Principal component analysis uncovers cytomegalovirus-associated NK cell activation in Ph+ leukemia patients treated with dasatinib.

Dasatinib treatment markedly increases the number of large granular lymphocytes (LGLs) in a proportion of Ph+ leukemia patients, which associates with a better prognosis. The lymphocytosis is predominantly observed in cytomegalovirus (CMV)-seropositive patients, yet detectable CMV reactivation exists only in a small fraction of patients. Thus, etiology of the lymphocytosis still remains unclear. Here, we identified NK cells as the dominant LGLs expanding in dasatinib-treated patients, and applied principal component analysis (PCA) to an extensive panel of NK cell markers to explore underlying factors in NK cell activation. PCA displayed phenotypic divergence of NK cells that reflects CMV-associated differentiation and genetic differences, and the divergence was markedly augmented in CMV-seropositive dasatinib-treated patients. Notably, the CMV-associated highly differentiated status of NK cells was already observed at leukemia diagnosis, and was further enhanced after starting dasatinib in virtually all CMV-seropositive patients. Thus, the extensive characterization of NK cells by PCA strongly suggests that CMV is an essential factor in the NK cell activation, which progresses stepwise during leukemia and subsequent dasatinib treatment most likely by subclinical CMV reactivation. This study provides a rationale for the exploitation of CMV-associated NK cell activation for treatment of leukemias.

Six-axis multi-anvil press for high-pressure, high-temperature neutron diffraction experiments.

We developed a six-axis multi-anvil press, ATSUHIME, for high-pressure and high-temperature in situ time-of-flight neutron powder diffraction experiments. The press has six orthogonally oriented hydraulic rams that operate individually to compress a cubic sample assembly. Experiments indicate that the press can generate pressures up to 9.3 GPa and temperatures up to 2000 K using a 6-6-type cell assembly, with available sample volume of about 50 mm(3). Using a 6-8-type cell assembly, the available conditions expand to 16 GPa and 1273 K. Because the six-axis press has no guide blocks, there is sufficient space around the sample to use the aperture for diffraction and place an incident slit, radial collimators, and a neutron imaging camera close to the sample. Combination of the six-axis press and the collimation devices realized high-quality diffraction pattern with no contamination from the heater or the sample container surrounding the sample. This press constitutes a new tool for using neutron diffraction to study the structures of crystals and liquids under high pressures and temperatures.

Integration of linkage and chromosome maps of red clover (Trifolium pratense L.).

Red clover (Trifolium pratense L.) is a forage legume and an allogamous diploid plant (2n = 14; 440 Mb). Here, we examine the 7 prometaphase chromosomes of red clover using fluorescence in situ hybridization (FISH) with ribosomal RNA sequences, pericentromeric and telomeric repeats, as well as bacterial artificial chromosome (BAC) clones. Position of hybridization signals and chromosome condensation patterns were quantified by the help of the chromosome image analysis system ver. 4.0 (CHIAS IV). Fourteen BAC clones belonging to linkage groups (LG) 1-7 hybridized to individual chromosomes 4, 2, 6, 5, 1, 7, and 3, respectively. Quantitative analysis of FISH mapping and chromosome analysis using CHIAS IV allowed us to construct a quantitative idiogram that constitutes the comprehensive chromosome map of red clover. Chromosomal positions of the 26S rDNA locus were detected at a heterozygous locus on chromosome 6 in the variety HR, and polymorphisms of rDNA loci were observed in other varieties, although chromosomal positions of some BAC clones did not vary among HR and other varieties. These results demonstrate chromosomal collinearity among allogamous red clover varieties. This integration of genetic linkage and quantitative chromosome maps should provide valuable insight into allogamous legume genetics.

