Perinatal rib fractures in 18 calves delivered from Holstein dams.

Cranial rib fractures during dystocia and the ensuing callus formations in calves often cause tracheal stenosis. Rib fractures may affect the lung since ribs tend to fracture above the costochondral junction during delivery. Considering that calving assistance rates for dystocia are high, calves with fractured ribs may develop respiratory disease which results in economic loss. The objective of this study was to elucidate the contribution of rib fractures to economic loss through respiratory disease in calves. Of 163 sick calves delivered from Holstein-Friesian dams included in this study, a total of 18 rib fractured calves was found, giving an incidence of rib fracture in sick calves of 11.0%. There were significant differences in incidence by the rib involved, indicating the 2nd to 7th ribs tend to break. Many of the rib fractured calves showed dyspnea and pyrexia. In this study, four of five scanned or necropsied calves had pneumonia lesions despite the fact that these four calves did not have tracheal stenosis. Rib fractured calves sold at below market value with a median difference from average sale price of minus 64,861 yen. Survival analysis indicated an overall association between rib fracture and time to death. In this study, we demonstrated that rib fractures happened most frequently in the 2nd to 7th ribs, and these cases tended to cause pneumonia, which decreased sale prices and longevity. Farmers should work to reduce risks and rates of dystocia so as to lessen economic loss and poor welfare in calves due to rib fractures.

Efficacy of a novel treatment serum in the improvement of photodamaged skin.

A novel treatment serum formulated to target multiple pathways in the anti-ageing cascade was tested both in vitro and in clinical settings. In vitro testing was performed to assess the ability to stimulate key proteins and genes fundamental to the anti-ageing cascade. The antioxidant potential of the formulation was studied in a UV-irradiation clinical study. A 12-week, open-label, single-centre study was conducted to determine whether this uniquely formulated topical treatment serum could improve visible signs of facial photodamage. Clinical evaluations showed statistically significant reductions in fine wrinkles and coarse wrinkles and improvements in skin texture, tone and radiance starting at week 4 with continued improvements at weeks 8 and 12. Subject self-assessments confirmed that the beneficial effects of the treatment serum were readily observed by the users. The treatment serum was well tolerated with no treatment-related adverse events reported during the 12-week study. Use of this novel treatment serum produced significant improvements in the visible signs of facial photodamage.

Effect of mixing modes on chemiluminescent detection of epinephrine with lucigenin by an FIA system fabricated on a microchip.

The chemiluminescent (CL) detection of epinephrine (EP) with lucigenin (Luc) was performed using a micro flow cell fabricated on a silicon chip. A solution of EP was injected into the Luc carrier stream. The Luc solution containing EP and an alkaline solution were successively poured into the flow cell by a pressure-driven flow system. Two types of flow cells were fabricated for estimating the effect of the mixing modes in the flow cells on the intensity of light emission. In flow cell 1, two streams entered through separate inlet ports and merged to flow adjacently. In flow cell 2, a Luc solution containing EP was split up to 36 partial flows by passage through the nozzles, and was injected into the alkaline solution. The intensity of light emission in flow cell 2 increased markedly compared to that in flow cell 1. The detection limit of 8.0 x 10(-7) M for EP in flow cell 2 was a factor of six-times better than that in flow cell 1. The improvement in the sensitivity for EP could be explained in terms of the distortion of laminar flow in flow cell 2.

Effect of the geometry of microfabricated flow reactors on chemiluminescent detection of epinephrine with lucigenin.

Three types of flow reactors with different lengths (12-126 mm) and widths (0.4-2.0 mm) of channel were made on the silicon chip by microfabrication techniques for the chemiluminescent (CL) detection of epinephrine (EP) with lucigenin (Luc). The volume of each CL reactor was about 10 microL. A solution containing EP and Luc and a solution containing NaOH and periodate were injected successively into each inlet of the CL reactor in the range 20-100 microL/min with a pressure-driven flow system. The intensity of light emission was dependent on the geometry of the flow reactors. These results could be explained in terms of the differences in the diffusion length of the reactants in the flow reactors. The maximum light emission were linearly correlated, with the concentrations of EP over the range from the detection limit of 5.0 x 10(-8) mol/L up to 5.0 x 10(-6) mol/L on the use of the CL reactor with the most promising geometry.

