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1.
Opt Express ; 31(15): 24045-24053, 2023 Jul 17.
Article in English | MEDLINE | ID: mdl-37475241

ABSTRACT

Image-based deep learning (IBDL) is an advanced technique for predicting the surface irradiation conditions of laser surface processing technology. In pulsed-laser surface processing techniques, the number of superimposed laser shots is one of the fundamental and essential parameters that should be optimized for each material. Our primary research aims to build an adequate dataset using laser-irradiated surface images and to successfully predict the number of superimposed shots using the pre-trained deep convolutional neural network (CNN) models. First, the laser shot experiments were performed on copper targets using a nanosecond YAG laser with a wavelength of 532 nm. Then, the training data were obtained with the different superimposed shots of 1 to 1024 in powers of 2. After that, we used several pre-trained deep CNN models to predict the number of superimposed laser shots. Based on the dataset with 1936 images, VGG16 shows a high validation accuracy, higher sensitivity, and more than 99% precision than other deep CNN models. Utilizing the VGG16 model with high sensitivity could positively impact the industries' time, efficiency, and overall production.

2.
Lett Appl Microbiol ; 74(1): 27-31, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34608644

ABSTRACT

Rhodococcus equi was isolated from the gastrointestinal contents of earthworms (family Megascolecidae) and their surrounding soil collected from pastures of two horse-breeding farms in Aomori Prefecture, outdoor pig pens, forest in Towada campus, orange groves and forest where wild boars (Sus scrofa) are established in Tanabe, Wakayama Prefecture. The number of R. equi in the lower gastrointestinal contents of 23 earthworms collected from our campus was significantly larger than that of the upper gastrointestinal content. The mean numbers of R. equi from the gastrointestinal contents of earthworms collected from the various places were 2·3-fold to 39·7-fold more than those of the surrounding soil samples. In all, 1771 isolates from the earthworms and 489 isolates from the soil samples were tested for the presence of vapA and vapB genes using polymerase chain reaction. At the horse-breeding farm N, 9 of the 109 isolates (8·3%) from the earthworms and 7 of the 106 isolates (6·6%) from the soil samples were positive for the vapA gene. At the University's forest, one of the 250 isolates (0·4%) from the gastrointestinal contents of the earthworm was positive for the vapB gene. These results revealed that R. equi can be found in significant quantities in the gastrointestinal contents of earthworms, suggesting that they act as an accumulator of R. equi in the soil environment and as a source or reservoir of animal infection.


Subject(s)
Actinomycetales Infections , Horse Diseases , Oligochaeta , Rhodococcus equi , Animals , Gastrointestinal Contents , Horses , Soil Microbiology
3.
Lett Appl Microbiol ; 75(4): 908-912, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35707941

ABSTRACT

Clinical samples from 123 foals with suspected rhodococcosis submitted to the Veterinary Microbiological Diagnostic Centre of the Faculty of Veterinary Medicine between 1993 and 2006 were tested for the presence of the virulence gene vapA. Of the 123 samples, 120 were vapA-positive and 3 vapA-negative Rhodococcus equi were isolated. The 120 vapA-positive R. equi were isolated from 70 tracheal wash, 19 lung tissues, 7 lymph nodes, 6 synovial fluids, 13 abscesses or pus and single isolates from the uterus, gut, cerebrospinal fluid, abdomen fluid and faeces. Of the 120 isolates, 46 were from Dutch warmblood horses, 23 from Friesian horses, 14 from Trotters, 4 from Holsteiners, 3 from Arab breed, 2 from ponies, 1 from a Welsh pony and 27 from undefined breed horses. Using plasmid profile analysis of the 120 isolates, 117 isolates contained the 85-kb type I plasmid, 2 contained the 87-kb type I plasmid and 1 contained the novel 52-kb non-mobilizable virulence plasmid reported recently. These results showed that the virulent R. equi strains harbouring a virulence plasmid of 85-kb type I or 87-kb type I, which have been detected in clinical isolates from five European countries, are widespread in the Netherlands. This is the first report of plasmid types of clinical R. equi isolates in the Netherlands.