Avaliação preliminar da utilização de medicamentos empacientes idosos em um hospital da região noroeste paulista / Preliminary assessment in the use of medications by elderly patients in ahospital in northwestern of São Paulo state

Objetivos: Analisar prescrições medicamentosas e patologias em prontuários de idosos atendidos no Hospital Dr. Fernando S/C Ltda na cidade de Mirassol, região Noroeste Paulista. Materiais e Métodos: Estudo do tipo prospectivo com análise de 30 prontuários de pacientes acima de 60 anos, com classificação química, terapêutica e anatômica (ATC), dos medicamentos, verificação de interações medicamentosas pelos softwares Drug Interaction Facts on Disc®, ano de 1999 e Vade-Mécum® 2005-2006, online nos sites www.drugs.com e www.medscape.com e literatura especializada, análise de indicadores de saúde (prescrição) citados pela Organização Mundial de Saúde (OMS) e classificação dos medicamentos em apropriados ou inapropriados para idosos. Resultados:Trezentos e doze medicamentos fizeram parte dos 30 prontuários analisados, com média de 10,4 medicamentos por cada paciente idoso hospitalizado. A prescrição pelo nome genérico, por meio da Denominação Comum Brasileira (DCB), foi feita para 27,23% (n=85) dos medicamentos. Dentre os medicamentos prescritos, 46,79% (n=146) eramantimicrobianos e 76,6% (n=239) injetáveis. Estavam presentes na Relação Municipal de MedicamentosEssenciais (REMUNE) de Mirassol 72,11% (n=225) dos medicamentos presentes nos prontuários. Trinta ecinco dos 312 medicamentos foram considerados inapropriados. Em 70% (n=21) dos prontuários foramdetectadas interações medicamentosas. As patologias mais freqüentes foram aquelas do sistema cardiovascular. Conclusão: A compreensão dos padrões de prescrição é essencial para planejar e estabelecer melhorias nos serviços de assistência à saúde, visando a eficácia no tratamento e segurança do paciente.

Objectives: To analyze drugprescriptions and disorders in medical records of some of the elderlies who are patients at Hospital Dr. Fernando S/C Ltda, in the city of Mirassol, Northwestern of São Paulo State. Materials and Methods: A prospective study analyzing the medical records of 30 patients aged over 60 years including chemical, treatment and anatomical (ATC Portuguese acronym) classifications of the medicine, drug interaction, observation using Drug Interaction Facts Software on Disc ® (1999) and Vade Mecum 2005-2006 softwares. They were available online www.drugs.com and www.medscape.com, and specialized literature; analysis of health indicators (prescription) by World Health Organization (WHO) and classification of appropriate or inappropriate drugs for the elderly. Results: Three hundred and twelve drugs appeared on the 30 reviewed medical records, with an average of 10.4 medications per hospitalized elderly patient. Prescription with generic name, taken at the Brazilian Common Denomination (DCB Portuguese acronym), was made for 27.23% (n = 85) of the drugs. Among prescribed drugs, 46.79% (n = 146) were antimicrobial agents and 76.6% (n = 239) were of injectable-drugs use. In 72.11% (n = 225) of the drugs found on the medical records were listed at the Mirassol-Municipal Register of Essential Medicines (REMUNE Portuguese acronym). Thirty-five of 312 medications were considered inappropriate. In 70% (n = 21) of medical records, drug interactions were detected. The most frequent diagnosed disorders were related to the cardiovascular system. Conclusion: The understanding of medicine prescribing patterns is essential for planning and implementing better services in the health care work, aiming efficacy of the treatment and the patient´s safety.

Ectopic ossification with haematopoietic bone marrow in the heart valves of a crossbred heavy horse.

Ectopic bone formation in the left atrioventricular valves and cardiac fibroskeleton, with systemic circulatory disturbance, is reported in a 4-year-old crossbred heavy horse. Microscopically, there was fibrosis, chondral metaplasia and mature bone, with bone marrow within the left atrioventricular cusps and in the annuli of the aortic and right atrioventricular valves.

Exposure to aflatoxins in Japan: risk assessment for aflatoxin B1.

