Human brain natriuretic peptide-like immunoreactivity in human brain.

The presence of immunoreactive human brain natriuretic peptide in the human brain was studied with a specific radioimmunoassay for human brain natriuretic peptide-32. This assay showed no significant cross-reaction with human alpha atrial natriuretic peptide, porcine brain natriuretic peptide or rat brain natriuretic peptide. Immunoreactive human brain natriuretic peptide was found in all 5 regions of human brain examined (cerebral cortex, thalamus, cerebellum, pons and hypothalamus) (0.6-6.7 pmol/g wet weight, n = 3). These values were comparable to the concentrations of immunoreactive alpha atrial natriuretic peptide in human brain (0.5-10.1 pmol/g wet weight). However, Sephadex G-50 column chromatography showed that the immunoreactive human brain natriuretic peptide in the human brain eluted earlier than synthetic human brain natriuretic peptide-32. These findings suggest that human brain natriuretic peptide is present in the human brain mainly as larger molecular weight forms.     

Calcitonin gene-related peptide-like immunoreactivity in the human small intestine.

Calcitonin-gene-related-peptide (CGRP)-like immunoreactivity was localized in nerve fibres, neuronal somata and in mucosal endocrine cells of the human small intestine. Immunoreactive enteric neurons were more numerous in the submucous plexuses than in the myenteric plexus. Morphologically, they predominantly had the appearance of type II neurons. The majority of the CGRP-like immunoreactive nerve fibres ran within the ganglionic nerve plexuses. Only a small proportion could be observed in the lamina propria, the lamina muscularis mucosae, or the circular and longitudinal outer smooth muscle layer. These findings suggest that within the wall of the human small intestine neuronal CGRP of either extrinsic or intrinsic origin exerts its effect chiefly on other enteric neurons, and might be indirectly involved in the regulatory functions of the human small intestine.     

The distribution of galanin message-associated peptide-like immunoreactivity in the pig.

Using a radioimmunoassay (RIA) developed to the N-terminal part of the predicted sequence of porcine galanin message-associated peptide (GMAP), we have confirmed the existence of GMAP-like immunoreactivity (-LI) in normal porcine tissues. GMAP-LI was found to parallel the distribution of galanin-immunoreactivity (-IR), although consistently the concentrations detected were, on a molar ratio, significantly less than those measured for galanin throughout the gastrointestinal tract, brain, spinal cord, adrenal and pituitary gland. As cleavage of the prohormone would be expected to produce galanin and GMAP on an equimolar basis, it is possible that the endogenous, intact GMAP peptide does not fully cross-react with the antibody raised to the N-terminal GMAP sequence. Gel chromatography of tissue extracts revealed a single molecular form of galanin-IR in the gut and four distinct molecular forms in the adrenal gland. GMAP-LI eluted as a single immunoreactive component in the gut, and in the adrenal gland there were two major molecular forms, one of which was apparently also detected by the galanin assay, and a small amount of N-terminal fragment. This molecular heterogeneity seems likely to be a result of the various possible prohormone cleavage products and/or posttranslational processing modifications. Further analysis of the galanin gene products needs to be undertaken in order to confirm this.     

Leptin receptor in the chicken ovary: potential involvement in ovarian dysfunction of ad libitum-fed broiler breeder hens

