkey agar with and without ciprofloxacin supplementation. For 110 isolates (79 from non-supplemented media and 31 from supplemented media), we determined the E. coli phylotype and antimicrobial resistance. Six phylogenetic groups were identified. Phylogroups A and B1 predominated. Compared to group A, phylogroup B1 was significantly associated with growth on ciprofloxacin-supplemented media (chi-square test, p = 0.003). Furthermore, the antibiotic resistance profiles determined by a microdilution method revealed that isolates were phenotypically resistant to at least one antimicrobial substance and that more than 50% were resistant to a minimum of five out of 10 antibiotics tested. A linear mixed model was used to identify factors associated with the number of phenotypic resistances of individual isolates. Younger isolates and isolates from fattening poultry barns tended to be resistant to significantly more antibiotics than older isolates and those from laying-hen houses (p = 0.01 and p = 0.02, respectively). Sample origin and storage conditions may have influenced the number of antimicrobial resistances. Overall, we found that under particular conditions, dust from farm animal houses can be reservoirs for antimicrobial-resistant E. coli for at least 20 years. The survival strategies that allow E. coli to survive such long periods in environmental samples are not fully understood and could be an interesting research topic for future studies.

Localization and expression of heat shock protein 70 with rat myocardial cell damage induced by heat stress in vitro and in vivo.

The aim of the present study was to investigate the association between heat shock protein (Hsp) 70 expression kinetics and heat stress­induced damage to rat myocardial cells in vitro and in vivo. The results showed that the activity of heart injury­associated enzymes, including aspartate aminotransferase and creatine kinase, significantly increased and myocardial cells developed acute histopathological lesions; this therefore suggested that heat stress altered the integrity of myocardial cells in vitro and in vivo. Levels of Hsp70 in vitro decreased following the initiation of heat stress and then steadily increased until heat stress ceased at 100 min; however, in vivo studies demonstrated a gradual increase in Hsp70 levels in the heart cells of rats from the initiation of heat stress, followed by a sharp decline at 100 min. These results indicated that the cells sustained different degrees of injury in vivo compared with those sustained in vitro, this may be due to different regulatory mechanisms in the two environments. Intracytoplasmic Hsp70 signaling was significantly reduced at 60 min in vitro, compared with that of the in vivo study, indicating that Hsp70 consumption may have exceeded its production prior to 60 min of heat stress, and following 60 min the cells produced sufficient Hsp70 protein for their protection against heat stress. Hsp70­positive signals in the cytoplasm of heart cells in vivo were more prominent in the intact areas compared with those of the degenerated areas and the density of Hsp70­positive signals was significantly reduced following 60 min of heat stress. In conclusion, comprehensive comparisons of enzymes, cell morphology and Hsp70 levels indicated that decreased levels of Hsp70 were associated with the reduced protective effect on myocardial cells in vitro and in vivo.

Comparative analysis of αB-crystallin expression in heat-stressed myocardial cells in vivo and in vitro.

Relationships between αB-crystallin expression patterns and pathological changes of myocardial cells after heat stress were examined in vitro and in vivo in this study using the H9C2 cell line and Sprague-Dawley rats, respectively. Histopathological lesions, characterized by acute degeneration, karyopyknosis and loss of a defined nucleus, became more severe in rat hearts over the course of heat stress treatment from 20 min to 100 min. The expression of αB-crystallin in rat hearts showed a significant decrease (P<0.05) throughout the heat stress treatment period, except at the 40 min time point. Likewise, decreased αB-crystallin expression was also observed in the H9C2 cell line exposed to a high temperature in vitro, although its expression recovered to normal levels at later time points (80 and 100 min) and the cellular damage was less severe. The results suggest that αB-crystallin is mobilized early after exposure to a high temperature to interact with damaged proteins but that the myocardial cells cannot produce sufficient αB-crystallin for protection against heat stress. Lower αB-crystallin expression levels were accompanied by obvious cell/tissue damage, suggesting that the abundance of this protein is associated with protective effects in myocardial cells in vitro and in vivo. Thus, αB-crystallin is a potential biomarker of heat stress.

Cognitive enrichment in piglet rearing: an approach to enhance animal welfare and to reduce aggressive behaviour.

