Genetic Diversity of mtDNA D-loop Polymorphisms in Laotian Native Fowl Populations.

Here, we studied the genetic diversity of native fowls in Laos by analyzing a mitochondrial DNA (mtDNA) sequence polymorphism. A 546-bp fragment of the mtDNA D-loop region was sequenced in 129 chickens from the areas of Vientiane, Luang Prabang and Pakse. In total, 29 haplotypes were identified and formed five clades. Haplotype diversity and nucleotide diversity of the native fowls in Laos were 0.85536±0.0172 and 0.010158±0.005555, respectively. Although the Laotian native fowls were distributed across five clades, most of them were clustered in two main clades (A and B), which were originated in China. The other haplotypes were contained in clades D, F, and I, which originated from continental southeast Asia. These results suggest that multiple maternal lineages were involved in the origin of domestic chicken in Laos. Moreover, there appear to be at least two maternal lineages, one from China and the other from the southeast Asian continent.

Relationship between abdominal fat content and avian uncoupling protein gene expression in skeletal muscle of Japanese quail Coturnix japonica.

1. The genetic architecture of the avian uncoupling protein (avUCP) was investigated and the relationship between avUCP gene expression and the amount of abdominal fat of Japanese quail was determined by quantitative real-time PCR. 2. The Japanese quail avUCP gene consists of six exons and five introns. Sequences of nucleotides and amino acids were 94·6% and 86·0% identical to those of the chicken avUCP gene, and phylogenetic analysis showed that the Japanese quail avUCP gene consists of the same clusters as the chicken and turkey avUCP. 3. Expression of the avUCP gene was significantly higher in the Pectoralis major (1·28 ± 0·24) than in the Biceps femoris (0·63 ± 0·14). 4. A positive correlation coefficient between the avUCP gene expression in the Pectoralis major and Biceps femoris was observed (r = 0·79, P = 0·02), whereas a negative correlation coefficient was observed between the abdominal fat percentage (AFP) and gene expression in both the Pectoralis major (r = -0·82, P = 0·01) and Biceps femoris (r = -0·61, P = 0·11). 5. The avUCP gene was associated with the accumulation of abdominal fat in Japanese quail and it was concluded that modulation of avUCP gene expression could be utilised to control abdominal fat accumulation in poultry.

Primary malignant lymphoma of the maxillary sinus: CT and MRI.

We reviewed the CT and MRI of seven patients with primary malignant lymphoma of the maxillary sinus to find if there are characteristic imaging findings suggestive of the disease. The images were analysed for appearance, size, signal, internal characteristics, extent of tumour, bone change and lymph node enlargement. In two patients, the tumour first presented with mucosal thickening. In the remaining five, the tumours were an expansile mass 4-6 cm in diameter at the time of detection. Although it was difficult to distinguish tumour from mucosa or obstructed fluid on CT, T2-weighted MRI enabled us to separate tumour from normal mucosa or fluid. In two patients, the tumours were heterogeneous. Calcification and haemorrhage were observed in one patient. Periantral soft-tissue infiltration was always present, even when tumour appeared as slight mucosal thickening. Posterior extension was seen in all patients. Permeative and lytic bone destruction accompanied most cases of periantral soft-tissue infiltration; mixed destruction and sclerosis was also observed. Mucosal thickening with periantral soft-tissue infiltration may suggest malignant lymphoma of the maxillary sinus in its early form. Various types of bone change may accompany the periantral soft-tissue infiltration.

New cosmid vectors for library construction, chromosome walking and restriction mapping in filamentous fungi.

New cosmid vectors were constructed for the ascomycete fungus, Magnaporthe grisea and the basidiomycete fungus, Ustilago maydis. These vectors are capable of transforming M. grisea at frequencies of up to 5 transformants/micrograms linear DNA and U. maydis at up to 25 transformants/microgram circular DNA for integrative transformation. In addition, 2800 transformants/microgram DNA are possible when using an autonomously replicating vector. Since the promoters used in these vectors function in other ascomycete and basidiomycete fungi, we anticipate that these vectors will be widely applicable.

The Magnaporthe grisea DNA fingerprinting probe MGR586 contains the 3' end of an inverted repeat transposon.

The Magnaporthe grisea repeat (MGR) sequence MGR586 has been widely used for population studies of the rice blast fungus, and has enabled classification of the fungal population into hundreds of genetic lineages. While studying the distribution of MGR586 sequences in strains of M. grisea, we discovered that the plasmid probe pCB586 contains a significant amount of single-copy DNA. To define precisely the boundary of the repetitive DNA in pCB586, this plasmid and four cosmid clones containing MGR586 were sequenced. Only 740 bp of one end of the 2.6-bp insert in the pCB586 plasmid was common to all clones. DNA sequence analysis of cosmid DNA revealed that all the cosmids contained common sequences beyond the cloning site in pCB586, indicating that the repetitive DNA in the fingerprinting clone is part of a larger element. The entire repetitive element was sequenced and found to resemble an inverted repeat transposon. This putative transposon is 1.86 kb in length and has perfect terminal repeats of 42 bp, which themselves contain direct repeats of 16 bp. The internal region of the transposon possesses one open reading frame which shows similarity at the peptide level to the Pot2 transposon from M. grisea and Fot1 from Fusarium oxysporum. Hybridization studies using the entire element as a probe revealed that some strains of M. grisea, whose DNA hybridized to the pCB586 probe, entirely lacked MGR586 transposon sequences.

