Platelet activating factor enhances the acrosome reaction, fertilization in vitro by subzonal sperm injection and resulting embryonic development in the rabbit

This study was conducted to investigate the effect of plateletactivating factor (PAF) on the acrosome reaction and fertilizingcapacity of spermatozoa, and development of the resulting embryosin the rabbit. Rabbit spermatozoa were exposed to PAF, Iyso-PAF,or high ionic strength medium (HIS) prior to subzonal sperminjection (SUZI) into 326 mature oocytes, or morphological assessmentof the acrosome reaction. The rates of fertilization and blastocystformation were compared among the three treatment groups. Acrosomereaction was assessed by fluorescein isothiocyanate-conjugatedPisum sativum agglutinin (FITC–PSA) staining and electronmicroscopy. PAF-treated spermatozoa fertilized the oocytes ata significantly higher rate (56.1%) than did lyso-PAF-(36.8%,P< 0.01) or HIS- (38.2%, P < 0.05) treated spermatozoa.The embryos produced by PAF-treated spermatozoa showed significantlyhigher blastocyst formation rates (34.0%) than lyso-PAF- (8.6%,P < 0.050) or HIS-(8.8%, P< 0.05) treated spermatozoa.FITC-PSA staining demonstrated a significantly higher incidenceof acrosome reaction in PAF-treated spermatozoa (45.8%) thanin Iyso-PAF- (28.0%, P < 0.01) or HIS- (34.9%, P < 0.01)treated spermatozoa. Acrosome reaction of PAF-treated spermatozoawas also confirmed by electron microscopy. PAF treatment ofspermatozoa enhances fertilizing capacity for SUZI possiblyby augmenting the acrosome reaction. Enhanced embryonic developmentwas also found in the oocytes fertilized by SUZI of PAF-treatedspermatozoa.     

Basal cell adenoma with parallel tubules in stromal cells of the parotid gland

A case of basal cell adenoma in the right parotid region of a 51 years old male was reported. The tumor measured 2.5 cm x 3 cm, was spherical and covered with a fibrous capsule. Histologically, it was a tubular monomorphic adenoma with scant edematous interstitial tissue. The stromal cells stained positively by the PAP method using anti-S-100 protein serum. Electron microscopically, the tumor cells forming tubular had many microvilli at the luminal surface, many filaments in the cytoplasm and well developed desmosomes in the intercellular junctions. Ordinary intracellular organelles of the tumor cells were small in number, and their nuclei were oval with shallow indentation. In the dilated rough endoplasmic reticulum of the stromal cells, many straight parallel tubules were found. The tubules measured from 15 nm to 25 nm thick and 3.5 micrometers long in the longitudinal sections and from 25 nm to 30 nm in diameter with electron lucent core and poor coat in the cross sections. Other cell organelles of the stromal cells were small in number, and filaments and dense attachments were found in the ectoplasm. Around the stromal cells there was a discontinuous basement membrane.     

Hypothalamic laterality in regulating gonadotropic function: unilateral hypothalamic lesion and ovarian compensatory hypertrophy

Regardless of the side of hemiovariectomy, unilateral lesion placed in the right-side medial anterior hypothalamus suppressed ovarian compensatory hypertrophy, but the lesion made in the left side failed to suppress it. This suggests the presence of a hypothalamic laterality in regulating gonadotropin secretion.     

Devastating outflow obstruction after pediatric split liver transplantation

Sakamoto S, Nakazawa A, Shigeta T, Uchida H, Kanazawa H, Fukuda A, Karaki C, Nosaka S, Kasahara M. Devastating outflow obstruction after pediatric split liver transplantation. Abstract: HVOO is a rare complication after pediatric LT, which may lead to graft failure. There are various causes of HVOO, such as mechanical anastomotic obstruction and SOS. A 10‐month‐old female underwent split LT from a deceased donor for ALF. Her postoperative course was uneventful. However, her liver function suddenly deteriorated a month later. A liver biopsy revealed centrilobular injury, and D‐US suggested outflow obstruction. Venography was performed to reveal hepatic venous narrowing inside the graft. She received another graft from a living donor because of progressive graft failure in spite of successful venoplasty with stent insertion. The macroscopic findings of the explanted graft did not show an anastomotic stricture of the hepatic vein, although the pathological findings revealed necrosis of the first graft due to SOS. SOS might cause severe consequences with concomitant mechanical outflow obstruction after pediatric LT.     

