Simple method for isolation of glyceraldehyde 3‐phosphate dehydrogenase and the improvement of myofibril gel properties

Porcine glycoliytic enzyme, glyceraldehyde 3‐phosphate dehydrogenase (G3PD) was prepared effectively by a combination of ethylene diamine tetra‐acetate (EDTA) pretreatment and affinity purification. After salting out of porcine sarcoplasmic proteins (SP) with ammonium sulfate at 75% saturation, the obtained supernatant (SP‐f3) was treated with EDTA, leaving G3PD in the supernatant (G3PD‐E) and most other SPs in the precipitate. At that time, the separation of G3PD‐E required more than 20 mmol/L EDTA. G3PD‐E was then subjected to affinity purification by batchwise method using blue‐sepharose CL‐6B, and purified G3PD (G3PD‐AP) was obtained using 2 mol/L potassium chloride (KCl) as an eluent. Texture analysis showed that the hardness, adhesiveness and gumminess of the myofibril gel at 0.2‐mol/L NaCl increased with the addition of G3PD‐AP. Scanning electron microscopy revealed that the G3PD‐AP reinforced the gel network of the myofibril. However, scanning electron micrograph analysis showed that the network‐structure of the gel by the addition of G3PD‐AP developed in a different manner from that by adding 0.6 mol/L NaCl. These results showed that glycolytic enzyme, G3PD, contributes to the improvement of the rheological properties of meat products.     

An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In Vitro

We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E₂ (PGE₂) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE₂ at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.     

Age and growth of Sebastes vulpes in the coastal waters of western Hokkaido, Japan

ABSTRACT:   Age and growth of Sebastes vulpes collected from the coastal waters of western Hokkaido were studied. Observation of the otolith margin verified that annuli (outer margins of the opaque zone) were produced chiefly from July to August. This period was associated with parturition and the birth season. The maximum age estimated by the surface method was 12 years but the oldest fish was aged at 35 years by the cross-section method. The surface method was inadequate for aging of S. vulpes older than 6 years because of the underestimation of age. No significant difference was found in the parameters of the growth equations between both sexes. The von Bertalanffy growth curve combined for both sexes was as follows: SL _t  = 358.6(1 − exp^{−0.156( t +0.820)}), where SL _t is standard length (mm) at age t (after parturition in years). It seems likely that S. vulpes grows slowly and lives longer than previously thought.     

Diel activity and vertical distribution of Lysianassoid amphipods dominant in Onagawa Bay, north-eastern Japan

ABSTRACT:   Baited traps were used over 24 h to reveal the diel activity and vertical distribution of three dominant lysianassoid amphipods ( Scopelocheirus onagawae , Anonyx omorii , Anonyx abei ) in Onagawa Bay, north-eastern Japan. The three species were the most dominant small crustacean scavengers in the bay. Most individuals were caught by traps deployed 0.3 m above the bottom, showing that they were active in the layers close to the sea bottom. No habitat segregation was found among species or developmental stages, suggesting that these scavengers rely on the food supply from bottom deposits. The three species increased remarkably in numbers of collected individuals at night, showing a rise in feeding activity. The two Anonyx species were collected only during the night, and S. onagawae was collected mostly during the night. The nocturnal activity appears to be an adaptation to reduce predation risks. However, juveniles of S. onagawae appeared even during the day, suggesting that feeding might take precedence over avoiding predation. Baited traps collected ovigerous females of S. onagawae but not of Anonyx species; the reason for the difference could not be determined.     

Granulosa cell tumor in 8 African pygmy hedgehogs (Atelerix albiventris)

A retrospective study involving eight African pygmy hedgehogs histopathologically diagnosed with granulosa cell tumors was conducted. The age at onset was 2.2–4.5 years, with a median age of 3.6 years. The most common clinical signs were hematuria and abdominal distension, which were observed in >50% cases. Exploratory laparotomy was performed in all cases, and ovariohysterectomy or excision of the abdominal mass was performed. Patients with only hematuria survived for >250 days after surgery, whereas those with initial ascites showed recurrence of ascites or tumor growth and survived for approximately 130 days after surgery. Intraperitoneal injection of carboplatin was performed in three recurrent cases. In one of these three cases, the tumor mass disappeared. Hence, carboplatin can be considered a potential antineoplastic drug for the treatment of granulosa cell tumors.