The intake of total aflatoxins (AFT) and aflatoxin B(1) (AFB(1)) from food in Japan was estimated from AFT and AFB(1) concentration and frequency data in 24 foods (884 samples) from a 3-year retail market survey from the summer of 2004 to the winter of 2006, and by food consumption data from the National Health and Nutrition Survey performed in 2005. The AFT and AFB(1) survey revealed that peanut, peanut products, cocoa, chocolate, pistachio, white pepper, red pepper, almond, job's tears, buckwheat and corn grits are considered to be contributors of AFT (or AFB(1)) intake in Japan (maximum AFB(1) (AFT) levels ranged from 0.21 to 28.0 microg kg(-1) (from 0.21 to 9.0 microg kg(-1))) in AFT-contaminated food. A probabilistic approach using the Monte Carlo method was carried out to simulate an estimate of the AFT (or AFB(1)) intake distributions in each age group in Japan. In this study, AFB(1) intake ranged from 0.003 to 0.004 ng kg(-1) body weight day(-1) (from lower to upper limits), and the potential risk for cancer using a formula devised by the Joint Food and Agricultural Organization/World Health Organization (FAO/WHO) Expert Committee on Food Additives (JECFA) was estimated at 0.00004-0.00005 person/year/100,000 persons, even though this was in the higher levels (95.0th percentile) of the consumer population. The results suggest that the current dietary intake of AFB(1) in Japan has no appreciable effect on health.

Collagen content and architecture of the puboischiofemoralis muscle in male chicks and broilers with different growth rates on various nutritional planes.

1. Various growth rates of chickens were induced with different nutritional regimes, and the collagen content and architecture of the medial part of the puboischiofemoralis muscle were compared among 21-d-old chicks and 80- or 95-d-old broilers. 2. The percentage muscle weight relative to live weight increased from chicks to 80-d-old broilers and the 95-d-old broilers attained the largest percentage. An inter-relationship of the percentage muscle weight and the growth rates of birds could not be determined. 3. Collagen concentration was related to the growth rates for the first 21 d post hatching and maintained the same level during the later stages up to 80 d. The 95-d-old broilers, that were subjected to early rapid growth followed by restricted later growth, had the highest collagen content. 4. On SEM (Scanning Electron Microscopy) photographs, endomysial honeycombs were small and encircled by perimysia of a collagen network with small mesh size. Thin and thick perimysia were distinguished and the expanded portion of thick perimysia was also observed. Generally, the perimysia were made up of rough collagen tissue where fatty tissue developed, especially in the broilers. 5. Perimysial collagen fibres with mainly transverse striation were divided into two fundamental types, wide collagen platelets and narrow cords. With growth from the chick to broiler stage, features of the collagen fibres did not change regardless of expansion of the thick perimysia. Endomysia increased slightly from thin to thick meshwork as growth progressed. However, the collagen architecture of the muscle in broilers did not change under different nutritional regimes. 6. In conclusion, the puboischiofemoralis muscle of chickens develops relative to live weight when later growth is limited in broilers, but the collagen architecture is not affected by the different growth rates.

Collagen content and architecture of the Iliotibialis lateralis muscle in male chicks and broilers with different growth rates fed on different nutritional planes.

1. Varying growth rates in chickens were induced by different nutritional regimes. The collagen content and architecture of iliotibialis lateralis (ITL) muscle were compared among 21-d-old chick types and broiler types at 80 or 95 d of age. 2. Relative size of ITL muscle was greater in the rapid growing (1.16% of live weight) than the slow growing chicks (1.02% of live weight). The 80-d-old broilers with a compensatory growth phase after an earlier slow growth period produced ITL muscle at 1.65-1.69% of live weight. The ITL muscle in 80- and 95-d-old broilers with restricted later growth after an earlier rapid growth period was 1.29 and 1.49% of live weight, respectively. 3. Collagen content of ITL muscle did not differ between chick types and also among the broiler types. However, collagen concentration decreased from 6.00-6.51 mg/g in the chicks to 3.33-4.00 mg/g in the broilers. 4. Thick and thin perimysia and honeycomb endomysia were viewed by scanning electron microscope (SEM) photography. In the perimysia, a central wide layer of longitudinal collagen fibres and peripheral narrow band of transverse fibres were distinguished. Collagen baskets of adipocytes were observed in the perimysia. 5. Perimysial collagen fibres markedly increased in number and formed a larger fibre cluster during growth from chicks to broilers. Endomysia changed from thin to thicker meshwork with growth. However, the collagen architecture of the muscle in broilers did not change under different nutritional regimes. 6. In conclusion, ITL muscle of chicken develops optimally when body growth is enhanced, but the collagen content and architecture in broilers are not affected by different growth processes.