Subject(s)
Actinomycetales Infections , Horse Diseases , Rhodococcus equi , Actinomycetales Infections/epidemiology , Actinomycetales Infections/microbiology , Actinomycetales Infections/veterinary , Animals , Bacterial Proteins/genetics , Female , Horse Diseases/epidemiology , Horses/genetics , Netherlands , Plasmids/genetics , Rhodococcus equi/genetics , Soil Microbiology , Virulence/genetics , Virulence Factors/genetics
4.
Lett Appl Microbiol ; 71(6): 679-683, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32920889

ABSTRACT

Rhodococcus equi emerged as a zoonotic pathogen of human immunodeficiency virus-infected patients over the last three decades. Two virulence plasmid types of R. equi, pVAPA and pVAPB associated with equine and porcine isolates, have been recognized, and more recently, pVAPN, a novel host-associated virulence plasmid in R. equi, was found in bovine and caprine isolates. We reinvestigated 39 previously reported isolates of R. equi from patients with and without acquired immunodeficiency syndrome (AIDS) by detecting vapA, vapB and vapN using PCR and plasmid profiling. After excluding one isolate that could not be cultured from frozen storage, eight isolates carried a virulence plasmid encoding vapA (pVAPA), 10 carried a virulence plasmid encoding vapB (pVAPB), seven carried a virulence plasmid encoding vapN (pVAPN) and 13 were negative for those genes. Of the 29 isolates from patients with AIDS, 7, 10 and 5 harboured pVAPA, pVAPB and pVAPN respectively. Among nine isolates from patients without AIDS, one and two harboured pVAPA and pVAPN respectively. This study demonstrated that pVAPN-positive R. equi existed in human isolates before 1994 and reaffirmed that equine-associated pVAPA-positive, porcine-associated pVAPB-positive and bovine- or caprine-associated pVAPN-positive R. equi are widely spread globally. Because domestic animals might be major sources of human infection, further research is needed to reveal the prevalence of pVAPN-positive R. equi infection in cattle and goats.


Subject(s)
AIDS-Related Opportunistic Infections/microbiology , Acquired Immunodeficiency Syndrome/complications , Actinomycetales Infections/microbiology , Rhodococcus equi/pathogenicity , Acquired Immunodeficiency Syndrome/virology , Actinomycetales Infections/etiology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , HIV/physiology , Humans , Plasmids/genetics , Plasmids/metabolism , Polymerase Chain Reaction , Rhodococcus equi/classification , Rhodococcus equi/genetics , Rhodococcus equi/metabolism , Virulence
5.
Gene Ther ; 25(1): 20-26, 2018 01.
Article in English | MEDLINE | ID: mdl-29057994

ABSTRACT

Enhanced afferent excitability is considered to be an important pathophysiological basis of interstitial cystitis/bladder pain syndrome (IC/BPS). In addition, transient receptor potential vanilloid-1 (TRPV1) receptors are known to be involved in afferent sensitization. Animals with hydrogen peroxide (HP)-induced cystitis have been used as a model exhibiting pathologic characteristics of chronic inflammatory condition of the bladder. This study investigated the effect of gene therapy with replication-defective herpes simplex virus (HSV) vectors encoding poreless TRPV1 (PL) or protein phosphatase 1 α (PP1α), a negative regulator of TRPV1, using a HP-induced rat model of cystitis. HSV vectors encoding green fluorescent protein, PL or PP1α were inoculated into the bladder wall of female rats. After 1 week, 1% HP or normal saline was administered into the bladder, and the evaluations were performed 2 weeks after viral inoculation. In HP-induced cystitis rats, gene delivery of PL or PP1α decreased pain behavior as well as a reduction in the intercontraction interval. Also, both treatments reduced nerve growth factor expression in the bladder mucosa, reduced bladder inflammation characterized by infiltration of inflammatory cells and increased bladder weight. Taken together, HSV-mediated gene therapy targeting TRPV1 receptors could be effective for the treatment of IC/BPS.


