Histological and immunohistochemical evaluation of granulosa cells during different stages of folliculogenesis in bovine ovaries.

Bovine granulosa cells (GC) vary in their morphological aspect during different stages of folliculogenesis. In this study, 10 morphologically normal bovine ovaries were collected to study the structural aspects of different stages of GC using intermediate filament protein antibodies including cytokeratin AE1/AE3 (AE1/AE3), vimentin, nectin-4 and desmin. Hormonal immunolocalization was assessed using the immunomarkers anti-Müllerian hormone (AMH) and inhibin alpha. In addition, tumour markers and proliferation markers using c-erbB-2 oncoprotein and proliferating cell nuclear antigen, respectively, were investigated. The immunolabelling of AE1/AE3 in GC was strongest in the early follicle stage and gradually decreased when reaching the Graafian follicle stage. Its immunolabelling increased again as the stage progressed from stage I to stage III. The immunolabelling of inhibin alpha was inversely proportional to that of AE1/AE3 in the developing ovarian follicles as their immunolabelling is opposite to each other during folliculogenesis. AMH was immunopositive in almost all GC stages in different intensities and percentages, except for some negative staining in the atretic IV follicles. The atretic IV follicle is a unique type of atretic follicle that shows Call-Exner body formation, which was mainly found in older cows in this study. The distinct patterns of immunoreactivity for various types of immunomarkers in the different GC stages will play an important role in diagnostic assistance of various follicle conditions, including cystic ovaries and GC tumours.

Disseminated Pneumocystosis in a Toy Poodle.

A 1-year and 7-months-old neutered male toy poodle was presented with persistent respiratory distress, gradual weight loss and melaena. Thoracic radiography showed an unstructured interstitial lung pattern. Histopathological examination of tissues collected at necropsy examination revealed disseminated infection by Pneumocystis carinii. The organisms were detected in the lungs, lymph nodes, liver, heart, kidneys, spleen, gastrointestinal tract and pancreas. In the lungs, the organisms were present in the alveolar space and interstitial tissue, and calcified foci containing P. carinii were observed. The presence of the organism in non-thoracic lymph nodes provided evidence of lymphogenous spread. A definitive diagnosis of disseminated pneumocystosis was achieved through the use of Grocott methenamine silver staining, immunohistochemistry (IHC) and polymerase chain reaction for P. carinii. Depletion of cells expressing immunoglobulin (Ig)A and IgG was confirmed by IHC of lymphoid tissue, suggesting possible underlying immunodeficiency.

Appearance of US-like porcine epidemic diarrhoea virus (PEDV) strains before US outbreaks and genetic heterogeneity of PEDVs collected in Northern Vietnam during 2012-2015.

Porcine epidemic diarrhoea virus (PEDV) is the aetiologic agent of porcine epidemic diarrhoea (PED), a highly contagious enteric disease that is threatening the swine industry globally. Since PED was first reported in Southern Vietnam in 2009, the disease has spread throughout the country and caused substantial economic losses. To identify PEDVs responsible for the recent outbreaks, the full-length spike (S) gene of 25 field PEDV strains collected from seven northern provinces of Vietnam was sequenced and analysed. The sequence analysis revealed that the S genes of Vietnamese PEDVs were heterogeneous and classified into four genotypes, namely North America and Asian non-S INDEL, Asian non-S INDEL, new S INDEL and classical S INDEL. This study reported the pre-existence of US-like PEDV strains in Vietnam. Thirteen Vietnamese variants had a truncated S protein that was 261 amino acids shorter than the normal protein. We also detected one novel variant with an 8-amino acid insertion located in the receptor-binding region for porcine aminopeptidase N. Compared to the commercial vaccine strains, the emerging Vietnamese strains were genetically distant and had various amino acid differences in epitope regions and N-glycosylation sites in the S protein. The development of novel vaccines based on the emerging Vietnamese strains may be contributive to the control of the current PED outbreaks.

Non-tuberculous Mycobacteriosis with T-cell Lymphoma in a Red Panda (Ailurus fulgens).

A 9-year-old male red panda (Ailurus fulgens) became emaciated and died. Necropsy examination revealed systemic lymphadenomegaly. The liver, lungs and left kidney contained multifocal yellow nodules. Microscopical examination revealed granulomatous inflammation in the liver, lungs, kidney, spleen and lymph nodes, with numerous acid-fast bacilli. Sequencing of genetic material isolated from the tissues classified the pathogen as Mycobacterium gastri. Lymphoma was found in the liver, lungs, kidney and lymph nodes. The neoplastic cells were strongly labelled for expression of CD3, Ki67 and proliferating cell nuclear antigen by immunohistochemistry. This is the first report of M. gastri infection with T-cell lymphoma in a red panda.

Burn injury to trunk of rat causes denervation-like responses in the gastrocnemius muscle.