Aortic lesions in nonlaying hens with endogenous hyperlipidemia.

Atherogenicity of chemically discrete cholesterol has been questioned earlier at small doses insufficient to cause hypercholesterolemia. Long-term experiments involving large doses are hindered by the propensity of cholesterol to autoxidize in air. As an alternative, aortic lesions in mature, 1- to 3-year-old laying and hereditary nonlaying hens were studied by electron microscopy. Due to spontaneous egg yolk involution, serum cholesterol values of nonlayers were high, averaging 583 mg/dL, and triglyceride values averaged 1,777 mg/dL. Respective values in layers were 106 mg/dL and 894 mg/dL. Aortic lesions in nonlayers developed earlier and were more extensive and fatty than in layers. Lipids were stainable even in the media of grossly normal segments of aorta in nonlayers. Findings suggested an important role of inhibition of hyperlipidemic components. It is unknown, however, whether large amounts of endogenous cholesterol in serum undergo autoxidation in vivo.

Vitamin D-induced coronary atherosclerosis in normolipemic swine: comparison with human disease.

Coronary atherosclerosis developed in normolipemic swine fed a basal ration supplemented with 125,000 IU, 62,500 IU and 31,250 IU of vitamin D3/kg of diet for 3 months and subsequently only the basal ration for the following 3 months. Lesions consisted of intimal atheromata and calcified internal elastica and caused luminal narrowing. The incidence of atherosclerotic lesions was proportional to the vitamin D3 doses. The present experimentally induced lesions had many morphological features resembling those in coronary arteries from human subjects.

Morphological alteration of aortic wall and mitotic cells after complete endothelial loss induced by repeated balloon denudation of swine aorta.

The aortic endothelium in weanling swine was rubbed ten times with an inflated balloon catheter in a repeated balloon denudation. This procedure produced more drastic, extensive and uniform changes in the aortic wall than the commonly used balloon denudation. Sequential alternations of regenerating endothelium, intimal thickening and medial reaction, and the incidence of mitotic cells were studied by scanning and transmission electron microscopy. Complete endothelial loss was confirmed from the descending thoracic aorta to the right femoral artery within 24 hr. By the third day, regenerated endothelium began to cover over the endothelial defect area from the uninjured areas such as the aortic arch and the orifices of branching arteries. Thrombus formation and fibromuscular intimal thickening were observed in the endothelial defect areas by the fifth and seventh days. Three types of mitotic cells, such as endothelial, intimal and medial cells were noted in the aortic wall. Mitotic endothelial and mitotic medial cells were most frequent at Day 3; the latter were closely associated with dead medial cells. Mitotic intimal cells initially appeared at Day 3 and were most frequent at Day 7. Mitotic intimal and mitotic medial cells were frequently present in the aortic wall subjacent to the endothelial defect areas containing interstitial blood components. The large numbers of mitotic aortic cells indicated that endothelial cells give rise to new endothelial cells, intimal cells to new intimal cells, and medial cells to new medial cells.

Ultrastructure of cardiovascular lesions induced by hypervitaminosis D and its withdrawal.

Aortic, coronary and cardiac lesions were induced in swine by a hypervitaminosis D3 diet Lipid-free intimal plaques overlying focally calcified medial or internal elastica occured in the thoracic aorta, pulmonary trunk and coronary artery of swine fed a basal ration supplemented with 250,000 IU of vitamin D3/kg of diet for an induction period of 4 months. Cartilage formation with minimal calcification was initiated in the aortic valve during this period. When such animals were subsequently fed only the basal ration free of excessive vitamin D3 for 3 months, intimal plaques regressed in the aorta and pulmonary trunk but progressed in the coronary artery. The calcific deposits in the medial elastica and internal elastica of all three arteries decreased in size. Atherosclerotic intimal thickening occurred in the main coronary arteries. The most severe lesion occupied 75 p.100 of the lumen area. Multiple intimal plaques were noted in the left atrium and aortic and mitral valves. The thickened intima at these coronary and cardiac sites contained calcified elastica and collagen fibers.

Mitotic structure of aortic intimal cells induced by mechanical injury in swine.

