Breed differentiation among Japanese native chickens by specific skull features determined by direct measurements and computer vision techniques.

1. Inter-breed morphological comparisons were made among 11 breeds of Japanese native chickens (Gifujidori, Hinaidori, Shokoku, Totenko, Tomaru, Satsumadori, Shamo, Koshamo, Koeyoshi, Chabo and Nagoya), White Leghorn, broiler chickens (Chunky) and red junglefowl collected in the Philippines, based on results of direct measurements and analysis by computer vision techniques of the skull. 2. Analysis of direct measurements identified two groups of chicken: a small type that included the Chabo, Koshamo, red junglefowl, Gifujidori and Shokoku and a large type that included the remaining breeds studied. These groupings were made based on size determined both in the first (PC1) and second principal component (PC2). The greatest length of the cranium and condylobasal length greatly contributed to the morphological differences between these two groups. 3. Analysis by computer vision techniques, however, identified three groups: the Bantam group (which includes red junglefowl), Shokoku group and Shamo group. White Leghorn clustered within the Shokoku group while the broiler chicken belonged to the Shamo group. The region around the junction of the neural cranium and the visceral cranium contributed greatly to the morphological differences among breeds, both in the PC1 and PC2.

A light and electron microscopical study of the Tragulid (mouse deer) placenta.

The Tragulidae are the living relics of the basal ruminant stock. They have a diffuse placenta, with no aggregations of the placental villi into localised placentomes characteristic of all other ruminants. Despite this difference, this ultrastructural and immunocytochemical investigation demonstrates that in Tragulus the trophoblast binucleate cell (BNC) plays the same central role in development and structure as in all other ruminants. It shows an identical development and ultrastructure, produces granules reactive with bovine placental lactogen and pregnancy associated glycoprotein antibodies, and migrates when mature through the trophoblast tight junction to fuse into a mosaic of syncytial plaques from which the granules are released to the mother and which have replaced the uterine epithelium. Unlike the persistent plaques in the sheep and goat placenta, in Tragulus they are transient, dying by apoptosis with the fragments phagocytosed by the trophoblast. This brings the trophoblast into direct endotheliochorial apposition to maternal tissue until BNC migration and fusion replace the dead plaque. This intimate fetomaternal confrontation has not been shown in any other ruminant, and could be a relic of the evolutionary development of the synepitheliochorial from the original basic eutherian endo- or hemo-chorial placenta.

Exendin-4, glucagon-like peptide-1 receptor agonist, enhances isoflurane-induced preconditioning against myocardial infarction via caveolin-3 expression.

OBJECTIVE: To investigate the cardioprotective effects of isoflurane and exendin-4 against myocardial ischemia/reperfusion injury and the signaling pathways through which these effects are mediated. MATERIALS AND METHODS: For infarct size measurements, anesthetized mice were subjected to 30 min of coronary artery occlusion followed by 2 h of reperfusion. Wild-type or caveolin-3 knockout mice received isoflurane, exendin-4, or isoflurane with exendin-4 before ischemia index determination. Caveolin-3 expression in the heart was measured by immunoblotting. RESULTS: Myocardial infarct size was smaller in the isoflurane- [1.0 minimum alveolar concentration (MAC)] or exendin-4- (30 ng/kg i.v.) treated groups than the controls. Infarct size was not affected by isoflurane at 0.5 MAC or 3 ng/kg i.v. exendin-4, but the combination of these treatments reduced infarct size. Pharmacological preconditioning (isoflurane at 1.0 MAC, 30 ng/kg i.v. exendin-4, or isoflurane at 0.5 MAC with 3 ng/kg i.v. exendin-4) increased caveolin-3 protein expression in the heart after infarct induction. The cardioprotective effects of isoflurane, exendin-4, and isoflurane with exendin-4 were abolished in caveolin-3 knockout mice. CONCLUSIONS: The combination of isoflurane and exendin-4 reduced infarct size, but it was not more effective than either agent alone, and the cardioprotective effects of these agents are mediated by caveolin-3 expression.

Anatomical and histological characterization of the female reproductive organs of mouse deer (Tragulidae).

The Tragulidae may be a type that represents the earliest ruminant families to evolve. The female reproductive organs including ovary, oviduct, uterus and placenta were anatomically and histologically investigated. The structures of the ovary and oviduct were very similar to other ruminants. However, the gross structure of the placenta was diffuse and thus noticeably different from other ruminants which are polycotyledonary. Histologically, the placenta of Tragulidae appears to be epitheliochorial and therefore similar to other ruminants. Numerous trophoblastic binucleated cells which are characteristic of all other ruminants were observed. These results suggest that the placenta of Tragulidae is a transitional type between diffuse epitheliochorial and polycotyledonary synepitheliochorial categories.