Thermal injury results in dystropic changes in skeletal muscle and abnormal pharmacological responses to neuromuscular relaxants, each of which suggests a denervation-like phenomenon. In the rat thermal injury model we examined whether, as in denervation states, increases in nicotinic acetylcholine receptors (AChR) and hyposensitivity to d-tubocurarine (dTc) are found. While anesthetized, thermal injury was imposed to trunk only. At 10, 14, and 21 days after injury the effective doses of dTc for left gastrocnemius tension suppression to 95% of control tension (ED95) were 0.213 +/- 0.039, 0.305 +/- 0.070, and 0.214 +/- 0.032 mg/kg, respectively. These values were significantly higher (P less than 0.05) than control values (0.155 +/- 0.006 mg/kg). The AChR concentrations in the left gastrocnemius, quantitated by 125I-alpha-bungarotoxin binding, increased at 10, 14, and 21 days to 182 +/- 20% (P less than 0.001), 166 +/- 22% (P less than 0.03), and 164 +/- 18% (P less than 0.001) of control, respectively. AChR concentrations in the right gastrocnemius also increased subsequent to thermal injury. Changes in effective dose of dTc for 50 and 95% twitch suppression in the left gastrocnemius correlated significantly with changes in AChR concentrations for the same muscle (r = 0.73 and 0.81, P less than 0.001, respectively). This study confirms the hypothesis that the systemic effects of thermal injury include an increase in AChR at sites distant from thermal injury, which may account for the skeletal muscle dysfunction and aberrant responses to neuromuscular relaxants.

Concentration-dependent interaction of theophylline with d-tubocurarine.

The interaction of theophylline with d-tubocurarine chloride (dTC) was examined in rabbits. After steady-state subtherapeutic (less than 10 mg/l), therapeutic (10-20 mg/l), and toxic (greater than 20 mg/l) concentrations of theophylline, dose-response curves for dTC were determined and compared with controls that received no theophylline. At therapeutic concentrations of theophylline the effective dose for 50% inhibition of twitch (ED50) for dTC (mean +/- SE, 0.115 +/- 0.016 mg/kg) was significantly shifted to the left in comparison with the control (0.165 +/-0.008 mg/kg). The ED50 of dTC for the subtherapeutic group was 0.143 +/- 0.011 mg/kg, which was less than the control but not of statistical significance (P = 0.1). The ED50 for the toxic theophylline group was 0.168 +/- 0.003 mg/kg, which was not significantly different from controls but significantly different from the theophylline therapeutic and subtherapeutic groups. Thus, toxic concentrations of theophylline reversed the potentiating effects of therapeutic and subtherapeutic concentrations of dTC dose-response curves. Therefore, depending on concentration, theophylline exhibits a biphasic interaction with dTC. Surgical patients on theophylline may require less dTC intraoperatively. More importantly, the use of theophylline in the postoperative period to reverse anesthetic effects may result in recurarization.

PaO2 and intratracheal pressure in oscillatory ventilation in experimental respiratory failure.

In order to study the effect of high frequency ventilation on the pulmonary gas exchange in respiratory failure, we measured the PaO2 and other pulmonary gas exchange parameters for ventilatory rates of 0.5, 1.2, 4.8, and 16 Hz in dogs, to which acute respiratory failure was created by intravenous infusion of oleic acid. Either the mean intratracheal pressure or the end-tidal intratracheal pressure was controlled. A loud-speaker ventilator of our own design was modified so as to enable variation of the intratracheal pressure. Ventilation was measured using an ultrasonic instrument which counts the number of turbulent eddies. The tidal volume was set slightly higher than that was determined for healthy animals, but the resulting PaCO2 values were consistently higher than normal when PaO2 values were low. With the mean intratracheal pressure kept constant at 5, 10, and 15 cm H2O, PaO2 values with the FIO2 of 1 were between 41 and 46 at mPit of 5, 65 and 82 at 10 cm H2O, 278 and 423 at 15 cm H2O. No increase in PaO2 was observed with the increase in respiratory frequencies. If anything, a slight reduction in PaO2 at 8 and 16 Hz was observed, though not statistically significant. With the end-tidal intratracheal pressure constant, PaO2 varied but again correlated well with the mPit.

[The effect of high frequency jet ventilation on intracranial pressure in the patients with severe head injury].

The effect of high frequency jet ventilation (HFJV) on intracranial pressure (ICP) in the series of seven patients with severe head injury was studied. These patients received mechanical ventilation for respiratory failure accompanied with neural damage or circulatory shock and for ICP control by decreasing PaCO2. ICP was measured continuously by subarachnoid catheter method connected to Gould P-50 strain-gage transducer and controlled by hyperventilation and/or osmotic diuretics during acute phase. Arterial pressure, central venous pressure and intratracheal pressure were measured continuously in the same way. Arterial blood gases were analysed by BG-I of Technicon Co. Ltd. At the weaning from respirator, two methods were applied. One was conventional IMV (with PEEP) and/or CPAP by using IMV mode of Servo 900 respirator or T-piece together with PEEP valve and another was new-fashioned HFJV. Mean intratracheal pressure (mPit) was fixed at 0 mmHg (control value), 5, 10 and 15mmHg in both methods by adjusting PEEP valve or driving pressure of HFJV. The result is that in the same patients ICP was significantly lower during HFJV than IMV/CPAP when mPit was 5 and 10mmHg. Only three cases having been studied at 15mmHg of mPit because circulatory condition would not permit it, significant difference of ICP between HFJV and IMV/CPAP was not detected. PaCO2, being suspected to be the cause of lower ICP of HFJV group, was studied too. But blood gas analysis showed no significant difference neither PaCO2 nor PaO2. The fluctuation of CVP reflecting the change of intrathracic pressure was smaller in HFJV group than in IMV/CPAP group.(ABSTRACT TRUNCATED AT 250 WORDS)

[A classification of shock based on hemodynamic profiles].