Identification of the suprachiasmatic nucleus in birds.

Circadian rhythms are generated by an internal biological clock. The suprachiasmatic nucleus (SCN) in the hypothalamus is known to be the dominant biological clock regulating circadian rhythms in mammals. In birds, two nuclei, the so-called medial SCN (mSCN) and the visual SCN (vSCN), have both been proposed to be the avian SCN. However, it remains an unsettled question which nuclei are homologous to the mammalian SCN. We have identified circadian clock genes in Japanese quail and demonstrated that these genes are expressed in known circadian oscillators, the pineal and the retina. Here, we report that these clock genes are expressed in the mSCN but not in the vSCN in Japanese quail, Java sparrow, chicken, and pigeon. In addition, mSCN lesions eliminated or disorganized circadian rhythms of locomotor activity under constant dim light, but did not eliminate entrainment under light-dark (LD) cycles in pigeon. However, the lesioned birds became completely arrhythmic even under LD after the pineal and the eye were removed. These results indicate that the mSCN is a circadian oscillator in birds.

Molecular analysis of avian circadian clock genes.

Unlike mammals, avian circadian rhythms are regulated by a multiple oscillatory system consisting of the retina, the pineal and the suprachiasmatic nucleus in the hypothalamus. To understand avian circadian system, we have cloned Clock and Period homologs (qClock, qPer2 and qPer3) and characterized these genes in Japanese quail. Overall, qCLOCK, qPER2 and qPER3 showed approximately 79%, approximately 46% and approximately 33% amino acid identity to mCLOCK, mPER2, mPER3, respectively. Clock was mapped to quail chromosome 4 and chicken chromosome 4q1.6-q2.1. Per2 and Per3 genes were both localized to microchromosomes. qClock mRNA was expressed throughout the day, while qPer2 and qPer3 showed robust circadian oscillation in the eye and the pineal gland. All three genes were expressed in various tissues. In addition, qPer2 mRNA was induced by light, but neither qClock nor qPer3 was induced. These results can explain the molecular basis for circadian entrainment in Japanese quail and also provide new avenues for molecular understanding of avian circadian clock and photoperiodism.

The GTPase activating factor for transducin in rod photoreceptors is the complex between RGS9 and type 5 G protein beta subunit.

Proteins of the regulators of G protein signaling (RGS) family modulate the duration of intracellular signaling by stimulating the GTPase activity of G protein alpha subunits. It has been established that the ninth member of the RGS family (RGS9) participates in accelerating the GTPase activity of the photoreceptor-specific G protein, transducin. This process is essential for timely inactivation of the phototransduction cascade during the recovery from a photoresponse. Here we report that functionally active RGS9 from vertebrate photoreceptors exists as a tight complex with the long splice variant of the G protein beta subunit (Gbeta5L). RGS9 and Gbeta5L also form a complex when coexpressed in cell culture. Our data are consistent with the recent observation that several RGS proteins, including RGS9, contain G protein gamma-subunit like domain that can mediate their association with Gbeta5 (Snow, B. E., Krumins, A. M., Brothers, G. M., Lee, S. F., Wall, M. A., Chung, S., Mangion, J., Arya, S., Gilman, A. G. & Siderovski, D. P. (1998) Proc. Natl. Acad. Sci. USA 95, 13307-13312). We report an example of such a complex whose cellular localization and function are clearly defined.

Detection of anti-type VII collagen antibody in Sjögren's syndrome/lupus erythematosus overlap syndrome with transient bullous systemic lupus erythematosus.

Bullous systemic lupus erythematosus (SLE) is a chronic, widespread, non-scarring, subepidermal blistering eruption associated with autoimmunity to type VII collagen. We describe a patient with Sj ogren's syndrome/lupus erythematosus overlap syndrome who showed transient blistering eruptions over limited skin surface and in oral mucosa. At the time of aggravation, the patient's serum contained IgG autoantibodies that bound to the dermal side of 1 mol/L NaCl-split normal skin, as determined by an indirect immunofluorescence test, and that reacted to type VII collagen, as determined by immunoblotting on dermal extract. Our observations suggest that a chronic, widespread, blistering eruption is not a prerequisite for the diagnosis of bullous SLE, and a mild, transient, blistering eruption could be a manifestation of type I bullous SLE.

