Morphological analysis of cell distribution and network structure via gap junctions in swine corneal stroma

Because of corneal transplantation limitations, there is a need for cornea-specific regenerative medicine. The development of such regenerative medicine has been delayed because of the complex and unique structure of the corneal stroma. Few studies have explored the corneal stroma cell distribution and cell types in vivo. This study investigated regional differences in morphological characteristics and distributions of corneal keratocytes and immunocompetent cells in the corneal stroma to clarify their functions and structural characteristics. The porcine eyeballs were subjected to light microscopy, transmission electron microscopy, scanning electron microscopy, and immunofluorescence staining analyses. Corneal cells were primarily located in the limbus, rather than the center of the cornea; the long keratocyte diameter was largest on the epithelial side of the corneal limbus, while the short diameter was largest on the endothelial side of the central cornea. Moreover, there were significantly more corneal cells on the epithelial side than on the endothelial side in both the central and limbus areas. Gap junctions between cells in the corneal stroma were present on the surfaces of cytoplasmic processes. Many cytoplasmic processes were scattered throughout the corneal stroma; they were connected both vertically and horizontally, forming an intercellular network. Additionally, immunocompetent cells on the epithelial side suggested to participate in this network via gap junctions. The morphology of keratocytes and immunocompetent cells on the epithelial side suggests that they play important roles in corneal homeostasis.     

Effects of Nitrogen Fertilization on Stream Chemistry of Japanese Forested Watersheds

Steam chemistry was investigated from May 1991 through April 1992 for 13 Japanese forested watersheds and from May 1990 through August 1994 for two of these watersheds. Nine watersheds were treated over different periods (1983–1991) with different amounts of N (nitrogen) fertilizer as urea and ammonium salts. Total N additions ranged from 20 to 375 kg ha^-1. There were no distinct seasonal differences in stream NO₃ ^- concentrations in either the treated or untreated watersheds, but concentrations tended to be somewhat higher during periods of high discharge. The annual average NO₃ ^- concentrations in streams had a significant, positive (p < 0.001, r = 0.84) relationship to the total amount of N applied from 1985–1991. The application of 330 kg N ha^-1 raised annual average stream NO₃ ^- concentration to about 300 μeq L^-1 compared to less than 160 μeq L^-1 in untreated watersheds. The concentrations of K⁺, Ca²⁺, and Mg2+ in stream water also increased in those watersheds with high rates of N fertilizer as a result of nitrification that increased the generation of the mobile nitrate anion. The lack of seasonality in stream NO₃ ^- concentrations and the large rates of N loss with N addition both suggest that these watersheds were ‘nitrogen saturated’     

Determinants of the daily rhythm of blood fluidity

Background  

Numerous processes in the living body exhibit daily rhythmicity. In this study, we characterized a daily rhythm of blood fluidity and identified its determinants.     

Cost-effectiveness of diagnostic strategies using quantitative real-time PCR and bacterial culture to identify contagious mastitis cases in large dairy herds

Diagnostic strategies to detect contagious mastitis caused by Mycoplasma bovis, Staphylococcus aureus, and Streptococcus agalactiae in dairy herds during an outbreak have been minimally studied with regard to cost and diagnostic sensitivity. The objective of this cross-sectional study was to compare the cost-effectiveness of diagnostic strategies for identification of infected cows in two California dairy herds during contagious mastitis outbreaks.     

Assessment of Alloxan-Induced Diabetic Rats as a Periodontal Disease Model Using a Selective Cyclooxygenase (COX)-2 Inhibitor

Several recent studies have reported that alloxan-treated rats with long-term hyperglycemia can develop naturally occurring periodontal disease (PD). Our previous studies detected dental caries in the same model. Therefore, these two lesions of different etiologies are expected to occur concurrently. In this study, we evaluated the use of diabetic rats as a PD model by employing a selective COX-2 inhibitor reported to be effective against PD. Six-week-old female F344 rats were divided into 3 groups: intact rats (control), alloxan-induced diabetic rats fed a standard diet (AL) and alloxan-induced diabetic rats fed a diet containing 0.01% etodolac (AL+Et). The animals were euthanized at 26 weeks of age, and their oral tissues were examined histopathologically. Gingivitis, marginal periodontitis and alveolar bone resorption were markedly enhanced along with dental caries in the AL group compared with the control group. However, the COX-2 inhibitor had no effect on periodontal inflammation in the AL+Et group. In addition, in the AL group, periodontitis was notably nonexistent around the normal molars, and gingivitis was scarcely worse than that in the control group. In the diabetic rats, the progression of periodontal inflammation was closely correlated with the severity of adjacent dental caries, and marginal periodontitis was frequently continuous with apical periodontitis. In conclusion, an alloxan-induced diabetic rat is not a model of PD but of dental caries. It is probable that in this model, hyperglycemia may enable crown caries to progress to apical periodontitis, while the associated inflammation may rostrally expand to surrounding periodontal tissue.     

The effect of body position on the 'hanging drop' method for identifying the extradural space in anaesthetized dogs

ObjectivesTo assess the accuracy of the ‘hanging drop method’ for identifying the extradural space in anaesthetized dogs positioned in sternal or lateral recumbency.Study designProspective randomized-experimental study.AnimalsSeventeen clinically healthy adult dogs, 10 females and seven males weighing 8.4–26.2 kg.MethodsDogs were positioned in either sternal (n = 8) or lateral (n = 9) recumbency under general anaesthesia. A 20 SWG spinal needle pre-filled with 0.9% saline was advanced through the skin into the lumbosacral extradural space and the response of the saline drop recorded, i.e. whether it: 1) was aspirated from the hub into the needle; 2) remained within the hub, or 3) moved synchronously with i) spontaneous respiration, ii) heart beat or iii) manual lung inflation. The position of the needle tip was ultimately determined by positive contrast radiography.ResultsOne dog positioned in lateral recumbency was excluded from the study because bleeding occurred from the needle hub. Saline was aspirated into the needle in seven of eight dogs held in sternal recumbency but in none of the dogs positioned in lateral recumbency. Accurate needle tip placement in the extradural space was confirmed by positive contrast radiography in all dogs.Conclusion and clinical relevanceThe ‘hanging drop’ method, when performed with a spinal needle, appears to be a useful technique for identifying the location of the extradural space in anaesthetized medium-sized dogs positioned in sternal, but not in lateral recumbency. The technique may yield ‘false negative’ results when performed in dogs positioned in sternal recumbency.     

