Deleterious effects of perinatal exposure to potassium bromate on the development of offspring of Swiss mice.

The present study aimed to investigate the impact of perinatal potassium bromate (KBrO3) exposure on the development of sensorimotor reflexes and redox status, and on the histological architecture of the brain, liver, and kidney of newborn mice. Pregnant mice received 1-ml bottled drinking water daily by oral intubation and served as the control group. Another group of pregnant mice were supplemented orally with 200 mg/kg body weight KBrO3 dissolved in drinking water from gestation day 5 to postnatal day 21. KBrO3 induced a decrease in the postnatal body weight in the newborn mice. KBrO3-exposed newborn mice showed poor performance and delayed development of the sensorimotor reflexes. Histological changes, increased lipid peroxidation, and altered antioxidants were reported in the cerebrum, cerebellum, medulla oblongata, liver, and kidney of the KBrO3-exposed newborn mice. In conclusion, these findings demonstrated that perinatal exposure to bromate induced oxidative stress, histological and behavioral alterations, and was a potential teratogen in newborn mice.

Segmental spinal dysgenesis with caudal agenesis in a Holstein calf.

A rare complex dysraphic malformation, comprising segmental spinal dysgenesis with caudal agenesis, was found in a Holstein calf that was unable to stand and was slightly short at the lumbosacral spine with taillessness. The thoracolumbar and sacrococcygeal regions of the midline axial segments showed severe deformities. In the spinal cord, the thoracolumbar region showed severe constriction with myelodysplastic changes, and the sacrococcygeal region showed dorsoventral separation with connection to a neural mass. In the spine, vertebral anomalies according to the degree of the segmentation error were confirmed. The cervical and thoracic segments also showed milder dysraphic changes. These changes suggest a multisegmental causal insult impairing the early embryonic notochord. This represents the first bovine case definitively confirmed morphologically.

Targeted inactivation of dipeptidyl peptidase 9 enzymatic activity causes mouse neonate lethality.

Dipeptidyl Peptidase (DPP) 4 and related dipeptidyl peptidases are emerging as current and potential therapeutic targets. DPP9 is an intracellular protease that is regulated by redox status and by SUMO1. DPP9 can influence antigen processing, epidermal growth factor (EGF)-mediated signaling and tumor biology. We made the first gene knock-in (gki) mouse with a serine to alanine point mutation at the DPP9 active site (S729A). Weaned heterozygote DPP9 (wt/S729A) pups from 110 intercrosses were indistinguishable from wild-type littermates. No homozygote DPP9 (S729A/S729A) weaned mice were detected. DPP9 (S729A/S729A) homozygote embryos, which were morphologically indistinguishable from their wild-type littermate embryos at embryonic day (ED) 12.5 to ED 17.5, were born live but these neonates died within 8 to 24 hours of birth. All neonates suckled and contained milk spots and were of similar body weight. No gender differences were seen. No histological or DPP9 immunostaining pattern differences were seen between genotypes in embryos and neonates. Mouse embryonic fibroblasts (MEFs) from DPP9 (S729A/S729A) ED13.5 embryos and neonate DPP9 (S729A/S729A) mouse livers collected within 6 hours after birth had levels of DPP9 protein and DPP9-related proteases that were similar to wild-type but had less DPP9/DPP8-derived activity. These data confirmed the absence of DPP9 enzymatic activity due to the presence of the serine to alanine mutation and no compensation from related proteases. These novel findings suggest that DPP9 enzymatic activity is essential for early neonatal survival in mice.

Pre-clinical validation of a vaginal cream containing copaiba oil (reproductive toxicology study).

