Comparative observation of freeze–thaw-induced damage in pig, rabbit, and human corneal stroma

Objective  Although variations exist between species with respect to outcomes after cryopreservation, little is known about the differences in the susceptibility of the corneal stroma to cryoinjury. We performed this study to investigate freeze–thaw-induced damage in keratocytes and collagen in rabbit, pig, and human corneas.
Animals studied  Rabbit, pig, and human.
Procedures  We prepared 250-μm-thick anterior stroma from rabbit, pig, and human corneas after scraping off the epithelium and endothelium. Each 250-μm-thick corneal stroma without epithelium was placed in a 50-mL tube, frozen with liquid N₂ for 15 min and taken out to thaw rapidly at 37 °C. This procedure of rapid freezing and thawing was repeated three times. Differences between the species with respect to cells and collagen structures were examined using hematoxylin–eosin (H&E) staining, terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay, and transmission electron microscopy (TEM). We orthotopically transplanted the pig and rabbit corneal transplants after the triple freeze–thaw cycle into rabbit eyes and evaluated graft survival.
Results  On gross examination, rabbit corneas became opaque after the triple freeze–thaw procedure, while pig and human corneas remained transparent. Histologically, keratocytes were apoptotic on TUNEL assay and TEM in rabbit, pig, and human corneas. Collagen fibrils were fragmented and the arrangement of collagen fibrils was severely disturbed in rabbit corneas on H&E staining and TEM; collagen was well preserved in pig and human corneas. Rabbit corneal stroma underwent autolysis after transplantation, whereas the pig corneal stroma remained clear for 1 month.
Conclusions  Our study showed that rabbit corneal stroma was more susceptible to freeze–thaw injury than pig and human corneas.     

Long‐term changes in recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) and environmental conditions around Japan

Recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) from 1952 to 2014 was examined by a sequential regime shift detection method. The regime shifts in recruitment were detected in 1957, 1972, 1980, 1994 and 2009. The durations of regime shift ranged from 8–15 years and averaged 13.0 years. In both the total (1952–2014) and data rich (1980–2014) periods, negative relationships were found between recruitment and the Pacific Decadal Oscillation in autumn, and positive relationships were found between recruitment and sea surface temperature (SST) anomalies in the northern part of the East China Sea, in the southwestern part of the Sea of Japan, and in the waters off Shikoku and Tokai in summer and autumn. The 1994 and 2009 regime shifts in recruitment occurred in the same years as shifts in SST anomalies in the northern part of the East China Sea in summer. These results suggest that the ocean conditions in the northern part of the East China Sea are closely related to recruitment of Pacific bluefin tuna, and that the warmer conditions result in higher recruitment of the species.     

Hypereosinophilic syndrome in two cats

Two cats showing chronic vomiting, diarrhea and weight loss were found to have leukocytosis with marked eosinophilia. Both cats were diagnosed with hypereosinophilic syndrome by the findings of increased eosinophils and their precursors in the bone marrow, eosinophilic infiltration into multiple organs, and exclusion of other causes for eosinophilia. Although cytoreductive chemotherapy with hydroxycarbamide and prednisolone was performed, these two cats died 48 days and 91 days after the initial presentation.     

A comparison of two combinations of xylazine-ketamine administered intramuscularly to alpacas and of reversal with tolazoline

ObjectiveTo evaluate the anesthetic and cardiorespiratory effects of two doses of intramuscular (IM) xylazine/ketamine in alpacas, and to determine if tolazoline would reduce the anesthetic recovery time.Study designProspective randomized crossover study.AnimalsSix castrated male alpacas.MethodsEach alpaca received a low dose (LD) (0.8 mg kg⁻¹ xylazine and 8 mg kg⁻¹ ketamine IM) and high dose (HD) (1.2 mg kg⁻¹ xylazine and 12 mg kg⁻¹ ketamine IM) with a minimum of one week between trials. Time to sedation, duration of lateral recumbency and analgesia, pulse rate, respiratory rate, hemoglobin oxygen saturation, arterial blood pressure, blood-gases, and the electrocardiogram were monitored and recorded during anesthesia. With each treatment three alpacas were randomly selected to receive tolazoline (2 mg kg⁻¹ IM) after 30 minutes of lateral recumbency.ResultsOnset of sedation, lateral recumbency and analgesia was rapid with both treatments. The HD was able to provide ≥30 minutes of anesthesia in five of six alpacas. The LD provided ≥30 minutes of anesthesia in three of six alpacas. Respiratory depression and hypoxemia occurred with the HD treatment during the first 10 minutes of lateral recumbency: two animals were severely hypoxemic and received nasal oxygen for 5 minutes. Heart rate decreased, but there were no significant changes in arterial blood pressure. Tolazoline significantly shortened the duration of recumbency with the HD.ConclusionsThe HD provided more consistent clinical effects in alpacas than the LD. Intramuscular tolazoline shortened the duration of lateral recumbency in alpacas anesthetized with the HD combination.Clinical relevanceBoth doses of the combination were effective in providing restraint in alpacas and the duration of restraint was dose dependent. Supplemental oxygen should be available if using the HD and IM administration of tolazoline will shorten the recovery time.     