In hens, the ovarian follicles committed to ovulation are arranged in an ordered follicular hierarchy. In standard broiler breeders hens genetically selected for high growth rate the reproductive function is clearly dysfunctional. Feed restriction is needed during reproductive development to limit the formation of excessive numbers of ovarian yellow follicles arranged in multiple hierarchies. To determine whether leptin is involved in the nutritional and reproductive interactions controlling follicular hierarchy in hens, blood leptin levels and ovarian expression of the leptin receptor mRNA were determined during follicle maturation in three chicken lines; a slow growing broiler "Label" genotype without reproductive dysfunction, a fast growing "Standard" genotype fed ad libitum or restricted and a fast growing "Experimental" line with intermediate reproductive performance levels. Whereas expression of the leptin receptor mRNA did not change in the theca, it clearly decreased with follicular differentiation in the granulosa of slow growing hens. In fast growing standard hens fed ad libitum and presenting significant reproductive dysfunction, the decrease was disrupted and dramatic up-regulation of granulosa cell expression of the leptin receptor was observed. On the other hand, feed restriction decreased the overall level of expression of the leptin receptor mRNA and restored the decrease with follicular growth. The level of expression of the leptin receptor probably modulates the action of leptin on follicular differentiation. Since blood leptin and other metabolic factors were not affected by the genotype or by nutritional state, the factors involved in the regulation of leptin receptor gene expression remain to be determined. This study demonstrates the involvement of leptin in the nutritional control of reproduction in birds. Leptin action on the ovary probably controls follicular hierarchy through the regulation of steroidogenesis.     

Calcitonin gene-related peptide-like immunoreactivity in nerve fibers in the human skin. Relation to fibers containing substance P-, somatostatin- and vasocactive intestinalpolypeptide-like immunoreactivity

Calcitonin gene-related peptide-like immunoreactivity was demonstrated in in sensory nerve fibers in the epidermis and dermis as free nerve endings and around blood vessels and hair follicles of the human finger pad and arm skin. The vast majority of the calcitonin gene-related immunoreactive fibers was shown to display also substance P-like immunoreactivity and a few fibers in the dermis were somatostatin positive. No fibers displaying both substance P and somatostatin-like immunoreactivity were found but a few substance P immunoreactive fibers in the dermis-epidermis region were found to contain also vasointestinal polypeptide-like immunoreactivity. In the sweat glands, abundant calcitonin gene-related peptide positive, but substance P negative, fibers were observed with a similar distribution pattern as the vasoactive intestinal polypeptide immunoreactive fibers and these fibers were suggested to be of sympathetic origin.     

Characterization of human thyrotropin receptor-related peptide-like immunoreactivity in peripheral blood of Graves' disease.

An antiserum raised against an alignment of amino acid-(32-56), termed TSHRP-1, in the extracellular domain of human thyrotropin (TSH) receptor was used to identify the TSH receptor-like substance in plasma of Graves' disease. The dilution curve of plasma TSHRP-1-like immunoreactivity was observed in a manner parallel to the standard synthetic peptide curve in radioimmunoassay, and its molecular weight estimated approximately 60 kDa. The amounts of TSHRP-1-like immunoreactivity were significantly higher in Graves' plasma than those in plasma of normal and hypothyroid patients due to Hashimoto's thyroiditis. The present results indicate that human peripheral blood possesses a soluble form of the extracellular domain of TSH receptor which may contribute to the pathophysiology of Graves' disease.     

Atrial natriuretic peptide-like immunoreactivity in neurons and astrocytes of human cerebellum and inferior olivary complex.

Atrial natriuretic peptide (ANP) has previously been localized in areas of mammalian brain associated with olfaction, cardiovascular function, and fluid/electrolyte homeostasis. Despite the presence of several types of natriuretic peptide receptors in mammalian cerebellum, neither intrinsic nor extrinsic sources of the natriuretic peptides have been described. In this report we describe the immunohistochemical localization of both intrinsic and extrinsic sources for ANP in human cerebellum. ANP-like immunoreactivity (ANP-LIR) was observed in climbing fibers in the cerebellar molecular layer that probably originated from isolated immunopositive neurons of the inferior olivary complex. Intrinsic sources of ANP-LIR included small subpopulations of protoplasmic and fibrous astrocytes and Bergmann glia, as well as Golgi and Lugaro neurons of the granule cell layer. These results suggest that, in addition to its presumptive roles in local vasoregulation, ANP may serve as a modulator of the activity of Purkinje neurons.     

Effect of dietary aflatoxin on fertility, hatchability, and progeny performance of broiler breeder hens.