Repair processes of the aortic wall have been studied electron microscopically after removal of the endothelium by an inflated balloon procedure in the thoracic aortae of swine. Initial responses after injury included a thrombotic reaction, the appearance of intimal cells over the denuded surface, and increased mitotic activity of medial cells adjacent to the dead cells by day 3. Cells which engaged in intimal thickening were classified as modified smooth muscle cells throughout the course of this investigation. Mitosis of intimal cells, which was initially observed at day 3, substantially increased at day 7 and decreased by day 14. Mitotic intimal cells contained the same cytoplasmic organelles as interphase modified smooth muscle cells. A detailed description of the paired cisternae as an ultrastructural feature of cell division of modified smooth muscle cells was provided. The paired cisternae were initially observed among the remnants of the nuclear envelope in late prophase; they remained at the periphery of the mitotic apparatus in meta- and anaphase, and finally attached themselves to the nuclear envelope of the daughter cells in late telophase.

Ultrastructure of botryoid sarcoma of the common bile duct.

A case of botryoid sarcoma of the common bile duct in a 4-year-old girl was reported. Electron microscopic examination disclosed that the neoplasm consisted of three types of cells: polygonal, elongated, and small cells. The former two contained moderate to large amounts of poorly developed myofibriles in the cytoplasm with occasional A, I and Z-bands. The small cells contained mitochondria and dilated rough endoplasmic reticulum but few myofilaments. Deposits of glycogen granules were constant components of the neoplastic cells. Mitosis was striking in the small cells. Abnormal multilaminar endoplasmic reticulum was observed in the small cells in the mitotic stage.

Duplication cyst of the duodenum in the adult.

A duodenal duplication cyst in a 45-year-old male is reported. A hypotonic duodenography showed a smooth spherical defect in the descending portion of the duodenum. A fiberoptic duodenoscopy disclosed a smooth well-defined tumor which was located orally from the ampulla of Vater. A retrograde pancreatocholangiography indicated the tumor was not in communication with the pancreatic duct or biliary tract. At operation, a cystic spherical mass, 3 cm in diameter, was located in the posterior wall of the duodenum corresponding to the above described diagnosis. The combination of a greater awareness of this condition as well as improved X-ray and endoscopic techniques has made preoperative diagnosis more accurate.

An ultrastructural study of the Brunner's cyst.

A Brunner's cyst, removed from a 54-year-old woman, was studied by light and electron microscopy. The cyst was spherical, measured 1.5 cm in diameter and was located in the duodenal bulb. Histologically, the cyst, confined in the submucosa, consisted of a single layer of epithelial cells and connective tissue. The epithelial cells were composed of an orderly array of tall columnar cells containing basal round nuclei and fine granular cytoplasm. Ultrastructurally, the epithelial cells contained many secretory granules in the cytoplasm. Each secretory granule was membrane-bound and appeared electron-lucid with a dense core. They were small and sparse around the Golgi apparatus but large and numerous in the apical region. Multinucleated cells were intermingled with cells of the epithelial lining.

Ultrastructural studies of experimental acute pancreatitis.

Acute pancreatitis was studied by electron microscopy after retrograde infusion of either trypsin, and/or beta-glucuronidase into the canine pancreatic duct. Marked changes were induced by the mixture of trypsin and beta-glucuronidase. (1) The acinar cells were initially excavated from the acinar lumen and formed cystic bodies in themselves. The cystic bodies were then disrupted at their marginal membranes, and the acinar cells were filled with a large amount of fibrillar materials which originated from the contents of the cystic bodies. At this time, the luminal margin of the acinar cells completely disappeared. (2) The cellular organellas and the intracellular fibrillar materials in the acinar cells were discharged into the interstitial space through the disrupted basal lamina. Infection in the pancreatic ductal system was considered to play an important role in the pathogenesis of acute hemorrhagic pancreatitis.

Portal hypertension due to hepatic artery-portal vein arteriovenous fistula--a case report.

A case of portal hypertension secondary to traumatic hepatoportal arteriovenous fistula with portal fibrosis was successfully treated by ligation of the afferent hepatic arteries which decreased significantly portal pressure and corrected the abnormal blood inflow to the portal vein via A-V fistula resulting in a recovery of the disturbed liver function. Collateral blood supply from the left hepatic artery into the right hepatic lobe was found to be quite satisfactory after the ligation of the hepatic artery. Hemodynamic data and clinical findings of the present case suggest that the mechanism responsible for the portal hypertension is the inflow block resulting from the interruption of portal venous flow by the inflow of arterial blood via A-V fistula and the subsequent increased blood pressure in portal vein radicals.

Ultrastructural studies of human acute pancreatitis.

The pancreatic tissue from a patient with acute hemorrhagic necrotizing pancreatitis was studied by routine electron microscopic observation. The remarkable change was the destruction of pancreatic acinar units. 1) The acinar lumen was dilated with filling of fibrillar materials, which occasionally contained degenerated cellular components or neutrophils. 2) Accumulations of fibrillar materials were present at the periphery in the acinar cells, especially depositted thickly at the basal portion. 3) In some acinar cells, the accumulation of fibrillar materials occupied the entire acinar cell, accompanied by disappearance of luminal margin, and the intracellular fibrillar materials were mixed with acinar lumen contents. 4) The acinar units which showed above described changes had almost intact basal lamina.