Neutrophilic eccrine hidradenitis: report of two cases.

BACKGROUND: Neutrophilic eccrine hidradenitis (NEH) is an uncommon, self-limited dermatosis usually attributed to anti-cancer chemotherapy. It is characterized histologically by necrosis of the eccrine gland and neutrophilic infiltrate. OBSERVATIONS: We saw NEH in a 5-year-old boy with acute lymphoblastic leukemia and a 4-year-old girl with acute monocytic leukemia. NEH developed after the anti-leukemic chemotherapy including high dose cytarabine. The eruption was composed of vesicles, papules, and plaques. CONCLUSIONS: Histological findings were compatible with those described in the literature. NEH in our two patients could be attributed to high doses of cytarabine.

Studies on antiallergic agents. II. Quantitative structure-activity relationships of novel 6-substituted N-(1H-tetrazol-5-yl)-2-pyrazinecarboxamides.

The effect of structural modifications of 6-substituted N-(1H-tetrazol-5-yl)-2-pyrazinecarboxamides on their anti-allergic activity was analyzed quantitatively by means of the Hansch-Fujita method. The activity of these compounds was correlated with hydrophobic (pi) and steric (molecular refractivity and STERIMOL B1) effects of the 6-substituent on the pyrazine ring. The 6-substituents with a length greater than n-propylamino possess an extra effect enhancing the activity. Moreover, the activity increased progressively from 6-non-amino via alkylamino- to dialkylamino-substituted compounds, other factors being equal. This could be attributable to an electronic effect of substituents. Electron-donating small and yet symmetric substituents with high hydrophobicity longer than n-propylamino seemed to be favorable to the activity. By compromising these contradictory requirements, small dialkylamino (including cyclic amino) groups were decided to be the most favorable substituents. This analysis was in agreement with the observation that the most effective compounds were the 6-dimethylamino (I-27) and 6-(1-pyrrolidinyl) (I-34) derivatives.

Inhibitory effect of HSR-6071, a new anti-allergic agent, on experimental asthma in rats and guinea-pigs.

The experimental asthma caused by IgE antibody in rats was inhibited by HSR-6071 (6-(1-pyrrolidinyl)-N-(1H-tetrazol-5-yl)-2-pyrazinecarboxamide) (0.01-0.1 mg kg-1 i.v.) in a dose-dependent manner. The inhibitory activity of HSR-6071 was more potent than those of disodium cromoglycate and ketotifen, and equipotent with amlexanox. The bronchoconstriction mediated by IgE or IgG antibody in guinea-pigs was also prevented by HSR-6071 (0.3, 1 and 3 mg kg-1 i.v.), amlexanox (3, 10 and 30 mg kg-1 i.v.) and ketotifen (0.1 mg kg-1 i.v.) but not by disodium cromoglycate (10 mg kg-1 i.v.). HSR-6071 was more potent than amlexanox, but less potent than ketotifen. HSR-6071 suppressed antigen-induced histamine and SRS-A release from minced lung tissues of guinea-pigs sensitized passively with rabbit anti-EA serum and was a more potent inhibitor of the release of SRS-A than of histamine. On the other hand, histamine- or acetylcholine-induced bronchoconstriction in guinea-pigs was scarcely affected by HSR-6071 at doses sufficient to inhibit the experimental asthma, but LTD4-induced bronchoconstriction was dramatically inhibited. These results indicate that the inhibitory action on experimental allergic asthma of HSR-6071 may be due to suppression of antigen-induced histamine and SRS-A release from lung tissues and to antagonism of SRS-A action. In addition, HSR-6071 inhibited cyclic AMP phosphodiesterase activity and produced relaxation of the guinea-pig isolated trachea. These pharmacological actions may contribute to the production of the anti-allergic action of HSR-6071.

Studies on antiallergic agents. I. Synthesis and antiallergic activity of novel pyrazine derivatives.