Reconstruction of the anterior cranial base with the galeal frontalis myofascial flap and the vascularized outer table calvarial bone graft.

Reconstruction of the anterior cranial base after tumor extirpation must seal off the cranial cavity from the upper respiratory tract. The key to success is to use vascularized materials for the structural support of the brain. From October 1989 to July 1992, 10 patients underwent anterior cranial base reconstruction after basicranial tumor resection; the lesions were four meningiomas and six malignant tumors of the ethmoid, maxilla, and orbit. The malignant tumors included four recurrent tumors that had been previously treated by a transfacial approach. After tumor extirpation, the resultant bony defects in the anterior cranial base, involving the orbital roof as well as the cribriform plate, ranged from 4 x 3 to 6 x 7 cm in size. The materials used in reconstruction were the galeal frontalis myofascial flap and the outer table calvarial bone flap, which is based on the temporoparietal galeal flap. Both materials are known to have rich blood supplies. These flaps make a reliable separation between the cranial cavity and the respiratory tract in three layers: the galeal frontalis myofascial flap, the vascularized calvarial bone, and the temporoparietal galea. Postoperative complications included one subcutaneous hematoma and one temporary cerebrospinal fluid rhinorrhea. We think this reconstructive technique will be useful in selected circumstances, especially after resection of a recurrent malignant tumor.

Benign fibroosseous lesions involving the skull base, paranasal sinuses, and nasal cavity. Report of two cases.

The authors report two patients with benign fibroosseous lesions involving the center of the skull base: a 15-year-old boy with repetitive meningitis and pneumocephalus and a 11-year-old boy with nasal obstruction and headache. The clinical diagnoses were ossifying fibroma and aneurysmal bone cyst, respectively. Lesions in both patients extended to the nasal cavity, the sphenoid and posterior ethmoid sinuses, and the skull base, where the planum sphenoidale, the sella turcica, the upper two-thirds of the clivus, and the medial portion of the middle cranial base were involved. The lesions were totally removed using an anterior craniofacial approach. Characteristics of these lesions and the surgical approach are discussed.

Isolation of a germline-transmissible embryonic stem (ES) cell line from C3H/He mice.

We have isolated three embryonic stem (ES) cell lines from C3H/He mice using mouse STO cells as a feeder layer. One ES cell line (H-1) was male, and two (H-2 and H-3) were female, as determined by polymerase chain reaction, in situ hybridization, and karyotype analyses. All were immunocytochemically reactive with a C3H strain-specific antibody. Injection of cells from the female ES H-3 line into C57BL/6 blastocysts yielded four chimeras with slight coat color chimerism. All chimeras were male, and as expected, no germline-transmission was observed. By contrast, when male ES H-1 cells were injected into the perivitelline space of 8-cell C57BL/6 embryos, one male mouse with overt coat color chimerism was recovered, and it produced ES H-1-derived offspring exclusively. This germline-transmissible C3H/He cell line represents a novel addition to those ES lines currently employed for gene manipulation studies of development.

Purification of rabbit blood basophils by isopycnic banding on Percoll.

This study defines the basophil densities in rabbits and also describes a relatively simple method of purifying basophils from peripheral blood through a two-step procedure. To evaluate the recovery of basophils, EDTA-anticoagulated blood in layered over Percoll of various densities, ranging from 1.070 to 1.080 g/ml. The cells retained in Percoll were collected separately after centrifugation at 600 g(av) for 30 min (Step I). Distribution of basophils indicated that basophils had densities of between 1.070 and 1.078 g/ml, with a peak at 1.074 g/ml. To enhance the basophil purity, a second centrifugation of the cells obtained from Step I over appropriate Percoll gradient led to further enhancement of basophil purity (30.4 +/- 0.7%) which was (20.6 +/- 0.2%) in Step I. The cells obtained by this method appeared morphologically normal and viable.

Surgical treatment of a massive chondrosarcoma in the skull base associated with Maffucci's syndrome: a case report.

