Novel variations and loss of heterozygosity of BRCA2 identified in a dog with mammary tumors

OBJECTIVE: To establish novel polymorphic markers for analysis of loss of heterozygosity (LOH), so as to study the possible involvement of BRCA2 in mammary tumors obtained from dogs. SAMPLE POPULATION: Blood samples, mammary gland specimens, or mammary tumors from 3 tumor-bearing dogs and 10 tumor-free dogs. PROCEDURES: Nucleotide sequence analysis was performed with a DNA autosequencer. Loss of heterozygosity analysis was performed for markers established in the present study. The expression level of canine BRCA2 was quantified by real-time PCR analysis. RESULTS: 3 novel microsatellite markers with high heterozygosity rates (> 50%) were established, and the previously reported marker for canine BRCA2 gene locus was improved. These markers were used for the analysis of DNA from formalin-fixed and paraffin-embedded samples. By use of these markers, LOH in canine BRCA2 was identified as a result of recombination. In mammary tumor DNA that corresponded to the LOH-positive dog, the level of canine BRCA2 expression was decreased compared with that of nonneoplastic mammary gland tissue; the open reading frame contained 4 missense variations, 1 insertion variation, and 1 silent variation, some of which were localized to functional domains. CONCLUSIONS AND CLINICAL RELEVANCE: 3 novel polymorphic markers were developed for LOH analysis of canine BRCA2 and identified a dog with LOH with some variations in the functional domains. These markers could be useful for assessing the relevance of BRCA2 variation in mammary tumors of dogs.     

Seasonal variation of paulownia witches’-broom phytoplasma in paulownia trees and distribution of the disease in the Tohoku district of Japan

Seasonal variation of paulownia witches’-broom (PWB) phytoplasma within different organs (leaves, branch and trunk bark and roots) in paulownia trees was investigated by the amplification of a PWB-specific DNA fragment by the polymerase chain reaction (PCR). In leaf samples, PWB phytoplasma was first detected in June and the incidence gradually increased. On the other hand, the PWB was detected at relatively low incidence in branch bark, trunk bark and roots and the incidence did not change among seasons. A survey of PWB in 27 fields in the Tohoku district of Japan showed that malformed flower buds were observed in paulownia trees in almost all of the fields. PWB-phytoplasma was also detected by PCR from paulownia trees in almost all of the fields in Iwate and Fukushima Prefectures. The frequencies of trees in which phytoplasma was detected by PCR were higher than those in which symptoms were observed. These results indicated that PCR amplification of a PWB-specific DNA fragment is an effective tool for practical diagnose and that PWB is widely distributed in the Tohoku district of Japan.     

Concentrations of trace elements in <Emphasis Type="Italic">Meretrix</Emphasis> spp. (Mollusca: Bivalva) along the coasts of Vietnam

The dominant coastal bivalve in Vietnam, hard clams Meretrix spp., collected from the South Key Economic Zone (SKEZ), the Mekong River Delta, and the Central Coastal Zone (CCZ) were analyzed for 21 trace elements. Comparison of the results from the three regions indicated that levels of most of the trace elements, especially As, Mo, Sn, and Pb, were highest in the samples collected from the CCZ, whereas most of the trace elements were found to be present at relatively low levels in samples from the SKEZ. The high concentrations of these trace elements in the CCZ, a sparsely populated region with less human activity than the other two regions, were believed to have originated from industrial waste produced in a shipyard. Although the trace element concentrations in the bivalves were within safe levels for human-consumption criteria reported by the United State Food and Drug Administration and the European Commission, estimation of cancer risk based on As concentration indicates that the hard clams from the CCZ pose a high risk to consumers. Thus, the industrial waste produced in the less densely populated region might increase the health risk to consumers via the contamination of bivalves commonly used as food.     

Dynamic left ventricular outflow tract obstruction secondary to hypovolemia in a German Shepard dog with splenic hemangiosarcoma

Dynamic left ventricular outflow tract obstruction (DLVOTO) is a common condition in cats and humans. In this case report, a dog is described with DLVOTO secondary to severe intra-abdominal hemorrhage caused by a hemangiosarcoma. The dog was a 9-year-old, 35.7-kg, spayed female German Shepard dog that presented with a history of tachypnea and collapse. A Levine II/VI systolic murmur was present at the heart base. Abdominal ultrasonography revealed a splenic mass and a large amount of ascites. Echocardiography showed a reduced left ventricular diameter and an increased aortic velocity caused by systolic anterior motion (SAM) of the mitral valve apparatus. The heart murmur and the SAM were resolved after treatment including a splenectomy and a blood transfusion.     