Subject(s)
Cystitis/chemically induced , Cystitis/therapy , Genetic Therapy/methods , Genetic Vectors , Hydrogen Peroxide/toxicity , Protein Phosphatase 1/genetics , Simplexvirus/genetics , TRPV Cation Channels/genetics , Animals , Cystitis/enzymology , Cystitis/metabolism , Defective Viruses/genetics , Disease Models, Animal , Female , Gene Expression , Green Fluorescent Proteins/genetics , Organ Size , Rats , Urinary Bladder/drug effects , Urinary Bladder/pathology
6.
Lett Appl Microbiol ; 61(3): 306-10, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26103784

ABSTRACT

UNLABELLED: The virulence-plasmid profile of Rhodococcus equi strains isolated from Suidae and humans is similar. Recent evidence suggests that the consumption of pork products contaminated with faeces might be a potential source of R. equi infections in humans, mainly to patients with rhodococcosis without history of contact with pigs or pig farms. This study investigated the virulence-associated genes (vapA and vapB) and plasmid profiles of R. equi among the 150 samples of small intestinal content obtained from slaughtered pigs. In addition, all samples were subjected to microbiological culture in conventional sheep blood agar and CAZ-NB, TCP and TVP selective media. A total of 40 (26·7%) of the samples recovered R. equi, with two samples recovering isolates harbouring the VapB type 8 plasmid. Among the 150 pigs sampled herein, CAZ-NB was considered the best selective medium for the isolation of R. equi from faeces. Our results provide evidence that the contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, this study is the first description of R. equi strains carrying the VapB plasmid in the gut of pigs. SIGNIFICANCE AND IMPACT OF THE STUDY: Intermediately virulent (VapB) is a common plasmid-type harboured by R. equi isolated from pigs and humans with AIDS. Curiously, humans with rhodococcosis usually have no history of contact with pigs or pig farms. Virulence-plasmid profile of 40 R. equi isolated among 150 small intestine content samples from pigs revelled two carrying isolates with the VapB type-8 plasmids. Moreover, comparison of three selective culture media shows that CAZ-NB was the best. Our results provide evidence that contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, R. equi carrying VapB type-8 plasmids types are described for the first time in the gut of the pig.


Subject(s)
Actinomycetales Infections/microbiology , Bacterial Proteins/genetics , Culture Media , DNA-Binding Proteins/genetics , Food Microbiology , Membrane Glycoproteins/genetics , Red Meat/microbiology , Rhodococcus equi/isolation & purification , Abattoirs , Animals , Brazil , Feces/microbiology , Humans , Intestine, Small/microbiology , Plasmids/genetics , Plasmids/isolation & purification , Rhodococcus equi/genetics , Swine , Swine Diseases/microbiology , Virulence/genetics , Virulence Factors/genetics
8.
Transbound Emerg Dis ; 65(2): 321-326, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29226632

ABSTRACT

Rhodococcus equi is a well-recognized Gram-positive intracellular facultative bacterium that is opportunistic in nature, which causes pyogranulomatous infections in humans and multiple host animals. The pathogenicity of the microorganism has been attributed to the presence of plasmid-encoded virulence-associated proteins (Vap). To date, three host-associated virulence plasmid types of R. equi have been identified as follows: the circular pVAPA and pVAPB, related, respectively, to equine and porcine isolates, and a recently described linear pVAPN plasmid associated with bovine strains, although these three types are found in human isolates. Recent phylogenomic studies support the evidence that human R. equi infection is zoonotically acquired. Nevertheless, data regarding distribution and prevalence of the host-adapted virulence plasmid types of R. equi isolated from meat animals are scarce or unnoticed. Here, the three host-associated virulence plasmid types (pVAPA, pVAPB, and pVAPN) were investigated in 154 R. equi isolates recovered from lymph nodes of cattle with lymphadenitis (n = 31), faeces of cattle without enteric signs (n = 49), as well as different clinical specimens from human patients (n = 74). The analysis of virulence profile of 74 R. equi from humans revealed six (8.1%) isolates pVAPB (type 8), two (2.7%) pVAPN, and one (1.3%) pVAPB (type 11), all of which were from lung samples from people living with HIV/AIDS. From the lymph node samples of cattle, 41.9% (13 of 31) isolates revealed pVAPN type, whereas all isolates from faecal samples were negative for three host-associated types. Here, recently described bovine-associated pVAPN type was detected in R. equi isolates recovered from the lungs of people living with HIV/AIDS and lymph nodes from slaughtered cattle intended for human consumption; a finding that represents a public health concern, mainly in countries where undercooked or raw meat are traditionally consumed.