BACKGROUND: A successfully treated massive chondrosarcoma in the skull base associated with Maffucci's syndrome is presented. The purpose of this report is to discuss the surgical approach to the tumor and reconstruction of the skull base. CASE DESCRIPTION: A 36-year-old woman who had a history of multiple enchondromas and subcutaneous hemangiomas presented with decreased right visual acuity and left papilledema. Computed tomography (CT) and magnetic resonance imaging (MRI) demonstrated a mass in the skull base. The tumor occupied the nasal and paranasal cavities, and extended to the anterior, middle, and posterior intracranial spaces. The midline skull base structures and the left middle cranial base were destroyed. Using a combined anterior craniofacial and left orbitozygomatic approach, the tumor was totally resected. The large skull base defect was reconstructed with a vascularized outer table parietal bone graft attached to a bipedicled temporoparietal galeal flap. The postoperative course was uneventful except for decreased left visual acuity, and temporary diplopia and facial hypesthesia. In 40 months of follow-up there was no recurrence. CONCLUSIONSA skull base approach should be selected to perform total resection of an extensive skull base tumor. The bipedicled temporoparietal galeal flap and vascularized calvarial bone was useful for simultaneous reconstruction.

Surgical approach to midline skull base tumors with olfactory preservation.

Through the frontal approach, a skull base tumor in the deep midline region is obscured by the cribriform plate. This paper demonstrates our experience of operating on seven patients with midline skull base tumors posterior to the cribriform plate using a technique of maintaining the olfactory-cribriform anatomy. Following a bifrontal craniotomy and an orbitonaso-glabellar osteotomy, a circumferential osteotomy of the cribriform plate and horizontal incision of the nasal mucosa permitted mobilization of the cribriform plate unit along with the frontal lobe, providing a wide exposure of the posterior midline area. Tumors were resected from the posterior nasopharynx, ethmoid sinus, sphenoid sinus, and upper portion of the clivus. The skull base defects were repaired with the bipedicled temporoparietal galeal flap. Two patients lost olfaction because of poor preservation of the nasal mucosa and intradural dissection of the olfactory bulb. Five patients showed complete recovery of olfaction in 8 weeks. None of the patients developed cerebrospinal fluid leakage, infection, or increased neurologic deficits. En bloc mobilization of the cribriform plate and the bipedicled temporoparietal galeal flap are very useful for the management of midline cranial base tumors posterior to the cribriform plate as long as the olfactory unit is not involved with tumors.

Arteriovenous fistula formation after punch graft hair transplantation in the frontal region.

A case of arteriovenous fistula formation following punch hair grafting is presented. This is indeed a potential complication of a hair plug transplantation procedure in both recipient and donor sites. Surgical excision of the fistula in the frontal or midline scalp region requires resection of the overlying skin because the involved vessels lie within the subdermal plexus.

Efficacy of cycled hyperinflation for rapid tissue expansion.

Temporary hyperexpansion at each filling session has previously been confirmed to increase the inflation volume in a conventional expansion procedure. This study employed eight young pigs to evaluate the effect of hyperinflation of this type in rapid expansion performed on 10 consecutive days. Hyperinflation to 120 mmHg for 4 minutes and deflation to 40 mmHg for 2 minutes was repeated five times on the experimental side. This cycled hyperinflation was found to increase the filling volume by 49 percent when compared with the contralateral side, which was not cycled or hyperinflated (control) (p < 0.01). The increase in surface area of the entire skin over the balloon was 178.1 percent in the hyperinflation group and 132.8 percent in the controls (p < 0.01). The actual expansion in surface area was not significantly different: 44.4 percent in the hyperinflation group and 45.7 percent in the control group. The additional gain of skin by cycled hyperinflation was the result of recruitment. The control group required 5 more days to achieve the same amount of skin but resulted in more true expansion and less recruitment. It is concluded that the cycled hyperinflation technique may shorten the time period of the whole reconstruction procedure but may produce an aesthetic deformity and flap necrosis.

A cadaver investigation of the blood supply of the galeal frontalis flap.

The vascular anatomy of the galeal frontalis flap was studied in 12 fresh cadavers by an intraarterial dye injection technique. Special attention was directed to the length limit of this flap. The general belief that the galeal frontalis flap has a robust vascularity by means of the supratrochlear and supraorbital arteries was not demonstrated in this study. In the medial half of the forehead, superficial branches of both arteries penetrated the frontalis muscle immediately above the supraorbital rim and ran superficially in the subcutaneous tissue. In the lateral half, some of the superficial branches of the supraorbital artery traveled with the frontalis muscle and anastomosed with the frontal branch of the superficial temporal artery. Deep branches of the supratrochlear and supraorbital arteries showed an axial distribution on the periosteum only for a short distance. One or two branches of the supraorbital artery were found to take a superficial course within the subgaleal layer, pierce the frontalis muscle, and anastomose with the superficial temporal artery. These findings suggest that the galeal frontalis flap should be elevated in the lateral forehead. The preservation of the periosteum with the flap is recommended in order to ensure the temporoparietal extension.