The aims of this study was to evaluate the effects of oil-resin of Copaiba (Copaifera duckei Dwyer), aired in vaginal cream on the reproductive performance of female rats (Rattus norvegicus). To determine the components of the C. duckei oleoresin, gas chromatography coupled with mass spectrometry (CG-MS) was used, and considering the trans-caryophyllene sesquiterpene as a phytochemical marker in the oleoresin. Due to the extensive use of copaiba oleoresin in the suppository form for gynecological infections, an evaluation was carried out on the effects of copaiba oleoresin (Copaifera duckei Dwyer), delivered in a vaginal cream, on the reproductive performance of female Wistar rats. For this purpose, three groups (n=5-6/group) of female rats were treated as follows: 1--vaginal cream of copaiba oleoresin (28.6 mg/kg), 2--base vaginal cream and 3--control (physiological saline 0.9%), administered intravaginally, for 30 days before pregnancy, and from day zero to day 20 during pregnancy. Laparotomy was performed on the 21st day of pregnancy, followed by the determination of reproductive variables: number of live and dead fetuses, mass of the fetuses and placentas, number of implantations and resorptions, number of corpora lutea, pre- and post-implantation loss, and analyses of the fetuses with regard to external and internal anomalies and/or malformations (skeletal and visceral). The trans-caryophyllene present in the sample is suggested as a phytochemical marker and the results of this study demonstrate an absence of maternal toxicity and foetotoxicity embryofoetotoxicity at the dose administered, corresponding to ten times the recommended dose for use in humans. Accordingly, no significant statistical difference was observed between the treated and control groups, for the variables analyzed. Thus, it is concluded that the vaginal cream containing 2.5% copaiba oleoresin is safe during gestation, in female rats (Rattus norvegicus) of the Wistar strain.

Detection of chicken anemia virus and infectious bursal disease virus co-infection in broilers / Detecção do virus da anemia das galinhas em coinfecção com o vírus doença infecciosa bursal em frangos

This survey aimed to investigate chicken anemia virus (CAV) in broilers flocks experimenting retarded growth and increasing mortality since the fourth day of age. Clinically, chickens presented depression, paleness, depigmentation and retarded growth. At necropsy, chickens presented CAV-compatible lesions. Samples from liver, spleen and thymus were tested by PCR for a 675-bp fragment of the CAV VP-1 gene, and all tested samples were positive. Serological and molecular techniques did not detect other pathogens, such as adenovirus, reovirus, astrovirus, infectious bursal disease and avian infectious bronchitis virus. These results showed that chicken anemia virus (CAV) may occur since the first few days of life in broilers - a fact not as yet reported -, associated with high pathogenic Infectious Bursal Disease Virus (IBDV) vaccine strain may induce a persistent growth retarded for several weeks in broilers.

Este estudo investigou a manifestação do vírus da Anemia Infecciosa das Aves (VAIA) em lotes de frangos que apresentavam retardo no crescimento e aumento da mortalidade observado a partir do quarto dia de idade. Clinicamente, as aves apresentavam depresão, palidez, despigmentação e retardo de crescimento. À necropsia, as aves apresentavam lesões compatíveis com a infecção pelo vírus da Anemia infecciosa das aves (VAIA). Amostras de fígado, baço e timo foram examinadas por PCR que amplifica um frangmento de 675 pb do gene VP-1 do VAIA. Todos os órgãos examinados foram positivos para o vírus da Anemia Infecciosa das Aves. Os demais patógenos, como adenovírus, reovírus, astrovírus, vírus da doença infecciosa bursal e coronavírus aviário não foram detectados pelas diferentes técnicas laboratoriais, como sorologia, PCR ou PAGE. Os resultados mostraram que o vírus da Anemia Infecciosa das Aves (VAIA) pode manifestar-se clinicamente nos primeiros dias de vida dos frangos – um fato ainda não reportado – associado ao vírus vacinal da doença infecciosa bursal (DIB) cepa forte pode induzir um persistente retardo de crescimento, por várias semanas, em frangos.

Porcine induced pluripotent stem cells produce chimeric offspring.