Subject(s)
Actinomycetales Infections/veterinary , Bacterial Proteins/genetics , Cattle Diseases/microbiology , HIV Infections/microbiology , Lung Diseases/microbiology , Lymph Nodes/microbiology , Rhodococcus equi/isolation & purification , Virulence Factors/genetics , Actinomycetales Infections/microbiology , Animals , Cattle , DNA, Bacterial/genetics , Feces , Humans , Plasmids/genetics , Prevalence , Rhodococcus equi/genetics
9.
J Clin Invest ; 104(7): 925-35, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10510333

ABSTRACT

Angiotensin II (Ang II) is a potent vasopressor peptide that interacts with 2 major receptor isoforms - AT1 and AT2. Although blood pressure is increased in AT2 knockout mice, the underlying mechanisms remain undefined because of the low levels of expression of AT2 in the vasculature. Here we overexpressed AT2 in vascular smooth muscle (VSM) cells in transgenic (TG) mice. Aortic AT1 was not affected by overexpression of AT2. Chronic infusion of Ang II into AT2-TG mice completely abolished the AT1-mediated pressor effect, which was blocked by inhibitors of bradykinin type 2 receptor (icatibant) and nitric oxide (NO) synthase (L-NAME). Aortic explants from TG mice showed greatly increased cGMP production and diminished Ang II-induced vascular constriction. Removal of endothelium or treatment with icatibant and L-NAME abolished these AT2-mediated effects. AT2 blocked the amiloride-sensitive Na(+)/H(+) exchanger, promoting intracellular acidosis in VSM cells and activating kininogenases. The resulting enhancement of aortic kinin formation in TG mice was not affected by removal of endothelium. Our results suggest that AT2 in aortic VSM cells stimulates the production of bradykinin, which stimulates the NO/cGMP system in a paracrine manner to promote vasodilation. Selective stimulation of AT2 in the presence of AT1 antagonists is predicted to have a beneficial clinical effect in controlling blood pressure.


Subject(s)
Aorta/physiology , Kinins/physiology , Muscle, Smooth, Vascular/physiology , Receptors, Angiotensin/physiology , Tunica Media/physiology , Vasodilation/physiology , Actins/genetics , Amiloride/pharmacology , Angiotensin II/pharmacology , Animals , Blood Pressure/physiology , Bradykinin/analogs & derivatives , Bradykinin/pharmacology , Bradykinin/physiology , Bradykinin Receptor Antagonists , Cell Membrane/physiology , Cyclic GMP/metabolism , Endothelium, Vascular/drug effects , Endothelium, Vascular/physiology , Imidazoles/pharmacology , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Muscle, Smooth, Vascular/drug effects , NG-Nitroarginine Methyl Ester , Promoter Regions, Genetic , Pyridines/pharmacology , Receptor, Angiotensin, Type 1 , Receptor, Angiotensin, Type 2 , Receptors, Angiotensin/deficiency , Receptors, Angiotensin/genetics , Recombinant Fusion Proteins/metabolism , Vasoconstriction , Vasodilation/drug effects
10.
Res Vet Sci ; 83(3): 311-7, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17360011