The vascular anatomy of the galeal flap in the interparietal and midline regions.

The potential extension of the galeal flap in the interparietal area was studied on 17 fresh human cadaver heads by intravascular dye injection technique. It was demonstrated that an ipsilateral superficial temporal artery that supplies the galeal flap does not cross the midline or anastomose with the contralateral superficial temporal artery but ensures the survival of a flap extended up to 1 cm proximal to the sagittal suture line. The width of the temporoparietal flap can be extended up to 15 cm, depending on the vascular pattern of the superficial temporal artery. When required, the lateral extension may provide the required soft-tissue bulk despite the reduced flap length.

Distribution of neuropeptide-immunoreactive nerve fibres in the medulla oblongata of the chicken.

The anatomical distribution of nerve fibres containing galanin (GAL), leucine enkephalin (lENK), methionine enkephalin (mENK) and substance P (SP) was investigated in the medulla oblongata of the chicken by means of immunohistochemistry. In the medulla oblongata, dense networks of mENK-, lENK-, SP- and GAL-immunoreactive nerve fibres were mainly restricted within and around the dorsal vagal complex, i.e., the nucleus of the solitary tract and dorsal motor nucleus of the vagus nerve. The relative density of immunoreactive nerve fibres was usually decreased in the order of mENK-, SP-, lENK- and GAL-immunoreactive ones. Immunoreactive nerve fibres for each antiserum had a unique distribution among the subnuclear groups of the dorsal vagal complex. Sparser networks of immunolabelled nerve fibres were found in the nucleus of the hypoglossal nerve, nucleus ambiguus, nucleus of the descending trigeminal tract, raphe nucleus and caudal olivary complex. In the colchicine-pretreated chicken, some nerve cell bodies expressing the immunoreaction for ENK and GAL antisera were detected in the nucleus of the solitary tract and nucleus of the descending trigeminal tract. There were some differences in the distribution pattern of immunoreactive nerve elements among the four kinds of antisera. The selective distribution of neuropeptide-containing nerve elements within various nuclei of the medulla oblongata suggests that these neuropeptides may be differentially involved in central neural regulation.

Ultrastructure of enkephalin- and substance P-immunoreactive axon terminals in the hypoglossal nucleus of the chicken.

The morphology and distribution of leucine-enkephalin (LENK)- and substance P (SP)-containing axon terminals in the hypoglossal nucleus of the chicken was investigated immunocytochemically at the electron microscopic level. The major findings of the present study were as follows: 1) Both LENK- and SP-immunoreactive (IR) profiles including unmyelinated axons, varicosities and axon terminals were found throughout the hypoglossal nucleus. 2) LENK-IR profiles without synapses (preterminal axons and varicosities) were observed more frequently (66%) than LENK-IR synaptic profiles (34%). In contrast, SP-IR synaptic terminals were seen more often (68%) than SP-IR terminals without synapses (32%). 3) Both LENK- and SP-IR synaptic terminals ending on dendrites were more numerous (83% and 94%, respectively) than those making synapses on somata (17% and 6%, respectively). 4) The average diameter of SP-IR synaptic terminals were significantly larger than that of LENK-IR ones. 5) Both LENK- and SP-IR synaptic terminals contained numerous small clear vesicles and several large dense-cored vesicles (LDV). Although both LENK- and SP-IR synaptic terminals contained LDV consistently, SP-IR synaptic terminals contained significantly more LDV than LENK-IR types. 6) Postsynaptic dense bodies (Taxi bodies) were seen more frequently in SP-IR synaptic terminals than in LENK-IR ones. These findings confirm both LENK and SP innervation in the hypoglossal nucleus and suggest a functional role as neuromodulator for the two neuropeptides in tongue control.

Segmental mandibular reconstruction by distraction osteogenesis under skin flaps.