Aflatoxin and ochratoxin A contamination of retail foods and intake of these mycotoxins in Japan.

A survey was undertaken of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), ochratoxin A (OTA), and fumonisin B1 (FB1), B2 (FB2) and B3 (FB3) contamination of various retail foods in Japan during 2004-05. The mycotoxins were analysed by high-performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS) or high-performance thin-layer chromatography (HPTLC). Aflatoxins (AFs) were detected in ten of 21 peanut butter and in 22 of 44 bitter chocolate samples; the highest level of AFB1, 2.59 microg kg(-1), was found in peanut butter. Aflatoxin contamination was not observed in corn products (n = 55), corn (n = 110), peanuts (n = 120), buckwheat flour (n = 23), dried buckwheat noodles (n = 59), rice (n = 83) or sesame oil (n = 20). OTA was detected in 120 out of 192 samples of oatmeal, wheat flour, rye, buckwheat flour, raw coffee, roasted coffee, raisin, beer, wine and bitter chocolate, but not in rice or corn products. OTA levels in the positive samples were below 13 microg kg(-1). AFs and OTA intakes through the consumption of foods containing cacao were estimated using the data for mycotoxin contamination in bitter chocolate and those for the consumption of foods containing cacao in Japan.

Three-dimensional observation of connective tissue of bovine masseter muscle under concentrate- and roughage-fed conditions by using immunohistochemical/confocal laser-scanning microscopic methods.

We investigated changes in connective tissue components of masseter (MA) muscle in Japanese black heifers (n= 6) in concentrate- and roughage-fed groups (groups C and R, respectively). Body weight, at slaughter, of experimental heifers in group C (272.3 +/- 22.3 kg) was higher (P < 0.05) than that of group R (213.8 +/- 27.5 kg). However, muscle weight and myofiber diameter (superficial and deep layers) of MA muscle did not differ between groups C and R. In contrast, total mastication duration of group R was longer (P < 0.05) than that of group C. MA muscle of groups C and R was composed only of type I myofiber. Using immunohistochemical/confocal laser-scanning microscopy, type I collagen was observed mainly in perimysium, and type V and VI collagen were observed in perimysium and endomysium of both groups. Type IV collagen and laminin were observed only in the endomysium in both groups. However, type III collagen and fibronectin were strongly apparent in the perimysium and endomysium in group R. Connective tissue components in the perimysium of groups C and R were observed to form plate-shaped layers. On the other hand, honeycomb-shaped connective tissue components were seen in the endomysium-surrounded muscle fibers. In particular, fibronectin was strongly observed in the perimysium and endomysium in group R. These results indicate that there are different developmental changes among connective tissue components in MA muscle in response to mastication. The immunohistochemical/confocal laser-scanning microscopic method is useful to investigate the structural relationship among connective tissue components in skeletal muscle.

Histochemical properties and collagen architecture of M. iliotibialis lateralis and M. puboischiofemoralis in male broilers with different growth rates induced by feeding at different planes of nutrition.