Use of live and dead probiotic cells in tilapia Oreochromis niloticus

ABSTRACT:   To investigate the effect of live and dead probiotic cells on the non-specific immune system of tilapia Oreochromis niloticus , probiotics were introduced by feeding either in the form of live or dead cells, or supplying live cells to the rearing water in a closed recirculating system. The probiotics treatment enhanced non-specific immune parameters such as lysozyme activity, migration of neutrophils and plasma bactericidal activity, resulting in improvement of resistance to Edwardsiella tarda infection. Especially, oral administration of live cells seemed to be more effective compared with other probiotic treatments such as oral administration of dead probiotic cells and supply of live probiotic cells to the rearing water. These results indicate that probiotics treatment is promising as an alternative method to antibiotics for disease prevention in aquaculture, and the viability of probiotic bacteria is a key factor to induce more potential effect of probiotics used for fish production.     

Comparison of extractive components in muscle and liver of three Loliginidae squids with those of one Ommastrephidae species

Extractive components were determined for the mantle muscle and liver of three species of Loliginidae squid, Sepioteuthis lessoniana, Loligo bleekeri, and L. edulis, and one species of Ommastrephidae, Todarodes pacificus, as a control. Total free amino acid levels and the major free amino acids in muscle, taurine, proline, glycine, alanine, and arginine, were significantly higher in Loliginidae squids than in T. pacificus. The main nucleotide was adenosine 5′-monophosphate, which did not differ significantly between species. Malate was the organic acid found in muscle in highest concentrations. The muscle of these species contained a large amount of trimethylamine oxide. A large amount of glycine betaine was also detected in the muscle, but showed no large species difference. From these results, the muscle of Loliginidae species is considered to have a much sweeter taste than that of T. pacificus. Compared with muscle, squid liver was characteristic with high contents of taurine, glutamate, bitter amino acids, succinate, propionate, trimethylamine, and glycine betaine, and with low contents of sweet amino acids, arginine, nucleotides, malate, and trimethylamine oxide. These results suggest that squid liver is characterized by a complicated taste containing umami, bitterness, sourness, fishy flavor, and less sweetness.     

Age validation,growth and annual reproductive cycle of chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> off the waters of northern Kyushu and in the East China Sea

The age and growth of chub mackerel Scomber japonicus collected from the East China Sea and the northern waters off Kyushu between June 2000 and June 2001 were determined by observing the otolith surface after dipping it in xylene. The translucent and opaque zones on the otolith surface were identified, and the number of translucent zones was counted. Monthly changes in the frequency of fish with translucent zones on the otolith margin, and in the marginal increments, indicated that the translucent zones were formed between April and June. The seasonal pattern of annulus formation on the otolith became clear by observing the otoliths of fish with known ages, and the otolith formation in wild fish was consistent with that of fish with known ages. The mean gonadosomatic index of male and female fish was high from March to May, and spawning females were observed from mid-March to mid-May. The estimated ages were 1–5 years for males and 1–6 years for females. The von Bertalanffy growth parameters did not significantly differ between male and female. The model was obtained as FL _t=406×{1−exp[−0.372×(t+1.68)]     

Age, growth, and reproductive characteristics of dolphinfish Coryphaena hippurus in the waters off west Kyushu, northern East China Sea

The growth and reproductive characteristics of dolphinfish Coryphaena hippurus collected in the waters off western Kyushu from May 2008 to April 2011 were determined based on scale and otolith readings and gonad histological examinations, respectively. Based on annual increments in scales and daily increments in sagittal otoliths, the von Bertalanffy growth curves in male and females were determined as $ FL_{t} = 1049[1 - \exp \{ - 0.835(t + 6.975 \times 10^{ - 14} )\} ] $ and $ FL_{t} = 938[1 - \exp \{ - 1.029(t + 6.975 \times 10^{ - 14} )\} ] $ , respectively, where FL _t is the mean fork length (mm) at age t. The spawning period was found to last from June to August for dolphinfish, based on an examination of the monthly changes in the gonadosomatic index and histological observations. Therefore, based on the relationship between the fork length and the developmental stage of the testes or ovaries, male and female dolphinfish were found to reach sexual maturity by the following spawning season after hatching in the northern East China Sea.