In five adult dogs, molars were extracted and skin flaps from the neck prepared for delayed transplantation. Two weeks later, a 25-mm segment of the mandible was excised with surrounding periosteum and gingiva. The mandible was stabilized with a reconstruction plate and the intraoral defect repaired with a pedicled skin flap. A proximal transport segment was created and an external distraction device was applied. After one week, distraction of the transport segment was begun at a rate of 1 mm/day. After distraction was completed, the lengthening appliance was left in place for 12 weeks until the dogs were killed. Radiologic and histologic examination revealed new bone at the distraction site. The intraoral skin flap was pushed out of the defect as distraction progressed. Bony union of the transport segment to the distal stump was not achieved due to intervening soft tissue. These results suggest that it is feasible to bridge a mandibular defect that is covered with a skin flap, with distraction osteogenesis.

Effect of insulin on in vitro development of tetraploid mouse embryos.

The effect of insulin on in vitro development of early stage tetraploid mouse embryos was examined. Tetraploid embryos were produced at the 2-cell stage by treatment with cytochalasin B for 12 hr, then they were cultured with M16 medium containing 1 microM insulin. Although insulin had no effect on the embryos until 72 hr after hCG injection, at 96 hr the number of cells of insulin (+) tetraploid blastocysts increased 20.2% more than insulin (-) tetraploid embryos. The increase in the number of cells mainly depended on the inner cell mass (ICM), in which insulin (+) cells increase 76.3% more than insulin (-) cells. These effects of insulin on tetraploid embryos were comparable with those on diploid embryos. The present results suggested that the effect of insulin on the cell proliferation of early stage embryos was not affected by tetraploidy, but the increased number of cells in tetraploid blastocysts was significantly smaller than that in diploid embryos.

Preimplantation development of tetraploid mouse embryo produced by cytochalasin B.

Tetraploid mouse embryos usually cease to develop early after implantation, though they can develop to blastocysts. To characterize the failure of development in detail, tetraploid mouse embryos at the preimplantation period were examined as to both their morphology and number of cells. The tetraploid embryos were produced by 12 hr treatment with cytochalasin B (CB) at the 2-cell stage of backcross of (C57BL/6 x C3H/He) F1 x C3H/He. The tetraploid embryos in the preimplantation period exhibited compaction at 72 hr after hCG injection and blastocyst formation at 96 hr, as well as diploid embryos, but the number of cells composing the embryos was significantly smaller than that in the diploid embryos. At the term 60-96 hr after hCG injection, mean cell cycles were 14.03 hr in the tetraploid embryos, but 12.02 hr in the diploid. When tetraploid embryos were transferred into the oviducts of pseudopregnant recipients immediately after CB treatment, the number of cells in tetraploid blastocysts was increased compared with the embryos cultured in vitro, though the number did not reach that of diploid embryos. These results suggested that compaction and blastocyst formation in preimplantation development of tetraploid embryos depended on the time after hCG injection, irrespective of the number of cells or the length of the cell cycle. The lengthening of the cell cycle in tetraploid embryos may be one of the causes of failure in postimplantation development.

Retinal projections to the subcortical nuclei in the Japanese field vole (Microtus montebelli).

Retinal projections in the Japanese field vole (Microtus montebelli) were determined by anterograde transport of horseradish peroxidase (HRP). Injection of HRP into the unilateral vitreous body demonstrated that the terminal labeling of the optic projections was seen bilaterally in the suprachiasmatic nucleus (SCH), the ventral (GLv) and dorsal (GLd) lateral geniculate nuclei, the intergeniculate leaflet (IGL), the medial pretectal nucleus (NTOM) of the pretectum (PT) and the superficial layer of the superior colliculus (CS), with contralateral predominance, and only contralaterally labeled terminals were found in the lateroposterior thalamic nucleus (LP), the lateral pretectal nucleus (NTOL) of the PT, the dorsal (DTN) and medial (MTN) terminal nuclei of the accessory optic system (AOS). The distribution area of the retinofugal terminals was divided into a three laminar arrangement in the GLd, i.e., layers 1 and 3 and layer 2, received the retinal input from contralateral and ipsilateral eye, respectively, as in arboreal squirrels. The contralateral CS received retinal fibers in the superficial layer, while ipsilateral optic fibers projected sparsely to the stratum opticum of the colliculi. Retinal connections to the DTN and MTN of the AOS were clearly discerned but no lateral terminal nucleus with retinal afferents was found. In addition, the AOS had no inferior fasciculus. These findings indicate that the vole has a contradictory features of a well- and a less-developed sense of vision. Namely, the image forming visual system such as the retino-GLd was as well-developed as in a squirrel, on the other hand, the non-image forming visual system such as the retino-AOS was less-developed as in an insectivore's brain.