ABSTRACT

Pulsotypes of VapA positive Rhodococcus equi isolated from foals and soil on a farm in Germany were characterized on the basis of nasal and tracheal samples simultaneously collected in 2003 from 217 foals with sonographic evidence of pneumonia or pulmonary abscesses. Of the 217 double samples, R. equi was isolated in 118 (54%) of the tracheal samples and in 52 of the nasal swab samples (24%) (P<0.001). Furthermore, 37 and 55 isolates were also randomly selected from nasal swabs and the tracheal samples, respectively, and further processed to determine the presence of VapA by colony blot enzyme-linked immunosorbent assay. This method showed that 26 (68%) of the nasal swabs and 40 (73%) of the tracheal samples were VapA-positive. R. equi was isolated from 56 (87%) of the 64 soil samples taken from the paddocks and stables in March and from 17 (68%) of the 25 samples taken in July of 2003. Three (21%) of these randomly selected 14 isolates from March and 13 (81%) of the 16 from July were VapA-positive. The VapA positive isolates from foals and soil were genotyped by plasmid profiling, restriction fragment length polymorphism analysis and pulsed-field gel electrophoresis. Of the 83 isolates, 80 contained an 85-kb type I plasmid and three contained an 87-kb type I plasmid. Pulsed-field gel electrophoresis yielded four distinct VspI profiles dividing 83 isolates into three major (A1, 51; D, 14; and 11 isolates) and three minor (C, 4; A3, 2; and A2, 1 isolates) groups. These results suggest that the majority of foals were exposed to and infected with three pulsotypes of VapA positive R. equi containing an 85-kb type I plasmid.


Subject(s)
Actinomycetales Infections/veterinary , Horse Diseases/microbiology , Lung Abscess/veterinary , Pneumonia/veterinary , Rhodococcus equi/genetics , Soil Microbiology , Actinomycetales Infections/microbiology , Animals , Breeding , Genotype , Germany , Horses , Lung Abscess/microbiology , Pneumonia/microbiology , Virulence Factors/genetics
11.
Vet Rec ; 161(10): 342-6, 2007 Sep 08.
Article in English | MEDLINE | ID: mdl-17827474

ABSTRACT

An eight-year-old thoroughbred mare with no previous history of illness aborted a fetus at 196 days of gestation, and its internal tissues were examined immunohistologically and bacteriologically. The placenta was not examined, but specimens of the intrauterine fluids and the dam's faeces were collected four days after the abortion and examined bacteriologically. No significant histological lesions were found in the fetus but the amnion and the umbilical cord were oedematous and had petechial haemorrhages. Rhodococcus equi was isolated in pure culture from the lung, heart and stomach contents of the fetus and from an intrauterine specimen and faeces of the dam. The anti-R equi antibody titre of the mare was high after the abortion. The diagnosis was confirmed in the lung of the fetus by immunohistochemical staining with R equi-specific antibodies. Isolates from the fetus and mare were identified as avirulent R equi by pcr and the mouse pathogenicity test. The avirulent isolates were characterised by pulsed-field gel electrophoresis, which yielded only one VspI profile in all the isolates from the fetus and its dam.


Subject(s)
Abortion, Veterinary/microbiology , Actinomycetales Infections/veterinary , Antibodies, Bacterial/blood , Horse Diseases/microbiology , Pregnancy Complications, Infectious/veterinary , Rhodococcus equi/isolation & purification , Aborted Fetus/microbiology , Abortion, Veterinary/pathology , Animals , Feces/microbiology , Female , Horse Diseases/pathology , Horses , Immunohistochemistry/veterinary , Pregnancy , Rhodococcus equi/immunology , Rhodococcus equi/pathogenicity
12.
Aust Vet J ; 85(1-2): 29-31, 2007.
Article in English | MEDLINE | ID: mdl-17300450

ABSTRACT

A 2-year-old intact male domestic shorthaired cat presented with a chronic, nodular, ulcerated, cutaneous lesion on the right thoracic limb. Histological and cytological examination revealed a pyogranulomatous inflammation with basophilic organisms in the macrophages. A virulent form of Rhodococcus equi containing an 87 kb type I (VapA) virulence plasmid was identified from cultures of biopsy samples. This report describes the clinicopathological features, plasmid profile and virulence of this case of R equi infection.