The effects of aflatoxin on egg production, fertility, hatchability, and progeny performance were investigated by feeding dietary aflatoxin at dose levels of 0,5, and 10 mug/g to mature broiler breeder hens for 4 weeks. Sixteen hens were used for each dietary dose level. Egg production decreased significantly during weeks 3 and 4 after initiation of toxin feeding for hens fed 10 and 5 mum of aflatoxin per g of diet respectively. Whereas fertility was not affected by dietary aflatoxin, hatchability of fertile eggs decreased significantly within week 1 of toxin feeding. Hatchability of fertile eggs collected during week 1 of the treatment period was 95.1, 68.9, and 48.5% for the control, 5- and 10-mug/g groups, respectively. At the dose levels used in this study, no latent effects of the aflatoxin or its metabolites were observed on the performance of surviving chicks. Six hens from each experimental groups were necropsied at the end of the 4-week treatment period. These birds exhibited typical symptoms of aflatoxicosis, including enlarged, fatty and friable livers, and enlarged spleens.     

Effect of dietary aflatoxin on fertility, hatchability, and progeny performance of broiler breeder hens.

The effects of aflatoxin on egg production, fertility, hatchability, and progeny performance were investigated by feeding dietary aflatoxin at dose levels of 0,5, and 10 mug/g to mature broiler breeder hens for 4 weeks. Sixteen hens were used for each dietary dose level. Egg production decreased significantly during weeks 3 and 4 after initiation of toxin feeding for hens fed 10 and 5 mum of aflatoxin per g of diet respectively. Whereas fertility was not affected by dietary aflatoxin, hatchability of fertile eggs decreased significantly within week 1 of toxin feeding. Hatchability of fertile eggs collected during week 1 of the treatment period was 95.1, 68.9, and 48.5% for the control, 5- and 10-mug/g groups, respectively. At the dose levels used in this study, no latent effects of the aflatoxin or its metabolites were observed on the performance of surviving chicks. Six hens from each experimental groups were necropsied at the end of the 4-week treatment period. These birds exhibited typical symptoms of aflatoxicosis, including enlarged, fatty and friable livers, and enlarged spleens.     

Atrial natriuretic peptide-like immunoreactivity in astrocytes of parenchyma and glia limitans of the canine brain.

The Na(+)-regulating and vasoactive peptide hormone atrial natriuretic peptide (ANP) has been localized in cultured glial cells from neonatal rat brain but has not been reported in situ in adult brain. In the present study, immunohistochemical localization of ANP in astrocytes of adult canine brain was obtained using an antibody against rat ANF IV, a 25-amino acid fragment identical to low molecular weight brain ANP. ANP-like immunoreactivity was localized in widely dispersed astroglia throughout the canine brain, including Types I and II, in addition to some astrocytes of the glia limitans (GL).     

Dendroaspis natriuretic peptide-like immunoreactivity and its regulation in rat aortic vascular smooth muscle

Dendroaspis natriuretic peptide (DNP) is a recently isolated 38 amino acid peptide that shares structural and functional properties with the other members of the natriuretic peptide family. The present study demonstrates the presence of DNP-like immunoreactivity in sections of rat aorta, carotid artery and renal vasculature and tubules. DNP-like immunoreactivity was detected in culture aortic vascular smooth muscle cells and medium and is regulated by endothelin-1, angiotensin II and sodium nitroprusside but not by transforming growth factor-beta. Our observations indicate that DNP elicits a marked inhibitory effect on DNA synthesis in culture rat aortic vascular smooth muscle cells.     