Ethical and moral issues rule out the use of human induced pluripotent stem cells (iPSCs) in chimera studies that would determine the full extent of their reprogrammed state, instead relying on less rigorous assays such as teratoma formation and differentiated cell types. To date, only mouse iPSC lines are known to be truly pluripotent. However, initial mouse iPSC lines failed to form chimeric offspring, but did generate teratomas and differentiated embryoid bodies, and thus these specific iPSC lines were not completely reprogrammed or truly pluripotent. Therefore, there is a need to address whether the reprogramming factors and process used eventually to generate chimeric mice are universal and sufficient to generate reprogrammed iPSC that contribute to chimeric offspring in additional species. Here we show that porcine mesenchymal stem cells transduced with 6 human reprogramming factors (POU5F1, SOX2, NANOG, KLF4, LIN28, and C-MYC) injected into preimplantation-stage embryos contributed to multiple tissue types spanning all 3 germ layers in 8 of 10 fetuses. The chimerism rate was high, 85.3% or 29 of 34 live offspring were chimeras based on skin and tail biopsies harvested from 2- to 5-day-old pigs. The creation of pluripotent porcine iPSCs capable of generating chimeric offspring introduces numerous opportunities to study the facets significantly affecting cell therapies, genetic engineering, and other aspects of stem cell and developmental biology.

Imaging diagnosis--split cord malformation.

The features of a calf with a split cord malformation are described. Clinically, there was severe cervicothoracic kyphoscoliosis and an interscapular dermal sinus associated with cerebrospinal fluid drainage. Using magnetic resonance imaging, complete duplication of the spinal cord at the cervical intumescence was detected. There was associated syringohydromyelia, multiple cervicothoracic vertebral malformations resulting in kyphoscoliosis and rachischisis, herniation of the cerebellar vermis, meningoencephalocele, and calvarial defects.

Neonatal chimerization with human glial progenitor cells can both remyelinate and rescue the otherwise lethally hypomyelinated shiverer mouse.

Congenitally hypomyelinated shiverer mice fail to generate compact myelin and die by 18-21 weeks of age. Using multifocal anterior and posterior fossa delivery of sorted fetal human glial progenitor cells into neonatal shiverer x rag2(-/-) mice, we achieved whole neuraxis myelination of the engrafted hosts, which in a significant fraction of cases rescued this otherwise lethal phenotype. The transplanted mice exhibited greatly prolonged survival with progressive resolution of their neurological deficits. Substantial myelination in multiple regions was accompanied by the acquisition of normal nodes of Ranvier and transcallosal conduction velocities, ultrastructurally normal and complete myelination of most axons, and a restoration of a substantially normal neurological phenotype. Notably, the resultant mice were cerebral chimeras, with murine gray matter but a predominantly human white matter glial composition. These data demonstrate that the neonatal transplantation of human glial progenitor cells can effectively treat disorders of congenital and perinatal hypomyelination.

Inactivation of the mouse Magel2 gene results in growth abnormalities similar to Prader-Willi syndrome.

Prader-Willi syndrome (PWS) is an imprinted genetic obesity disorder characterized by abnormalities of growth and metabolism. Multiple mouse models with deficiency of one or more PWS candidate genes have partially correlated individual genes with aspects of the PWS phenotype, although the genetic origin of defects in growth and metabolism has not been elucidated. Gene-targeted mutation of the PWS candidate gene Magel2 in mice causes altered circadian rhythm output and reduced motor activity. We now report that Magel2-null mice exhibit neonatal growth retardation, excessive weight gain after weaning, and increased adiposity with altered metabolism in adulthood, recapitulating fundamental aspects of the PWS phenotype. Magel2-null mice provide an important opportunity to examine the physiological basis for PWS neonatal failure to thrive and post-weaning weight gain and for the relationships among circadian rhythm, feeding behavior, and metabolism.

Perinatal ablation of the mouse lens causes multiple anterior chamber defects.

PURPOSE: The purpose of this study was to reassess the role of the lens as an "embryonic organizer" of ocular tissues. METHODS: We ablated the lens in mice by lens-specific expression of an attenuated version of diphtheria toxin A subunit(Tox176) driven by a modified crystallin promoter. Alterations in the differentiation programs of ocular tissues were examined by hematoxylin and eosin staining, in situ hybridization, and immunohistochemistry. RESULTS: Transgenic mice in the family OVE1757 exhibited severe microphakia. Apoptotic lens fibers were seen by embryonic day 15 (E15) and the lenses were completely ablated by post natal day 8. Multiple defects were seen in the anterior chamber. Corneal endothelial cells did not differentiate properly. The mesenchymal cells that would normally give rise to the endothelial layer were found to express N-cadherin, but they failed to form tight junctions and undergo a mesenchymal-to-epithelial transition. Although early specification of the presumptive ciliary body and iris was detected, subsequent differentiation of the iris was blocked. No dramatic changes were seen in the development of the retina. CONCLUSIONS: These results support the hypothesis that an intact lens is essential for proper differentiation of both the corneal endothelium and the iris and that the lens "organizes" the development of tissues in the anterior chamber.