Subject(s)
Actinomycetales Infections/veterinary , Cat Diseases/microbiology , Rhodococcus equi , Actinomycetales Infections/microbiology , Actinomycetales Infections/pathology , Animals , Cat Diseases/pathology , Cats , DNA, Bacterial/analysis , Fatal Outcome , Male , Plasmids/genetics , Rhodococcus equi/genetics , Rhodococcus equi/pathogenicity , Virulence/genetics
13.
Horm Metab Res ; 38(9): 563-9, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16981137

ABSTRACT

We previously showed that tumor necrosis factor-alpha (TNF-alpha) stimulates synthesis of interleukin-6 (IL-6), a potent bone resorptive agent, via p44/p42 mitogen-activated protein (MAP) kinase in osteoblast-like MC3T3-E1 cells. In the present study, we investigated whether phosphatidylinositol 3-kinase (PI3-kinase)/protein kinase B (Akt) is involved in TNF-alpha-stimulated IL-6 synthesis in MC3T3-E1 cells. TNF-alpha induced the phosphorylation of Akt depending upon time. Akt inhibitor, 1L-6-hydroxymethyl-CHIRO-inositol 2-( R)-2- O-methyl-3-O-octadecylcarbonate, significantly suppressed the TNF-alpha-stimulated IL-6 synthesis, but the inhibitory effect was partial. The phosphorylation of Akt induced by TNF-alpha was markedly attenuated by LY294002 and wortmannin, inhibitors of PI3-kinase. Wortmannin and LY294002 significantly reduce the TNF-alpha-induced IL-6 synthesis. On the contrary, the suppressive effects of Akt inhibitor, wortmannin or LY294002 on TNF-alpha-induced phosphorylation of p44/p42 MAP kinase were minor. PD98059, a specific inhibitor of MEK, had little effect on the TNF-alpha-induced phosphorylation of Akt. A combination of Akt inhibitor and PD98059 suppressed the TNF-alpha-induced IL-6 synthesis in an additive manner. These results strongly suggest that PI3-kinase/Akt plays a role in the TNF-alpha-stimulated IL-6 synthesis mainly independent of p44/p42 MAP kinase in osteoblasts.


Subject(s)
Interleukin-6/biosynthesis , Osteoblasts/drug effects , Osteoblasts/enzymology , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Tumor Necrosis Factor-alpha/pharmacology , Androstadienes/pharmacology , Animals , Cells, Cultured , Chromones/pharmacology , Cytokine Receptor gp130/genetics , Cytokine Receptor gp130/metabolism , Flavonoids/pharmacology , Gene Expression Regulation/drug effects , Mice , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Morpholines/pharmacology , Phosphorylation/drug effects , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sirolimus/pharmacology , Wortmannin
14.
Comp Immunol Microbiol Infect Dis ; 29(5-6): 323-33, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17034856

ABSTRACT

In order to assess the Rhodococcus equi infection in three provinces of Turkey (Bursa, Izmir and Istanbul), 696 sera from healthy foals and adult horses were tested by indirect ELISA using a R. equi reference strain (ATCC 6939) as antigen. 103 sera (14.80%) with titres >0.646 resulted positive. Seroprevalence was significantly higher (P=0.0053) in male than in female horses of Istanbul province, although higher antibody titres (mean value) were observed in the female group of Bursa and Izmir provinces with differences estimated between provinces (P=0.0002). Seroprevalence was correlated with age: foals aged less than 1 year (P<10(-4)) and horses from 5 to 10 years old (P=0.018) resulted more infected in Bursa and Izmir provinces. Our findings indicate that R. equi infection actually occurs in all investigated provinces, suggesting the importance of serological survey to diagnose the infection and to prevent the zoonotic risk.