Calcitonin gene-related peptide-like immunoreactivity in the central nervous system and peripheral organs of rats

The concentrations of rat calcitonin gene-related peptide-like immunoreactivity (rCGRP-LI) in various organs of male rats as well as the molecular heterogeneity of rCGRP-LI in tissue extracts was examined using a specific radioimmunoassay (RIA) for rCGRP and gel-filtration chromatography. rCGRP-LI was high in extracts of the spinal cord (202 +/- 22.6 pg/mg wet wt. of tissue; mean +/- S.E.M.) and of the thyroid (229 +/- 62.3 pg/mg). rCGRP-LI was detectable in the brainstem, hypothalamus, stomach, duedenum, pancreas and kidney. The elution pattern of the extracts on a Sephadex G-50 column showed 3 peaks of rCGRP-LI irrespective of organs and tissues. The first peak corresponded to authentic rCGRP-(1-37). The second and third rCGRP-LI peaks probably consisted of C-terminal fragments of rCGRP, because they had a lower molecular weight than rCGRP-(1-37) and because our antiserum cross-reacts with a synthetic C-terminal fragment. The ratio of 3 rCGRP-LI molecules, however, differed between neural tissue extracts and others. The main component of rCGRP-LI in neural tissue was authentic rCGRP-(1-37), while the smaller fragments of rCGRP were chiefly contained in other tissues like the stomach, pancreas and thyroid. The relative ratio of rCGRP-LI molecules with different size in respective tissue extracts was not changed after leaving the dissected tissues for 2 h at room temperature. These findings indicate that rCGRP-LI is abundantly present in the thyroid as well as the spinal cord and it is detected in lower amounts in the alimentary tract and central nervous system. rCGRP-LI in the extracts consists of 3 different components, the proportions of which vary from one tissue to another, probably reflecting tissue-specific differences in the processing of CGRP.     

Small cardioactive peptide-like immunoreactivity and its colocalization with FMRFamide-like immunoreactivity in the central nervous system of the leech Hirudo medicinalis

Summary The distributions of small cardioactive peptide (SCP)- and FMRFamide-like immunoreactivities in the central nervous system of the medicinal leech Hirudo medicinalis were studied. A subset of neurons in the segmental ganglia and brains was immunoreactive to an antibody directed against SCP_B. Immunoreactive cell bodies were regionally distributed throughout the nerve cord, and occurred both as bilaterally paired and unpaired neurons. The majority of the unpaired cells displayed a tendency to alternate from side to side in adjacent ganglia. A small number of neurons were immunoreactive only in a minority of nerve cords investigated. Intracellular injections of Lucifer yellow dye and subsequent processing for immunocytochemistry revealed SCP-like immunoreactivity in heart modulatory neurons but not in heart motor neurons. FMRFamide-like immunoreactivity was also detected in cell bodies throughout the central nervous system. A subset of neurons contained both SCP- and FMRFamide-like immunoreactivities; others stained for only one or the other antigen. These data suggest that an antigen distinct from FMRFamide is responsible for at least part of the SCP-like immunoreactivity. This antigen likely bears some homology to the carboxyl terminal of SCP_A and SCP_B.     

Distribution of vasoactive intestinal peptide-like immunoreactivity in the taste organs of teleost fish and frog

Summary Using immunohistochemistry, vasoactive intestinal peptide (VIP) was visualized in taste bud cells of the carp, Cyprinus carpio, and the European catfish, Silurus glanis, by means of light and electron microscopy. Intracellular membrane systems, presumably smooth endoplasmic reticulum, of light (sensory) cells, but not of dark (supporting) cells and basal cells, were densely labelled with antibody. In the frog (four species: Rana temporaria, R. ridibunda, R. arvalis, R. pipiens), taste bud cells did not label. However, the dense basal nerve fibre plexus, some subepithelial ganglionic cells, but no ascending intragemmal fibres, were immunoreactive. In fish, the results support evidence that VIP is involved in the modulation of taste transduction at the level of receptor cells. In the frog, an indirect, possibly vasodilatatory effect on taste perception may be considered.     

Cysteamine induced-duodenal ulcers are associated with a selective depletion in gastric and duodenal calcitonin gene-related peptide-like immunoreactivity in rats.