Density heterogeneity of eosinophil leucocytes: induction of hypodense eosinophils by platelet-activating factor.

We have examined the induction of hypodense eosinophils by platelet-activating factor (PAF), a mediator which may be involved in eosinophil activation in allergic diseases. Guinea-pig eosinophils were incubated with buffer or PAF and applied to continuous Percoll density gradients. Cellular density ranged from 1.0142 to 1.1369 g/ml. Peak eosinophil density in control was 1.0887 +/- 0.0008 g/ml (mean +/- SEM), and 91.1 +/- 1.4% of eosinophils were distributed between 1.0810 and 1.1000 g/ml. Preincubation of eosinophils with PAF(10(-7) M) resulted in a time-dependent and non-cytolytic increase of the number of hypodense eosinophils, with peak densities after incubation for 1 hr and 2 hr of 1.0834 +/- 0.0014 (n = 4, P less than 0.05) and 1.0755 +/- 0.0007 g/ml (n = 6, P less than 0.01), respectively. After incubation for 2 hr, 82.0 +/- 4.9% (n = 6) eosinophils showed a density lower than 1.080 g/ml. Lyso-PAF, the inactive precursor and metabolite of PAF, at a concentration of 10(-7) M had no effect on cell density. The specific PAF receptor antagonist WEB 2086 (10(-6) M) inhibited the PAF-induced density shift by 87.0 +/- 5.3%. Our results demonstrate that a single mediator is able to induce the formation of hypodense eosinophils. We conclude that the appearance of hypodense eosinophils in allergic diseases such as asthma may occur, at least in part, in response to inflammatory mediators which activate these cells.     

Changes in airway responsiveness and β- and α-adrenergic receptors in the lungs of guinea pigs with experimental asthma

The effects of inhaled allergen on airway responsiveness and on beta- and alpha-1-adrenergic receptors on lung membrane were investigated in guinea pigs. After measuring the respiratory threshold to histamine (RT-HIS), one group of guinea pigs passively sensitized for ovalbumin was challenged by allergen inhalation (challenged group). Measurement of the RT-HIS 24 h following challenge revealed a significant decrease from 687 micrograms/ml (mean, n = 16) to 407 micrograms/ml (P less than 0.05). In addition the RT-HIS 24 h after challenge was also significantly lower in the challenged group than in controls (n = 9, P less than 0.05). The density of beta-adrenergic receptors on the lung membrane of the challenged group was 594 +/- 32 (mean +/- SE) fmol/mg protein (n = 11) compared with 712 +/- 24 fmol/mg protein (n = 9) in the controls, a statistically significant difference (P less than 0.05). A significant correlation was found between the RT-HIS and density of beta-adrenergic receptors. From these results, we concluded that the exaggerated airway responsiveness 24 h after allergen challenge is in part due to a decrease in the density of beta-adrenergic receptors. There was no difference in the density of alpha-1-adrenergic receptors nor a significant correlation between the RT-HIS and the number of alpha-1-adrenergic receptors in the challenged vs. the control groups.     

The central pattern generator for vomiting may exist in the reticular area dorsomedial to the retrofacial nucleus in dogs