1. The histochemical properties and the collagen content and architecture of the iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles were assessed in Red Cornish x New Hampshire cockerels reared on a high nutrient plane for 80 d (H80d), or a low nutrient plane for 80 d (L80d) or 95 d (L95d). 2. Final live weights were 3410 g in H80d, 2810 g in L80d and 3467 g in L95d. Both ITL and PIF muscle weights were lowest in L80d and did not differ between H80d and L95d. 3. ITL muscle was composed of fast-twitch myofibres such as IIA (high reduced nicotinamide adenine dinucleotide dehydrogenase, NADH-DH activity), IIB (low NADH-DH activity) and IIC (intermediate NADH-DH activity). The high percentage of type IIB myofibres in H80d (76.6%) and L95d (76.2%) birds were reflected in low percentages of type IIC myofibres (12.2%) in H80d birds and type IIA myofibres (8.2%) in L95d birds. Percentages of IIA, IIB and IIC myofibres in L80d cockerels were 12.4, 69.8 and 17.6%, respectively. 4. The myofibres in PIF muscle were divided into two basic types, I and IIA, and a transitional form (I-tr) from IIA to I. In the caudal region, all myofibres in H80d and L95d cockerels were type I but in L80d cockerels 15% of myofibres were categorised as type I-tr. In the cranial region, the great majority (52 to 63%) of myofibres were type IIA. Type I myofibres occurred at a higher percentage in H80d (30.5%) than L95d (21.8%) and type I-tr in L95d (15.7%) than H80d (7.3%) and L80d (11.5%). 5. The total amount of collagen was higher in ITL than PIF muscle in every bird group. In both muscles the highest collagen content was in L95d cockerels but the content did not differ between H80d and L80d birds. The thickness of thick and thin perimysia increased with muscle size. The circular collagen fibre in the thick perimysium was larger in ITL (6.1 to 7.0 microm) than PIF (3.7 to 3.8 microm) muscle but did not differ among the bird groups. 6. From these results, it was concluded that feeding on a high nutritional plane promotes growth of the thigh muscles, with accompanying enlargement of the perimysial thickness, no increase in collagen content and various changes of histochemical properties.

Age-related changes in the intramuscular distribution of melanocytes in the Silky fowl.

1. The characteristics of melanocyte distribution in skeletal muscles in the Silky fowl were investigated in association with growth. 2. Pectoralis (PT) and iliotibialis lateralis (ITL) muscles from 1-, 3-, 5-, 10-, 20- and 30-week-old Silky males were weighed and collagen type I was detected in frozen sections immunohistochemically. 3. Melanocytes were observed in the collagen type I-immunopositive endomysium and perimysium in both muscles. 4. Image analysis indicated that the total area occupied by melanocytes in histological sections sharply decreased from 0.61% to 0.16% in PT muscle and from 1.67% to 0.33% in ITL muscle at 1 to 3 weeks, and then gradually decreased. The melanocyte area was larger in ITL muscle than in PT muscle until 10 weeks of age. 5. We concluded that the proportion of intramuscular melanocytes in the Silky fowl differs between types of muscles in the early stages of development, and it decreases with growth.

Effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle of male broilers.

1. The effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle were evaluated using male broilers of Red Cornish x New Hampshire stock, reared on diets of high nutritional value for up to 80 d (H80d) and low nutritional value for up to 80 d (L80d, same age as H80d) or 95 d (L95d, same body weight as H80d). 2. The total live weight and the weight of pectoralis muscle were lower in L80d than in both H80d and L95d. The muscle weight as a percentage of live weight was 8.7% in L80d, 10.7% in H80d and 11.5% in L95d. 3. Pectoralis muscle was composed only of type IIB myofibres and showed no differences in myofibre type composition among the chicken groups. The largest diameter of type IIB myofibres was observed in L95d, followed by H80d and the smallest in L80d. 4. The total amount of intramuscular collagen did not differ among the chicken groups (1.92 to 1.99 mg/g). Types I and III collagens were immunohistochemically detected in both the perimysia and endomysia. The thin perimysia around the primary myofibre fascicles showed larger width in H80d than L80d and L95d, and also the thick perimysia around the secondary fascicles in H80d than L80d. 5. The collagen structure of the perimysium was most developed in H80d, followed by L95d and on the least in L80d. The development of perimysial collagen fibres could be enhanced by a rapid growth rate of the muscle induced by high nutritional level and depressed by a slow growth rate with low nutritional foods. 6. The endomysial collagen architecture was observed as a felt-like tissue of the fibril bundles with many slits. The thinnest endomysial wall was observed in L80d, followed by H80d and the thickest in L95d. 7. From these results, it was indicated that foods of high nutritional value could enhance growth of the pectoralis muscle of broilers, and this is accompanied by hypertrophy of the type IIB myofibres and development of the perimysial collagen architecture.

[Intraoperative frozen section diiagnosis of small peripheral lung adenocarcinoma for limited operation].