We have measured the endogenous levels of gastric and duodenal calcitonin gene-related peptide (CGRP)-, neurokinin A (NKA)-, galanin-vasoactive intestinal polypeptide (VIP)- and neuropeptide Y (NPY)-like immunoreactivity (li) in relation to cysteamine-induced gastric lesions and duodenal ulcers in rats. CGRP-li but not NKA-, galanin-, VIP- or NPY-li was decreased in gastric and duodenal samples following a single ulcerogenic dose of cysteamine (900 mg/kg p.o.). Temporal relationships of this phenomenon showed that CGRP-li was selectively decreased (stomach 45%, duodenum 68% as compared to controls, respectively after 24 h) concomitantly to the formation of acute gastric lesions and duodenal ulcers. Animals bearing healed ulcers 12 days after cysteamine, had gastroduodenal CGRP-li similar to control values. Pretreatment with the selective sensory neurotoxin capsaicin decreased gastroduodenal CGRP-li but not NKA-, galanin-, VIP- or NPY-li, showing that CGRP might be considered a marker of the afferent innervation of the gastroduodenal tract. The residual gastroduodenal CGRP-li levels in capsaicin-pretreated animals were not decreased by cysteamine administration, indicating that the effect of cysteamine is restricted to a peptide pool of primary afferent origin. Duodenal CGRP-li is selectively decreased by the duodenal ulcerogen cysteamine during the acute phase of ulcers formation and might be among the local mediators which afford protection against the ulcerogenic stimuli.     

MX1 exon 13 polymorphisms in broiler breeder chickens and associations with commercial traits

The Mx1 gene in mice is induced by type I interferons and is the major determinant of resistance to influenza virus and related orthomyxoviruses. It has been previously shown that a SNP in exon 13 of the chicken MX1 gene determines differential antiviral activity of the protein. We evaluated this SNP and two additional SNPs in elite broiler lines by PCR amplification and sequence analysis. Associations between MX1 exon 13 SNPs and several traits of economic interest were evaluated. Significant associations were found between the SNP determining antiviral activity and mortality in one line and leg defects in another line.     

Effects of oxygen concentration during incubation and broiler breeder age on embryonic heat production,chicken development,and 7-day performance

Older breeder flocks produce eggs with a relatively larger yolk and thereby a higher nutrient availability than young breeder flocks. To optimise nutrient utilisation and embryonic development throughout incubation and posthatch period, embryos originating from older breeder flocks may require a higher oxygen availability. The current study investigated effects of broiler breeder flock age and incubational oxygen concentration on embryonic metabolism and chicken development until 7-day posthatch. Similar sized eggs of a young (28–32 week) or old (55–59 week) Cobb 500 breeder flock were incubated at one of three oxygen concentrations (17%, 21% or 25%) from day 7 of incubation until 6 h after emergence from the eggshell. Posthatch, chickens were reared until 7 days of age. Egg composition at the start of incubation, heat production during incubation, and embryo or chicken development at embryonic day (ED)14 and ED18 of incubation, 6 h after hatch and day 7 posthatch were evaluated. An interaction was found between breeder age and oxygen concentration for yolk-free body mass (YFBM) at ED18. A higher oxygen concentration increased YFBM in the old breeder flock, whereas no difference was found between 21 and 25% oxygen in the young breeder flock. Yolk size was larger in the old compared to the young flock from ED0 until 6 h after hatch. Breeder flock age did not affect YFBM at ED14 and 6 h after hatch nor daily embryonic heat production, but there were some effects on relative organ weights. Chickens of the old compared to the young breeder flock showed a higher weight gain at day 7, but at a similar feed conversion ratio (FCR). A higher oxygen concentration during incubation stimulated embryonic development, especially between 17% and 21% of oxygen, in both flock ages_. Although this growth advantage disappeared at 7 days posthatch, a low oxygen concentration during incubation resulted in a higher FCR at 7 days posthatch. Results indicated that breeder flock age seemed to influence body development, with an advantage for the older breeder flock during the posthatch period. Oxygen concentrations during incubation affected body development during incubation and FCR in the first 7 days posthatch. Although an interaction was found between breeder flock age and oxygen concentration at ED18 of incubation, there was no strong evidence that nutrient availability at the start of incubation (represented by breeder flock ages) affected embryo and chicken development at a higher oxygen concentration.     