Effect of Addition of Different Acetic Acid Concentrations on the Quality of Marinated Herring

This study investigated the effect of acetic acid concentrations from 3 to 8% on the physicochemical and sensory quality of marinated Atlantic and Baltic herring from fresh and frozen-thawed material. The increased concentration of acetic acid was found to exert a strong impact on decreased sensory scores, weight yield, and pH value as well as on an increased brightness parameter of meat. In turn, moisture, protein hydrolysis products (PHB), total volatile bases (TVB) content, and hardness parameter were affected by the higher addition of acid to a lesser extent. Enhanced protein hydrolysis was observed only at acetic acid concentration of 5% or 5–8%, whereas there was a decrease in the content of TVB only at an acid concentration of up to 4–5%. The marinated frozen-thawed fillets, compared to the fresh ones, required a greater addition of acid in the brine, by 1.5%, to reach a similar pH value. The freezing of fish contributed also to a decrease of PHB and free hydroxyproline, along with an increased concentration of acetic acid. The marinades from frozen-thawed herring contained more TVB, had worse color, and hardness parameters, as well as less yield.     

Schisandrin B protects against solar irradiation-induced oxidative injury in BJ human fibroblasts

The effects of schisandrin B (Sch B) and its analogs on solar irradiation-induced oxidative injury were examined in BJ human fibroblasts. Sch B and schisandrin C (Sch C) increased cellular reduced glutathione (GSH) level and protected against solar irradiation-induced oxidative injury. The photoprotection was paralleled by decreases in the elastases-type protease activity and matrix-metalloproteinases-1 expression in solar-irradiated fibroblasts. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production. The results suggest that by virtue of its pro-oxidant action and the subsequent glutathione antioxidant response, Sch B or Sch C may offer the prospect of preventing skin photo-aging.     

Monoclonal antibody-based competitive ELISA for simultaneous detection of rinderpest virus and peste des petits ruminants virus antibodies

An experimental competitive enzyme-linked immunosorbent assay (morbillivirus cELISA) using a recombinant N antigen (rRPV N) expressed in a baculovirus and a ruminant morbillivirus (RPV and PPRV)-specific monoclonal antibody (P-13A9) was developed for simultaneous detection of rinderpest virus (RPV) and peste des petits ruminants virus (PPRV) antibodies and its diagnostic performance was evaluated. A set of known reference antisera against RPV and PPRV belonging to different lineages, experimental sera from cattle vaccinated for a RPV of Asian lineage, and field sera from cattle and sheep/goat populations known to be positive (West Africa) and negative (Korea) for RPV and PPRV were used for the evaluation. Morbillivirus cELISA results on the panel of experimental RPV and PPRV antisera showed high correlation (r=0.97) between the whole virus and the rRPV N antigens, suggesting that the rRPV N contains a ruminant morbillivirus-specific antigenic determinant recognized by the P-13A9 and it may be suitable as an ELISA antigen in place of the whole virus. Morbillivirus cELISA detected anti-RPV and anti-PPRV antibodies in all reference RPV and PPRV antisera containing VN titers >/=1:8, suggesting that the assay can simultaneously detect antibodies against RPV and PPRV. Anti-RPV antibody was detected by morbillivirus cELISA in vaccinated cattle as early as the VNT and continued to be detectable by both the cELISA and the VNT until termination of the study. When applied to field samples from Africa, morbillivirus cELISA showed good agreement with a RP cELISA kit (kappa value of 0.86) in bovine sera and with a peste des petits ruminant cELISA kit (kappa value of 0.81) in caprine/ovine sera. Usefulness of morbillivirus cELISA using the rRPV N protein was discussed.     

Cross-protective immunity and the serological classification system for Bacteroides nodosus

The relationship between the serological classification system for serogroup B and for serogroup H of Bacteroides nodosus and cross-protection between subgroups within these serogroups was examined. Protection against ovine footrot following vaccination was achieved against other subgroup strains provided sufficient cross-reactive antibody was induced by shared pilus antigens. Within serogroup B, better cross-protection against one subgroup was obtained with a pili vaccine than a whole cell vaccine which correlated with higher pilus antibody titres induced by the former. For serogroup H, a lack of cross-protection and serological reactivity between subgroups was demonstrated, which indicates that the prototype strain of subgroup H2 should be designated a new serogroup.     

Effects of administration of IH901, a ginsenoside intestinal metabolite,on muscular and pulmonary antioxidant functions after eccentric exercise

This study was conducted to investigate whether administration of IH901, a ginseng intestinal metabolite, ameliorates exercise-induced oxidative stress while preserving antioxidant defense capability in rat skeletal muscles and lung. Eight adult male Sprague-Dawley rats per group were randomly assigned to the resting control, exercise control, resting with IH901 (25, 50, and 100 mg/kg) consumption (R/IH901), or exercise with IH901 (25, 50, and 100 mg/kg) consumption (E/IH901) group. The trained groups ran 35 min 2 days/week for 8 weeks. To analyze the IH901-training interaction, serum biochemical analysis, lipid peroxidation, citrate synthase, protein oxidation, antioxidant and superoxide dismutase in skeletal muscles and lung tissue were measured. Compared to the exercise control group, animals that consumed IH901 had significantly increased exercise endurance times (p < 0.05) and decreased plasma creatine kinase and lactate dehydrogenase levels (p < 0.05), while those in the E/IH901 groups had increased citrate synthase and anti-oxidant enzymes and decreased lipid peroxidation and protein oxidation (p < 0.05). In conclusion, IH901 consumption in aging rats after eccentric exercise has beneficial effects on anti-inflammatory and anti-oxidant activities through down-regulation of pro-inflammatory mediators, lipid peroxidation, and protein oxidation and up-regulation of anti-oxidant enzymes.