There is some controversy over whether or not a discrete site that integrates vomiting activities in somatic and autonomic nerves is present in the medulla oblongata. On the basis of our previous studies, we hypothesized that the temporal patterns of muscle contractions in vomiting are generated by a central pattern generator in the retrofacial area of the rostral medulla. To investigate this hypothesis further, the effects of electrical and chemical lesions of the medullary area were observed in decerebrate paralyzed dogs. Efferent activities of the phrenic and abdominal muscle nerves were recorded to recognize fictive vomiting. The right half of the medulla oblongata was transversely severed about 3 mm rostral to the obex. Fictive vomiting responses to vagal stimulation still appeared after hemisection in all 11 dogs. In addition, stimulation of the contralateral reticular area dorsomedial to the retrofacial nucleus produced fictive vomiting even after hemisection. An electrical lesion or injection of kainic acid (0.5–1.0 μl) was applied at the point where reticular stimulation induced fictive vomiting. After this destruction, no activities that corresponded to fictive vomiting could be induced by stimulation of vagal afferents or the reticular site. Salivation was decreased by hemisection, and decreased further, but was not completely abolished, with destruction of the reticular area. Kainic acid is known to selectively destroy neural cell bodies. Therefore, we concluded that neuronal somata in the reticular formation dorsomedial to the retrofacial nucleus play an essential role in the central patterning of vomiting activities in peripheral motor nerves. Received: 10 September 1996 / Accepted: 2 July 1997     

Local immune response in squamous cell carcinoma of the uterine cervix

The role of Langerhans cells as antigen-presenting cells was examined in cervical carcinomas. Frozen samples were obtained from 34 women with stage Ib and II cervical carcinomas. Langerhans cells (CD1), T lymphocytes (CD4 and CD8), B lymphocytes (CD22), and natural killer (CD57, NK) cells were all quantitatively assessed in cervical carcinomas using immunohistochemical methods. These results were related to the MHC class I and II expression on the tumor cells. The majority of Langerhans cells were distributed among cancer cells and they were positively correlated with CD4+, NK and B cells in cervical carcinomas. This is suggestive of the presence of local immune response. The numbers of Langerhans, CD4+, CD8+ and NK cells did not significantly correlate with age at operation, lymph node metastases or depth of cervical wall invasion. The downregulation of MHC class I expression found in 8 (24%) carcinomas was not associated with the decrease in the number of immunologic cells. The upregulation of MHC class II expression found in 26 (76%) carcinomas was significantly associated with the increase in the number of Langerhans cells (p < 0.007). However, the association between the upregulation of MHC-II expression and CD4+ cells did not reach statistical significance (p < 0.07). This is probably due to a small case in this study. MHC-II-restricted immunity may partly contribute to the local immune response in stages Ib and II squamous cell carcinoma of the uterine cervix.     

Fos expression induced by warming the preoptic area in rats

The preoptic area (POA) occupies a crucial position among the structures participating in thermoregulation, but we know little about its efferent projections for controlling various effector responses. In this study, we used an immunohistochemical analysis of Fos expression during local warming of the preoptic area. To avoid the effects of anesthesia or stress, which are known to elicit Fos induction in various brain regions, we used a novel thermode specifically designed for chronic warming of discrete brain structures in freely moving rats. At an ambient temperature of 22 degrees C, local POA warming increased Fos immunoreactivity in the supraoptic nucleus (SON) and the periaqueductal gray matter (PAG). Exposure of animals to an ambient temperature of 5 degrees C induced Fos immunoreactivity in the magnocellular paraventricular nucleus (mPVN) and the dorsomedial region of the hypothalamus (DMH). Concurrent warming of the POA suppressed Fos expression in these areas. These findings suggest that thermal information from the preoptic area sends excitatory signals to the SON and the PAG, and inhibitory signals to the mPVN and the DMH.     

Metabolic acidosis in patients receiving anticonvulsants

Blood pH, bicarbonate, PCO₂, serum calcium, alkaline phosphatase and red cell carbonic anhydrase were measured in 37 selected patients receiving anticonvulsants. Patients with metabolic acidosis showed a high incidence of hypocalcemia with increased alkaline phosphatase and a significant reduction of carbonic anhydrase-B activity. High iPTH levels were found in 13 patients, but this was not correlated with acid-base balance status. Anticonvulsant drugs seemed to inactive carbonic anhydrase-B activity. Metabolic acidosis might be one of the factors causing a disturbance of calcium metabolism in these patients.This research was supported by Shimabara Science Promotion Foundation, 1978