It is known that pigs raised in enriched environments express less aggressive behaviour. For this reason, a new method of cognitive environmental enrichment was experimented at the University of Veterinary Medicine Hannover, Germany. In the first phase, 78 suckling piglets were trained to learn the link between a sound given by an electronic feeder and a feed reward in the form of chocolate candies during a period of 8 days. In the second phase, the same piglets were used in resident-intruder tests to verify the potential of the feeding system to interrupt aggressive behaviour. The analysis of all training rounds revealed that piglets learned the commands during 8 days of training and the interest of the piglets increased within training days (P < 0.05). In the resident-intruder test, 79.5% of aggressive interactions were broken by feeder activation. In interactions where either the aggressor or the receiver reacted, a high number of fights were stopped (96.7% versus 93.1%) indicating that it was not relevant if the aggressor or the receiver responded to the feeder activation. We conclude that the electronic feeding system has the potential to be used as cognitive enrichment for piglets, being suitable for reducing aggressive behaviour in resident-intruder situations.

Isoflurane-anaesthesia used for piglet-castration: a bacteriological assessment of the anaesthetic device.

Castration of piglets with the use of isoflurane-anaesthesia in combination with an analgesic has been proven to be a welfare-friendly approach. However, castration is performed with an equipped anaesthetic device which is not profitable for small farms. Thus, this study aimed at investigating whether sharing the anaesthetic device among farms results in an elevated risk of bacteriological contamination and further spreading in spite of a thorough disinfection. The study included two groups of organic farms. Piglets (n = 1579) were anaesthetised with isoflurane supplied from an equipped device and castrated. Stationary anaesthetic devices were used in the first group of farms, whereas farms in the second group shared one device. Each farm was visited four times and the colony forming units (CFU) of total mesophilic bacteria, Staphylococcus spp., Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli were determined. Sampling took place before castration and after disinfection, and included snout masks, retaining fixtures, pedals and wheels of the device (n = 376). The results indicated presence of Staphylococcus species in 56.5% and 40.3% of samples obtained from farms using stationary and shared devices, respectively. MRSA was detected in 2.4% of the samples and only one pathogenic Shiga toxin-producing E. coli (Stx2e, F18) could be detected. Bacterial counts were lower on the shared device than on the stationary devices (p = 0.038), particularly on restraining fixtures (p < 0.05). In both groups wheels were the most and pedals the least contaminated materials (p < 0.05). It is concluded that sharing an anaesthetic device on several farms does not increase bacteriological contamination after a thorough disinfection and thus imposing no additional hygiene risk to farms with similar hygienic conditions.

Localization and expression of Hsp27 and αB-crystallin in rat primary myocardial cells during heat stress in vitro.

Neonatal rat primary myocardial cells were subjected to heat stress in vitro, as a model for investigating the distribution and expression of Hsp27 and αB-crystallin. After exposure to heat stress at 42°C for different durations, the activities of enzymes expressed during cell damage increased in the supernatant of the heat-stressed myocardial cells from 10 min, and the pathological lesions were characterized by karyopyknosis and acute degeneration. Thus, cell damage was induced at the onset of heat stress. Immunofluorescence analysis showed stronger positive signals for both Hsp27 and αB-crystallin from 10 min to 240 min of exposure compared to the control cells. According to the Western blotting results, during the 480 min of heat stress, no significant variation was found in Hsp27 and αB-crystallin expression; however, significant differences were found in the induction of their corresponding mRNAs. The expression of these small heat shock proteins (sHsps) was probably delayed or overtaxed due to the rapid consumption of sHsps in myocardial cells at the onset of heat stress. Our findings indicate that Hsp27 and αB-crystallin do play a role in the response of cardiac cells to heat stress, but the details of their function remain to be investigated.

Structure and possible functions of constant-frequency calls in Ariopsis seemanni (Osteichthyes, Ariidae).

In the 1970s, Tavolga conducted a series of experiments in which he found behavioral evidence that the vocalizations of the catfish species Ariopsis felis may play a role in a coarse form of echolocation. Based on his findings, he postulated a similar function for the calls of closely related catfish species. Here, we describe the physical characteristics of the predominant call-type of Ariopsis seemanni. In two behavioral experiments, we further explore whether A. seemanni uses these calls for acoustic obstacle detection by testing the hypothesis that the call-emission rate of individual fish should increase when subjects are confronted with novel objects, as it is known from other vertebrate species that use pulse-type signals to actively probe the environment. Audio-video monitoring of the fish under different obstacle conditions did not reveal a systematic increase in the number of emitted calls in the presence of novel objects or in dependence on the proximity between individual fish and different objects. These negative findings in combination with our current understanding of directional hearing in fishes (which is a prerequisite for acoustic obstacle detection) make it highly unlikely that A. seemanni uses its calls for acoustic obstacle detection. We argue that the calls are more likely to play a role in intra- or interspecific communication (e.g. in school formation or predator deterrence) and present results from a preliminary Y-maze experiment that are indicative for a positive phonotaxis of A. seemanni towards the calls of conspecifics.