Sixty-two cases of peripheral lung adenocarcinoma 2 cm or less in diameter were diagnosed as type A, B, or C according to Noguchi classifications by intraoperative frozen section diagnosis. Four of 19 type A were changed finally to type B by postoperative pathologic examination. Likewise 10 of 34 type B were changed finally to 1 type A, 8 type C, and 1 type D. Three of 9 type C were changed finally to 2 type B and 1 type D. Frozen section diagnosis for small peripheral lung adenocarcinoma has some special problems and limitations. Only cases of type A diagnosed by frozen section diagnosis should be permitted to undergo limited surgical resection.

[Clinical experience with aortic connector system].

From April to December 2002, 40 patients underwent coronary artery bypass grafting (CABG) using the St. Jude Medical (Minneapolis) Symmetry bypass system (aortic connector system: ACS). 59 proximal anastomoses (51 saphenous vein grafts, 8 radial artery grafts) were performed with the ACS. One saphenous vein graft occluded during operation. Postoperative evaluation of the anastomotic patency was carried out by angiography in 45 grafts. Five of the saphenous vein grafts were occluded (5/38). One patient who was shock state before operation presented with postoperative unconsciousness. Another patient died at 8th postoperative day caused by ventricular fibrillation. We conclude that the ACS produces a simple, quick way of performing the proximal anastomosis without the need for clamping the aorta, allows reducing risk of embolization by aortic manipulation. However, it is necessary to discuss sufficiently using the ACS, because the graft patency with the ACS is lower than with standard suturing technique.

Occurrence of aflatoxin M1 in domestic milk in Japan during the winter season.

A total of 208 samples of commercial pasteurized milk gathered from retail outlets across Japan during the winter season were analysed for aflatoxin M1 (AFM1). Japan was divided into 11 regions from north to south, and nine to 45 milk samples from each region were randomly purchased between December 2001 and February 2002. Each milk sample was cleaned up by an immunoaffinity column, and AFM1 was quantified by liquid chromatography with fluorescence detection in four independent laboratories. The limit of detection of the method was 0.001 microg kg(-1). The identity of the putative AFM1 in milk sample was confirmed by the formation of AFM1 hemi-acetal with trifluoroacetic acid. Based on the results obtained with spiked samples (0.05 microg AFM1 kg(-1)), the mean recovery was 91.4%, the relative standard deviation for repeatability was 4.6%, and the relative standard deviation for reproducibility was 8.0% among four independent laboratories. AFM1 was detected in 207 (99.5%) of 208 milk samples at 0.001-0.029 microg kg(-1), with a mean of 0.009 microg kg(-1) and a 90th percentile of 0.014 microg kg(-1). No significant difference of the level of AFM1 contamination was observed among the regions.

Developmental states of the collagen content, distribution and architecture in the pectoralis, iliotibialis lateralis and puboischiofemoralis muscles of male Red Cornish x New Hampshire and normal broilers.

1. Developmental states of the collagen content, distribution and architecture in the pectoralis (PT), iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles of male Red Cornish x New Hampshire (RN, 80 d, body weight 2.9 kg) and normal (3.1 kg) broilers were evaluated. 2. In PT muscle the total amount of collagen was significantly greater in RN broilers (3.33 mg/g) than in normal ones (1.71 mg/g). This higher collagen content in RN broilers was based mainly on the closer mesh sizes of endomysial honeycomb. The collagen structures in the perimysia also differed between broiler types, when more collagen fibres were observed in RN broilers. 3. ITL muscle contained total collagen of 4.10 to 5.00 mg/g. Types I and III collagens were distributed on the perimysia at higher percentages in RN broilers (31.6%, 37.2%) than normal (15.6%, 30.8%), respectively. The thick bands of tough collagen fibres characteristic of ITL muscle perimysium in cockerels had not yet developed in these broilers. 4. Total collagen was 4.63 to 6.29 mg/g in PIF material with fascia. In PIF muscle the perimysial collagen fibres had not yet attained their full growth but consisted of densely packed fibrils. PIF muscle was characterised by the earlier maturing collagen structure. 5. These results show that a perimysial collagen structure in broilers is still in an undeveloped state. It is supposed that tenderness of broiler meat is attributed mainly to characteristics of the collagen distribution, in which the majority of types I and III collagens is distributed on the closer mesh of endomysial honeycomb.