Delineation of matriptase protein expression by enzymatic gene trapping suggests diverging roles in barrier function, hair formation, and squamous cell carcinogenesis.

The membrane serine protease matriptase is required for epidermal barrier function, hair formation, and thymocyte development in mice, and dysregulated matriptase expression causes epidermal squamous cell carcinoma. To elucidate the specific functions of matriptase in normal and aberrant epidermal differentiation, we used enzymatic gene trapping combined with immunohistochemical, ultrastructural, and barrier function assays to delineate the spatio-temporal expression and function of matriptase in mouse keratinized tissue development, homeostasis, and malignant transformation. In the interfollicular epidermis, matriptase expression was restricted to postmitotic transitional layer keratinocytes undergoing terminal differentiation. Matriptase was also expressed in keratinizing oral epithelium, where it was required for oral barrier function, and in thymic epithelium. In all three tissues, matriptase colocalized with profilaggrin. In staged embryos, the onset of epidermal matriptase expression coincided with that of profilaggrin expression and acquisition of the epidermal barrier. In marked contrast to stratifying keritinized epithelium, matripase expression commenced already in undifferentiated and rapidly proliferating profilaggrin-negative matrix cells and displayed hair growth cycle-dependent expression. Exposure of the epidermis to carcinogens led to the gradual appearance of matriptase in a keratin-5-positive proliferative cell compartment during malignant progression. Combined with previous studies, these data suggest that matriptase has diverging functions in the genesis of stratified keratinized epithelium, hair follicles, and squamous cell carcinoma.

Effect of hemi-circumferential periosteal transection and elevation in foals with experimentally induced angular limb deformities.

OBJECTIVE: To evaluate the effect of hemi-circumferential periosteal transection and elevation (HCPTE) in foals with, experimentally induced angular limb deformities. DESIGN: Prospective study. ANIMALS: 10 healthy foals. PROCEDURE: When foals were 30 days old, transphyseal bridge implants were placed on the lateral aspects of both distal radial physes. At 90 days of age (or when 15 degrees of angulation had developed), implants were removed, and HCPTE was performed on 1 limb. Foals were confined in small pens after surgery; the front feet of the foals were rasped weekly to maintain medial-to-lateral hoof wall balance. Dorsopalmar radiographic projections of the carpi were obtained before HCPTE and 2, 4, 6, 8, and 48 weeks later. RESULTS: At the time of transphyseal bridge removal and HCPTE, both treated and control limbs were observed to have a significantly greater carpal valgus, compared with the initial degree of angulation at 30 days of age. Following HCPTE or sham surgery, all limbs straightened over the subsequent 2 months of the study. Median angulation was not significantly different between treated and control limbs at any time during the study. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that in foals with experimentally induced limb deformities, HCPTE was no more effective than stall confinement and hoof trimming alone for correction of the deformity.

Matriptase/MT-SP1 is required for postnatal survival, epidermal barrier function, hair follicle development, and thymic homeostasis.