Tail biting in pigs--causes and management intervention strategies to reduce the behavioural disorder. A review.

One of the largest animal welfare problems in modern pig production is tail biting. This abnormal behaviour compromises the well-being of the animals, can seriously impair animal health and can cause considerable economic losses. Tail biting has a multifactorial origin and occurs mainly in fattening pigs. High stocking densities, poor environment and bad air quality are seen as important factors. However, it is presumed that a plurality of internal and external motivators in intensive pig production can trigger this behaviour which is not reported in sounders of wild boars. The aim of this review is to summarize the causes and the effects of tail biting in pigs and present management strategies that are likely to reduce its incidence. In particular, management strategies by applying Precision Livestock Farming (PLF) technologies to monitor and control the behaviour of the pigs may be suitable to detect the outbreaks of tail biting at an early stage so that counter measures can be taken in time.

How do pigs behave before starting an aggressive interaction? Identification of typical body positions in the early stage of aggression using video labelling techniques.

The aim of this study was to identify, quantify, and describe pre-signs of aggression in pigs and the early stages of aggressive interactions. The experiment was carried out at a commercial farm on a group of 11 male pigs weighing on average 23 kg and kept in a pen of4 m x 2.5 m. In total 8 hours were videorecorded during the first 3 days after mixing. As a result, 177 aggressive interactions were identified and labelled to find pre-sign body positions before aggressive interactions, attack positions and aggressive acts performed from these positions. A total of 12 positions were classified as pre-signs (P1-P12) and 7 of them were identified immediately at the start of aggressive interactions (P6-P12). Most common pre-sign positions were P3-pigs approaching and facing each other (24%) and P2-initiator pigs approaching from the lateral side (18%). In 80% of the cases the duration of pre-signs was 1-2 sec 72% of all aggressive interactions were short (1 to 10 sec). The most frequent attack positions were P12-inverse parallel (39.5%), P7-nose to nose, 90 degrees (19.77%) and P9-nose to head (13.5%). The most frequent aggressive acts from attack positions were head knocking (34.4%), pressing (34.4%) and biting of different body parts (29.4%). Head knocking was mostly observed in relation to P7 and P2 positions and biting was common in the P7 position. In conclusion, pigs adopt specific pre-signs and body positions before the escalation of aggressive interactions. This could be used as potential sign to identify a beginning aggression.

[Reduction of livestock-associated methicillin-resistant staphylococcus aureus (LA-MRSA) in the exhaust air of two piggeries by a bio-trickling filter and a biological three-step air cleaning system]. / Abscheidung von "Livestock-associated" Methicillin-resistenten Staphylococcus aureus (LA-MRSA) aus der Abluft zweier Mastschweineställe mit einem Riesel- bettfilter und einer dreistufigen Abluftreinigungsanlage.

"Livestock-associated" Methicillin-resistent Staphylococcus aureus (LA-MRSA) are frequently found in the air of piggeries, are emitted into the ambient air of the piggeries and may also drift into residential areas or surrounding animal husbandries.. In order to reduce emissions from animal houses such as odour, gases and dust different biological air cleaning systems are commercially available. In this study the retention efficiencies for the culturable LA-MRSA of a bio-trickling filter and a combined three step system, both installed at two different piggeries, were investigated. Raw gas concentrations for LA-MRSA of 2.1 x 10(2) cfu/m3 (biotrickling filter) and 3.9 x 10(2) cfu/m3 (three step system) were found. The clean gas concentrations were in each case approximately one power of ten lower. Both systems were able to reduce the number of investigated bacteria in the air of piggeries on average about 90%. The investigated systems can contribute to protect nearby residents. However, considerable fluctuations of the emissions can occur.

The potential of a new air cleaner to reduce airborne microorganisms in pig house air: preliminary results.