Laparoscopic cryptorchidectomy in standing bulls

Laparoscopic cryptorchidectomy without insufflation was applied in 10 standing bulls aged 3 to 15 months. Nine bulls were preoperatively pointed out intra-abdominal testes by computed tomography. Preoperative fasting for a minimum of 24 hr provided laparoscopic visualization of intra-abdominal area from the kidney to the inguinal region. Surgical procedure was interrupted by intra-abdominal fat and testis size. It took 0.6 to 1.5 hr in 4 animals weighing 98 to 139 kg, 0.8 to 2.8 hr in 4 animals weighing 170 to 187 kg, and 3 and 4 hr in 2 animals weighing 244 and 300 kg to complete the cryptorchidectomy. In conclusion, standing gasless laparoscopic cryptorchidectomy seems to be most suitable for bulls weighing from 100 to 180 kg.     

The unique physiological features of the broiler pectoralis major muscle as suggested by the three-dimensional ultrastructural study of mitochondria in type IIb muscle fibers

Typical skeletal muscles are composed of mixed muscle fiber types, which are classified as slow-twitch (type I) and fast-twitch (type II) fibers, whereas pectoralis major muscles (PMs) in broiler chickens are 100% composed of type IIb fast-twitch fibers. Since metabolic properties differ among muscle fiber types, the combination of muscle fiber types is involved in physiological functions and pathological conditions in skeletal muscles. In this study, using serial block-face scanning electron microscopy, we compared three-dimensional (3D) mitochondrial properties in type IIb fibers in broiler PMs and those in type I fibers of broiler gastrocnemius muscles (GMs) heterogeneously composed of slow- and fast-twitch muscle fibers. In type I fibers in the GMs, elongated mitochondria with numerous interconnections to form a substantial network among myofibrils were observed. Along with lipid droplets sandwiched by mitochondria, these features are an adaptation to effective oxidative respiration and constant oxidative damage in slow-twitch muscle fibers. In contrast, type IIb fibers in the PMs showed small and ellipsoid-shaped mitochondria with few interconnections and no lipid droplets, forming a sparse network. The mitochondrial spatial network comprises of active mitochondrial dynamics to reduce mitochondrial damage; therefore, type IIb fibers possess physiologically low capacity to maintain mitochondrial wellness due to static mitochondrial dynamics. Based on 3D mitochondrial properties, we discussed the contrasting physiological functions between type I and IIb fibers and proposed a high contractile power and low stress resistance as unique physiological properties of broiler PMs.     

Nutrient Content in Buckwheat Milling Fractions

Buckwheat seeds (Fagopyrum esculentum Moench) were milled into 23 fractions: seven fine flours, three coarse flours, four small semolina, two big semolina, six bran, and one husk fraction. A considerable variation in gross chemical composition was found among the milling fractions. The protein content varied from 4.4 to 11.9% (db) in flours and from 19.2 to 31.3% in bran fractions; starch varied from 91.7 to 70.4% in flours and from 42.6 to 20.3 in bran. The percentage of soluble dietary fiber contained in total dietary fiber was higher in flours than in semolina and bran fractions. Ash, Fe, P, tannin, phytate content, and color were also investigated. A unique distribution of phytate was found in starch. Correlation is significantly positive in husk, bran, and semolina fractions, while correlation is significantly negative in flour fractions. Depending on technological or nutritional demands, appropriate fractions may be chosen to achieve the desired end‐use product.     

Arterial blood gas anomaly in canine hepatobiliary disease

Arterial blood gas analysis is an important diagnostic and monitoring tool for respiratory abnormalities. In human medicine, lung complications often occur as a result of liver disease. Although pulmonary complications of liver disease have not been reported in dogs, we have frequently encountered hypoxemia in dogs with liver disorders, especially extrahepatic biliary obstruction. In addition, respiratory disorders account for 20% of perioperative fatalities in dogs. Therefore, in this study, we evaluated the respiratory status in dogs with hepatobiliary disease by arterial blood gas analysis. PaO₂ and PaCO₂ were measured. Alveolar-arterial oxygen difference (AaDO₂), the indicator of gas exchange efficiency, was calculated. Compared to healthy dogs (control group), hepatobiliary disease dogs had significantly lower PaO₂ and higher AaDO₂. Hypoxemia (PaO₂ of ≤80 mmHg) was observed in 28/71 dogs with hepatobiliary disease. AaDO₂ was higher (≥30 mmHg) than the control group range (11.6 to 26.4 mmHg) in 32/71 hepatobiliary disease dogs. By classifying type of hepatobiliary disease, dogs with extrahepatic biliary obstruction and chronic hepatitis showed significantly lower PaO₂ and higher AaDO₂ than in a control group. Dogs with chronic hepatitis also had significantly lower PaCO₂. The present study shows that dogs with hepatobiliary disease have respiratory abnormalities more than healthy dogs. Preanesthetic or routine arterial blood gas analysis is likely beneficial to detect the respiratory abnormalities in dogs with hepatobiliary disease, especially extrahepatic biliary obstruction and chronic hepatitis.