Effects of broiler breeder strain,age, and eggs preheating profile in single-stage systems on the hatchability of eggs and quality of chicks

Preheating can increase the efficiency of commercial broiler breeding. A slow increase in temperature can counteract the negative effects of water condensation on the surface of the shell and reduces the thermal shock to the embryos. It is essential to design different preheating profiles because of the variances between breeds in terms of egg construction and the course of embryogenesis, along with the influence of the parental flocks' age on hatching rates. This study aimed to analyze six preheating profiles with different time and temperature patterns (A–F), which were immediately applied before the egg incubation began. These profiles were used for two broiler breeder strains – Hubbard Flex and Ross 308 – for the entire laying cycle divided into six age ranges: 25–30, 31–36, 37–42, 43–48, 49–54, and 55–60 wk of life. A total of 7 839 250 Hubbard Flex and 57 167 060 Ross 308 eggs were used in the study. The eggs were stored for 4–7 d before the preheating profile was applied. Mortality during incubation, hatchability, and the quality of chicks were analyzed. The most favorable results were obtained with profile B – 02 h (25.0 °C):02 h (29.4 °C):02 h (32.2 °C):02 h (35.0 °C):02 h (37.8 °C), while the worst results were obtained with profile C – 03 h (23.9 °C):03 h (25.0 °C):04 h (32.2 °C). These findings indicated that compared to a rapid temperature increase during incubation, multistage slow heating of eggs is more advantageous as it enables achieving better hatchability from apparent fertile eggs and obtaining more first-grade chicks. An analysis of the interaction of different preheating profiles with flock age and breed of parental flock revealed that profile C is not suitable for eggs from older flocks, in particular Ross 308. Deterioration of hatchability of chicks from apparent fertile eggs was also confirmed for flocks over 49 wk of life after the application of profile E – 03 h (77 °C):02 h (29.4 °C):02 h (32.2 °C):01 h (35.0 °C):02 h (37.8 °C). Increased early and late mortality of embryos was observed in the case of Hubbard Flex after the use of profile D – 03 h (25.0 °C):02 h (29.4 °C):01 h (32.2 °C):02 h (35.05 °C):02 h (37.8 °C). Based on the obtained results, it can be recommended that profile B can be applied for the entire duration of Ross 308 and Hubbard Flex flock utilization to achieve high egg hatchability and obtain high-quality chicks.     

苏北地区白羽肉鸡父母代种鸡及其商品鸡中沙门氏菌的分离、鉴定与生物特性

【背景】沙门氏菌是一种重要的人兽共患食源性致病菌,控制沙门氏菌在动物中的感染和在食品生产链中的污染对于畜牧生产、公共卫生和食品安全具有重要意义。【目的】调查苏北地区某白羽肉鸡父母代种鸡及其商品鸡的沙门氏菌感染和流行情况,为其沙门氏菌防治提供参考依据。【方法】于2018-2019年从江苏泰州、宿迁、盐城、徐州、连云港等地区分别采集某白羽肉鸡父母代种鸡场产蛋鸡及其商品鸡场1-2日龄弱雏和10日龄雏鸡的泄殖腔拭子样品共360份,进行沙门氏菌的分离与鉴定,用纸片法和PCR方法对菌株耐药表型及相关耐药基因进行检测,通过多位点序列分型(multilocussequencetyping,MLST)技术对分离菌株的亲缘关系进行研究。【结果】从120份父母代产蛋鸡分离到3株沙门氏菌,从120份商品代1-2日龄弱雏分离到25株沙门氏菌,从120份商品代10日龄雏鸡分离到82株沙门氏菌。分离的沙门氏菌对多粘菌素B、亚胺培南、多西环素、甲氧苄啶、替加环素、氟苯尼考、头孢噻肟、恩诺沙星表现为全部敏感,对呋喃妥因、青霉素、利福平、利奈唑胺、万古霉素均表现为全部耐药;对阿莫西林耐药的菌株有30株(占27.27%),...