There is a need for technical solutions to reduce the concentrations of bioaerosols in the air and in the exhaust air of livestock buildings. A prototype of an air washer combined with a UV-irradiation system was positioned in a commercial pig fattening unit to test its efficiency of reducing culturable airborne microorganisms. No significant reduction in airborne bacteria and fungi was observed when untreated air passed through the device. However, when the air washer or the UV-irradiation system was activated, the concentrations of mesophilic aerobic bacteria, methicillin resistant Staphylococcus aureus and mesophilic aerotolerant cocci were reduced significantly (p < 0.01). Washing the air reduced bacteria by 84 to 96% and the relative reduction due to UV-irradiation ranged between 55 and 90%. The highest relative reduction in airborne bacteria (90 to 99%) was detected when the air washer and the UV-irradiation systems were in simultaneous operation. The concentration of total airborne fungi was reduced significantly (p < 0.05) only when the air was washed and UV-irradiated. Although these preliminary results provided significant and comprehensible findings, long-term studies are required to assess the efficiency of the device in more detail.The combination of air washing and UV-irradiation seem to be a useful technique for abating airborne microorganisms within or emitting from piggery buildings. However, some technical problems remain, such as the deposition of particulate matter on the surface of UV-irradiators and the consumption of fresh water by the air washer. These issues must be resolved before the system may be implemented for general practice.

[Planimetric measurement of floor space covered by pullets]. / Planimetrische Ermittlung des Mindest- platzbedarfs von Junghennen.

The available floor space has a strong impact on the execution of various behaviours of laying hens. Presently, in Germany detailed requirements on the housing of pullets are insufficient. In order to get a first approximation, the floor space covered by pullets was determined by the colour contrast planimetric method KobaPlan. The measurements on standing and sitting pullets were done on a random sample of Lohmann Brown (LB), Lohmann Tradition (LT), Lohmann Selected Leghorn (LSL) and Dekalb White (DW) hybrids from the 6th week of life to 18/20 weeks at regular intervals. The hens were weighed and photographed digitally in a specific planimetric box (n = 2600 photographs from pullets in standing and 1360 in sitting position). Afterwards the KobaPlan software program calculated the animal area.The results showed a correlation between floor space covered by the pullets in standing position and the live weight (R2 = 0.80-0.96). The mean floor space covered by LB and LT at the end of rearing (body weight 1450 +/- 25 g LB and 1500 +/- 25 g LT) was 422.4 +/- 41.9 cm2 (LB) and 446.7 +/- 49.0 cm2 (LT) stand- ing, respectively 448.0 +/- 51.0 cm2 (LB) and 464.5 +/- 42.6 cm2 (LT) sitting. LSL and DW (body weight 1300 +/- 25 g) used 371.0 +/- 41.3 cm2 (LSL) and 349.3 +/- 26.3 cm2 (DW), respectively in standing and 379.5 +/- 41.2 cm2 (LSL) in sitting position. Maximum stocking density recommendation for pullets based on these planimetric results compared with the space allowance in alternative housing systems for laying hens are between eleven and 14 birds/m2. To verify this stocking density recommendation for pullets further studies should be complemented by specific ethological observations.

Hsp70 and HSF-1 expression is altered in the tissues of pigs transported for various periods of times.

The aim of this study was to assess changes of Hsp70 and HSF-1 protein and mRNA expression in stress-sensitive organs of pigs during transportation for various periods of time. Twenty pigs were randomly divided into four groups (0 h, 1 h, 2 h, and 4 h of transportation). A significant increased activity of AST and CK was observed after 1 h and 2 h of transportation. Histopathological changes in the heart, liver, and stomach indicated that these organs sustained different degrees of injury. Hsp70 protein expression in the heart and liver of transported pigs did not change significantly while it increased significantly (p < 0.05) in the stomach. Hsp70 mRNA levels decreased significantly (p < 0.05) in the heart after 4 h of transportation. However, mRNA expression increased significantly in the liver after 1 (p < 0.05) and 4 h (p < 0.01) of transportation, and increased significantly in the stomach of the transported pigs after 1, 4 (p < 0.01), and 2 h (p < 0.05). HSF-1 levels were reduced at 1 and 4 h (p < 0.05) only in the hearts of transported pigs. These results indicate that Hsp70 mediates distinct stress-related functions in different tissues during transportation.

Longitudinal study of the contamination of air and of soil surfaces in the vicinity of pig barns by livestock-associated methicillin-resistant Staphylococcus aureus.