Matriptase/MT-SP1 is a novel tumor-associated type II transmembrane serine protease that is highly expressed in the epidermis, thymic stroma, and other epithelia. A null mutation was introduced into the Matriptase/MT-SP1 gene of mice to determine the role of Matriptase/MT-SP1 in epidermal development and neoplasia. Matriptase/MT-SP1-deficient mice developed to term but uniformly died within 48 h of birth. All epidermal surfaces of newborn mice were grossly abnormal with a dry, red, shiny, and wrinkled appearance. Matriptase/MT-SP1-deficiency caused striking malformations of the stratum corneum, characterized by dysmorphic and pleomorphic corneocytes and the absence of vesicular bodies in transitional layer cells. This aberrant skin development seriously compromised both inward and outward epidermal barrier function, leading to the rapid and fatal dehydration of Matriptase/MT-SP1-deficient pups. Loss of Matriptase/MT-SP1 also seriously affected hair follicle development resulting in generalized follicular hypoplasia, absence of erupted vibrissae, lack of vibrissal hair canal formation, ingrown vibrissae, and wholesale abortion of vibrissal follicles. Furthermore, Matriptase/MT-SP1-deficiency resulted in dramatically increased thymocyte apoptosis, and depletion of thymocytes. This study demonstrates that Matriptase/MT-SP1 has pleiotropic functions in the development of the epidermis, hair follicles, and cellular immune system.

Umbilical evagination of the urinary bladder in a neonatal filly.

An 8-hour-old Standardbred filly was evaluated because of an enlarging umbilical mass and stranguria. It was suspected that the mass was the urinary bladder; this was confirmed on surgical exploration of the abdomen. Despite a normal umbilical ring, the bladder had descended and partially everted through its urachal communication with the umbilical stalk. Partial cystectomy and umbilical resection were performed and resulted in an excellent clinical outcome. Evagination of the urinary bladder via the umbilicus has rarely been described in human infants, and, to our knowledge, it has not been reported in the veterinary literature.

Abnormal iron accumulation in the brain of neonatal hypotransferrinemic mice.

Transferrin is a plasma protein involved in iron delivery to tissues. To study iron transport into the brain under a transferrin deficiency, iron concentration and 59Fe uptake in the brain were measured in neonatal hypotransferrinemic (HP) mice at 7 days of age. Brain iron concentration of the HP mice, in which iron concentration was relatively high in the cerebral cortex and cerebellum, was approximately three times higher than that of non-mutant mice, whereas serum iron concentration of HP mice was significantly lower than that of non-mutant mice. When 59FeCl3 was subcutaneously injected into HP and non-mutant mice, 59Fe was distributed highly in the choroid plexus in the ventricles of HP mice 24 h after injection. The 59Fe distribution in the brain was different between HP and non-mutant mice. On the other hand, the clearance of 59Fe from the blood was very high in HP mice and the hepatic 59Fe concentration of HP mice was more than ten times of that of non-mutant mice. The present findings demonstrate that iron distribution in the brain is changed by transferrin deficiency and that iron abnormally accumulates in the brain of HP mice. It is likely that the management of iron is different in the brain of HP mice.

Anophthalmia in litters of female rats treated with the food-derived carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine.

Anophthalmia in litters of pregnant rats treated with 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a food-derived carcinogen, was incidentally encountered in a risk-assessment study with 2-generation exposure to PhIP. Female Fischer 344 animals were given 200 ppm PhIP in the diet for 4 wk before mating with nontreated males and also during gestation and lactation periods. Mean numbers of newborn rats per litter in control and PhIP-treated groups were 7.9 +/- 2.9 and 7.1 +/- 1.6 in trial 1 and 8.3 +/- 1.9 and 6.1 +/- 2.4 in trial 2. Among 49 (trial 1) and 63 (trial 2) offspring from PhIP-treated dams, 9 (18.4%) and 32 (50.8%) demonstrated anophthalmia, and 1 (2.0%) and 8 (12.7%) demonstrated hydrocephaly. Five of 7 (71.4%) and 13 of 14 (92.9%) dams delivered pups with malformations in trials 1 and 2, respectively. Also, in a previous study that was carried out with the same protocol and that used the Sprague-Dawley strain of rats, anophthalmia and hydrocephaly were observed in 2 and 1 out of 175 pups, respectively, from 100 ppm PhIP-treated dams. No congenital malformations were found in control groups of the same size in either experiment. In addition to having been previously identified as a cause of carcinogenic activity, our findings suggest that PhIP is capable of causing anophthalmia in rats when administered during the gestational period.

Management of bilateral flexural deformity of the metacarpophalangeal joints in three alpaca crias.