Shock is a syndrome of numerous etiologies. The treatment of shock syndrome depends on the etiology and circulatory pathophysiology. From the viewpoint of circulatory physiology, the mechanism of circulatory failure can be attributed to three factors: blood volume; cardiac contractility; and vascular resistance. Balloon-tipped pulmonary artery (Swan-Ganz) catheters can distinguish between these factors in the clinical setting and allows physicians to treat shock patients most appropriately and logically. The author proposes a treatment-oriented classification of shock based on Swan-Ganz catheter findings and divides shock into three types: hypovolemic; cardiogenic; and low-resistant shock. All clinicians who treat shock must have qualified knowledge of and experience in handling this beneficial but invasive monitoring device.

Membranoproliferative glomerulonephritis in a calf with nephrotic syndrome.

A 2-month-old Japanese black calf was presented with a history of weight loss, exophthalmos and subcutaneous oedema of the brisket. Urinalysis and serum biochemistry showed proteinuria and hypoproteinaemia suggestive of nephrotic syndrome. Microscopically, lesions in the kidney were characterized by proliferation of mesangial cells and diffuse thickening of the glomerular basement membranes with the appearance of double contours. Immune complex deposits were confirmed by electron microscopy and immunofluorescence using reagents specific for bovine immunoglobulin G, complement factor C3 and bovine viral diarrhoea virus (BVDV). Consequently, the glomerular lesion in this case was diagnosed as membranoproliferative glomerulonephritis. BVDV type 1 was detected in serum by nested reverse transcriptase polymerase chain reaction. Viral antigen was also identified in the glomeruli by immunofluorescence. These results suggest that BVDV may have been the cause of immune complex glomerulonephritis in this calf.

Oral administration of cilnidipine to patients with hypertensive intracerebral hemorrhage in the acute stage: significance and role of an N-type calcium channel blocker.

BACKGROUND: Elevated blood pressure (BP) causes rebleeding or enlargement of intracerebral hematomas. AIMS: How a long-acting oral calcium channel blocker, cilnidipine, could control BP in the acute stage of cerebral hemorrhage was evaluated. METHODS AND RESULTS: Cilnidipine given within 3 days of hospitalization has more benefit than cilnidipine given after 4 days of hospitalization; it can reduce the amount of intravenous nicardipine, and it can help to maintain the BP below 80% of the initial BP. Surgical removal of the hematoma has no benefit in reducing the amount of intravenous nicardipine and maintaining the BP below 80% of the initial BP. CONCLUSION: In order to reduce the total amount of intravenous nicardipine and to maintain the BP below 80% of the initial BP, oral administration of a long-acting N-type calcium channel blocker, cilnidipine, is useful and important, independent of whether the hematomas are surgically removed.

Burn injury to rat increases nicotinic acetylcholine receptors in the diaphragm.

Thermal injury induces aberrant responses to neuromuscular (NM) blocking drugs, and it has been speculated that an increase in nicotinic acetylcholine receptors (AchR) may contribute to the altered response. Using the diaphragmatic muscle as a representative of skeletal muscle, the changes in AchR were examined. The diaphragm, rather than limb muscles, was chosen to avoid the effects of wound contracture-induced immobilization and denervation of limb, which can also increase AchR in skeletal muscle. Study of changes in diaphragm also tested the hypothesis that increase in AchR are the result of a generalized systemic effect, and not limited to area of burn. Following a 45-55% body surface area thermal injury to the trunk (not limbs) of rats, AchR changes in the diaphragm were studied at 10, 14, 21, and 28 days after injury and compared to uninjured (control) rats. The AchR changes in the diaphragmatic muscle was assayed using 125I-alpha-bungarotoxin as the specific ligand. At 10, 14, and 21 days after thermal injury, the animals had an arrest in weight growth and the AchR concentration was increased to (mean +/- SE) 155 +/- 15% (P less than 0.02), 160 +/- 16% (P less than 0.003), and 141 +/- 16% (0.05 less than P less than 0.1), respectively, compared to control. At 28 days, probably because of wound healing and burn wound contracture, the size of thermally injured area was significantly (P less than 0.001) reduced to 19 +/- 4% of body surface area, and weight increased (P less than 0.001) compared to preburn weight.(ABSTRACT TRUNCATED AT 250 WORDS)