Various pyrazine derivatives were synthesized and their antiallergic activity was examined. The inhibitory activity on allergic histamine release of the compounds bearing a 5-tetrazolyl group was more potent than that of the corresponding carboxyl derivatives. The introduction of -CONH- or -NHCO- between the pyrazine ring and the 5-tetrazolyl group as a spacer greatly enhanced the activity. N-(1H-Tetrazol-5-yl)-2-pyrazinecarboxamide (I-3) was estimated to exhibit nearly the same potency as disodium cromoglycate (DSCG). The structure-activity relationship among various derivatives modified by introducing some substituents onto the 3-, 5- or 6-position of the pyrazine ring of I-3 was investigated. The activity remained unchanged or was reduced when such substituents as methyl, chloro, methoxy, methylamino and dimethylamino were introduced at the 3- or 5-position. In contrast, 6-substitution with various alkylamino groups more or less increased the activity. Among them, the 6-dimethylamino (I-17c) and 6-(1-pyrrolidinyl) (I-34) derivative were proved to be most potent. The IC50 values (concentration which produces 50% inhibition of the allergic histamine release) of I-17c and I-34 were determined to be 4.7 x 10(-10) and 4.6 x 10(-10) M, respectively. These two compounds produced a potent inhibitory activity on passive cutaneous anaphylaxis (PCA) in rat, not only by the intravenous route (ED50 = 0.0096 mg/kg for both compounds) but also by the oral route (ED50 = 0.19 and 0.18 mg/kg, respectively). On the other hand, when the pyrazine ring of some representative compounds was replaced with a pyridine ring, the inhibitory activity on histamine release was significantly reduced.

Potent inhibitory activity of HSR-6071, a new antiallergic agent, on passive cutaneous anaphylaxis (PCA).

Antiallergic effects of 6-(1-pyrrolidinyl)-N-(1H-tetrazol-5-yl)-2-pyrazinecarboxamide (HSR-6071), a newly synthesized agent, were investigated. The 48-hr homologous passive cutaneous anaphylaxis (PCA) in rats was inhibited in a dose-dependent manner by i.v. and p.o. administration of the agent (ED50 = 0.0096 mg/kg and 0.18 mg/kg, respectively). The IgE-mediated histamine release from rat peritoneal exudate cells was inhibited by HSR-6071, with an IC50 of 4.6 x 10(-10) M. Regarding the non-immunological histamine release, HSR-6071 inhibited compound 48/80-induced, but not A23187-induced and spontaneous histamine release. On the other hand, an increase in vascular permeability induced by histamine, serotonin and bradykinin was unaffected by HSR-6071 in doses sufficient to inhibit PCA. In addition, the contractile responses of isolated guinea pig ileum to histamine, acetylcholine and serotonin were also unaffected by the agent even in a high concentration of 10(-4) M. These results indicate that HSR-6071 possesses a potent antiallergic activity and that the inhibition of PCA by HSR-6071 may be due to the suppression of chemical mediators release from mast cells.

Histopathologic study on tumor infiltration into the intervertebral disc.

This study was performed in order to understand the reasons why the metastasis of malignancy into the disc is rare in contrast to the commonly seen vertebral metastasis. By histopathologic studies of 14 vertebral columns with vertebral metastases of various malignant tumors and 616 disc specimens, the authors found three pathways for an intravertebral tumor to invade into the adjacent disc: 1) direct infiltration from the rim of the vertebral body not covered by the cartilagenous plate, 2) infiltration from the side of the vertebral body close to the vertebral corner, through the subspace beneath the longitudinal ligament, and 3) hematogenous invasion via small vessels in the subspace beneath the longitudinal ligament. The authors concluded that the cartilagenous plate between the vertebral body and the disc as well as high intradiscal pressure would be acting as a barrier against tumor invasion from the vertebral body into the disc, and suggests that gradual increase of capillaries in the disc with age may enhance hematogenous invasion into the disc in rare occasions.

Diabetes mellitus in Kearns-Sayre syndrome.

A 20-year-old woman with Kearns-Sayre syndrome (KSS) suddenly experienced two episodes of diabetic coma. She was studied to determine whether diabetes mellitus (DM) resulted from insulin resistance or from an insulin secretion abnormality, using the euglycemic glucose clamp technique and the glucagon tolerance test. She had a deficiency of insulin secretion from beta cells. It is important to recognize in practice the onset of DM in patients with mitochondrial myopathy. We would suggest that a genetic linkage or mitochondrial dysfunction may be responsible for the association of both disease states.