Three neonatal alpacas were treated for MCP flexural deformities. Two crias responded well to conservative therapy using splints made from fibreglass cast material. One cria with severe deformity failed to respond to conservative treatment but recovered satisfactorily after transection of the suspensory ligament in both legs. It seems that mild cases of flexural deformity of the MCP joint in alpaca crias respond well to conservative therapy but that surgical correction may be required in more severe cases. Sequential transection of the structures limiting extension of the MCP joint may result in fewer complications than previously reported techniques.

Perioperative management and mortality rates of dogs undergoing cesarean section in the United States and Canada.

OBJECTIVE: To describe dogs undergoing cesarean section in the United States and Canada, to determine perioperative management, and to calculate survival proportions. DESIGN: Multicenter prospective case series. ANIMALS: 3,908 puppies from 808 dams. RESULTS: Survival rates immediately, 2 hours, and 7 days after delivery were 92, 87, and 80%, respectively, for puppies delivered by cesarean section (n = 3,410) and 86, 83, and 75%, respectively, for puppies born naturally (498). For 614 of 807 (76%) litters, all puppies delivered by cesarean section were born alive. Maternal mortality rate was 1% (n = 9). Of 776 surgeries, 453 (58%) were done on an emergency basis. The most common breeds of dogs that underwent emergency surgery were Bulldog, Labrador Retriever, Boxer, Corgis, and Chihuahua. The most common breeds of dogs that underwent elective surgery were Bulldog, Labrador Retriever, Mastiff, Golden Retriever, and Yorkshire Terrier. The most common methods of inducing and maintaining anesthesia were administration of isoflurane for induction and maintenance (n = 266; 34%) and administration of propofol for induction followed by administration of isoflurane for maintenance (237; 30%). CLINICAL IMPLICATIONS: Mortality rates of dams and puppies undergoing cesarean section in the United States and Canada are low. Knowledge of mortality rates should be useful to veterinarians when advising clients on the likelihood of puppy and dam survival associated with cesarean section.

Congenital defects in newborn foals of mares treated for equine protozoal myeloencephalitis during pregnancy.

Three weak, recumbent neonatal foals with skin lesions, including a thin wooly coat, were born to mares being treated for equine protozoal myeloencephalitis. Mares received sulfadiazine or sulfamethoxazole-trimethoprim, pyrimethamine, folic acid, and vitamin E orally. Foals were anemic, leukopenic, azotemic, hyponatremic, and hyperkalemic. Serum folate concentrations in the 3 foals and 2 mares were lower than those reported in the literature for clinically normal brood mares. Treatment was unsuccessful. For each foal, necropsy revealed lobulated kidneys with thin cortices and a pale medulla, and the spleen and thymus were small. Histologic examination revealed marked epidermal necrosis without inflammatory cells, thin renal cortices, renal tubular nephrosis, lymphoid aplasia, and bone marrow aplasia and hypoplasia. These observations indicate that oral administration of sulfonamides, 2,4-diaminopyrimidines (pyrimethamine with or without trimethoprim), and folic acid to mares during pregnancy is related to congenital defects in newborn foals.

Homeosis and intestinal tumours in Cdx2 mutant mice.

In Drosophila, disturbing the expression of the homeobox gene caudal causes a severe disruption in body segmentation and global body patterning. There are three mouse homologues of Drosophila caudal: Cdx1 (ref. 2), Cdx2 (ref. 3) and Cdx4 (ref. 4). We have generated a null mutation of murine Cdx2 by homologous recombination. Cdx2 homozygote null mutants die between 3.5 and 5.5 days post coitum (d.p.c.). Cdx2 heterozygote mutants exhibit a variable phenotype, with many showing tail abnormalities or stunted growth. Skeletal analysis demonstrates a homeotic shift of vertebrae and compatible malformations of the ribs. Within the first three months of life, 90% of Cdx2 heterozygotes develop multiple intestinal adenomatous polyps, particularly in the proximal colon. These polyps occasionally contain areas of true metaplasia. In contrast to the surrounding intestinal epithelium, the neoplastic cells do not express Cdx2 from the remaining allele. These results suggest that Cdx2 mutation is the primary event in the genesis of some intestinal tumours.