During 1 year, samples were taken on 4 days, one sample in each season, from pigs, the floor, and the air inside pig barns and from the ambient air and soil at different distances outside six commercial livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA)-positive pig barns in the north and east of Germany. LA-MRSA was isolated from animals, floor, and air samples in the barn, showing a range of airborne LA-MRSA between 6 and 3,619 CFU/m(3) (median, 151 CFU/m(3)). Downwind of the barns, LA-MRSA was detected in low concentrations (11 to 14 CFU/m(3)) at distances of 50 and 150 m; all upwind air samples were negative. In contrast, LA-MRSA was found on soil surfaces at distances of 50, 150, and 300 m downwind from all barns, but no statistical differences could be observed between the proportions of positive soil surface samples at the three different distances. Upwind of the barns, positive soil surface samples were found only sporadically. Significantly more positive LA-MRSA samples were found in summer than in the other seasons both in air and soil samples upwind and downwind of the pig barns. spa typing was used to confirm the identity of LA-MRSA types found inside and outside the barns. The results show that there is regular airborne LA-MRSA transmission and deposition, which are strongly influenced by wind direction and season, of up to at least 300 m around positive pig barns. The described boot sampling method seems suitable to characterize the contamination of the vicinity of LA-MRSA-positive pig barns by the airborne route.

[Investigations into the use of respiratory masks for reducing the MRSA-exposure of veterinarians visiting regularly pig herds--first experiences]. / Zum Einsatz von Staubmasken zur Senkung der MRSA-Exposition von Tieräirzten in der Bestandsbetreuung von Schweinebeständen- eine Pilotstudie.

The study presents first experiences on the controlled use of respiratory masks against Methicillin-resistant Staphylococcus aureus (MRSA) in a multi-person veterinary pig practice. Seven veterinarians entered the trial (five wearing masks, two wearing no masks) after the veterinarians had performed a decolonisation protocol. The pig herds were visited regularly by the veterinarians during the study period. The five "trial" veterinarians wore gloves and respiratory masks for at least 30 days and 30 farm visits. The two "control" veterinarians wore gloves only. Nasal swabs were collected at a seven day interval. Swabs and ten masks per "trial" veterinarian were bacteriologically tested for MRSA including MLST- and spa-typing. The study showed a high MRSA-exposure for the veterinarians, since 68% of the masks were tested positive for MRSA. However, four vets stayed MRSA-negative while using the masks. Only one of the"trial" veterinarians became positive after two weeks. After the masks were not worn any more, two veterinarians returned to colonisation soon again. The two "control" veterinarians turned positive after 26 and 54 days, respectively. The high finding-rates of MRSA in the masks proof an enormous risk of nasal colonisation during routine work.The results of our study do not proof the potential of respiratory masks to prevent nasal colonisation of veterinarians with MRSA. However, there are no hints, that the proper use of masks could be a risk factor for becoming colonised. Further details of the proper use of masks and the quantification of their protective potential need further studies on a larger scale.

Variation in the expression of Hsp27, αB-crystallin mRNA and protein in heart and liver of pigs exposed to different transport times.

Twenty pigs were randomly divided into four groups of five pigs each (not transported - control, 1, 2 and 4h of transportation). A significant increase of ALT, AST and CK in the blood serum and acute parenchyma cell lesions were observed and those were characterized by acute degenerations in the heart and liver. Hsp27 expression levels increased significantly in the heart after 2h and in the liver after 4h of transportation, accompanying with the hsp27 mRNA increasing significantly in the heart and liver after 1h of transportation. αB-crystallin expression levels were fluctuant (not significantly) in the heart and liver during transporting, however, αB-crystallin mRNA increase notably in the heart after 1h and decrease significantly in the liver at 1 and 2h of transportation, respectively. In conclusion, the cellular damage to the heart and liver is highest after 1h of transportation, Hsp27 and αB-crystallin play dissimilar roles and show tissue-specific response in different tissues during transportation.

Anti-virus activity induced by polyinosinic:polycytidylic acid copolymer in in vitro chick embryo fibroblast cells.

The effect of a polyinosinic:polycytidylic acid copolymer (Poly I:C) on chick embryo fibroblast (CEF) cell viability, anti-Newcastle disease virus (NDV) activity and on IFN-alpha and IFN-beta levels were determined in vitro. Poly I:C inhibited NDV growth in CEF cells in the range of 30-500 microg/mL and over 0.75-12 h of treatment.The anti-NDV activity was maximal for 60 microg/mL of Poly I:C treatment at 12 h, after which it declined. The result of VSV protection assay which was used to determine IFN-activity showed that Poly I:C inhibited VSV growing on CEF cells, indicating that Poly I:C had immune-stimulating activities.The amount of IFN-beta in cell supernatant and the transcription levels of IFN-alpha mRNA and IFN-beta mRNA increased after Poly I:C treatment, indicating that the anti-viral activity related to the production of type I IFN. However Poly I:C also showed a toxic effect by inhibiting CEF cell proliferation in a dose-dependent and time-dependent manner.The results of this study indicate that the dosage level of Poly I:C must be carefully considered when used in a clinical setting.