Subject(s)
Actinomycetales Infections/veterinary , Horse Diseases/epidemiology , Rhodococcus equi/immunology , Actinomycetales Infections/epidemiology , Aging , Animals , Antibodies, Bacterial/analysis , Female , Horses , Male , Rhodococcus equi/isolation & purification , Seroepidemiologic Studies , Turkey/epidemiology
15.
Homo ; 57(1): 73-86, 2006.
Article in English | MEDLINE | ID: mdl-16458731

ABSTRACT

Considerable information is available on peak growth velocity characteristics of various body dimensions but the age at minimal velocity (AMV) and the duration of the spurt are not that well documented. Authors applied the mathematical growth model of Preece and Baines (PBGM1) to six longitudinally followed somatometric traits [height, sitting height, iliospinal height (B-ic), upper limb length (a-da), biacromial diameter (a-a), and biiliocristal diameter (ic-ic)] of Japanese boys of Ogi Growth Study. Biological variables derived from the estimated parameters were studied with emphasis on duration and velocity characteristics of the adolescent spurt. Ages for measurements at peak velocities tend to be younger than previously reported non-Japanese ones. Spurt duration in limb measurements was significantly the shortest. Earlier AMV and later age at peak velocity (APV), thus the longest spurt duration, are the characteristic for transverse measurements (a-a, ic-ic). B-ic and a-da had the largest, while a-a and ic-ic had the smallest relative velocity at AMV. Another result for the transverse measurements is that the magnitudes of differences between relative minimal and peak velocities (RMV, RPV) are the largest. It is suggested that a high level of RMV results from early maturation of bones, thus leading to the shortest spurt duration in limb dimensions, while a low level of RMV results from late maturation of the bones, consequently leading to the longest spurt duration in transverse measurements. This tendency of reverse relation was present in the rest of the measurements as well. Transformation of velocity variables (minimal velocity -- MV, peak velocity -- PV) to relative ones, proved to be useful in observing the relation of spurts in measurements.


Subject(s)
Adolescent Development , Adolescent , Body Height , Bone Development , Humans , Japan , Longitudinal Studies , Male , Models, Theoretical , Reference Values
16.
Cancer Res ; 40(2): 477-80, 1980 Feb.
Article in English | MEDLINE | ID: mdl-7356530

ABSTRACT

Peripheral lymphocytes from cancer patients receiving mitomycin C treatment were examined for cytogenetic effects. The treatment consisted of i.v. injections of mitomycin C at a dose of 4 mg given twice a week for 2 weeks. The lymphocytes were cultured in vitro for 72 hr with phytohemagglutinin and 5-bromodeoxyuridine, and then sister chromatid exchanges were scored. Before treatment with mitomycin C, the frequencies of sister chromatid exchanges in lymphocytes of cancer patients were similar to those of healthy controls. After the first and second treatments in vivo with mitomycin C, the frequencies of sister chromatid exchnages increased with time, reached a peak in about 24 hr, and then returned to the pretreatment level in about 48 hr, in contrast to the case of in vitro exposure to mitomycin C. After the third and fourth injections, however, the frequency increased further and did not return to the original level. The significance of the specific kinetics of change in the sister chromatid exchnage frequency after in vivo treatments is discussed in relation to cancer chemotherapy.


Subject(s)
Crossing Over, Genetic/drug effects , Mitomycins/adverse effects , Neoplasms/drug therapy , Sister Chromatid Exchange/drug effects , Adult , Aged , Chromosome Aberrations , Female , Humans , Lymphocytes/drug effects , Lymphocytes/ultrastructure , Male , Middle Aged , Neoplasms/genetics , Time Factors
17.
Cancer Res ; 56(2): 377-83, 1996 Jan 15.
Article in English | MEDLINE | ID: mdl-8542595

ABSTRACT

Zic is a novel zinc finger protein which displays a highly restricted expression pattern in the adult and developing mouse cerebellum and is highly homologous to the recently cloned Drosophila pair-rule gene Opa. To clarify the mechanism for the development of the human cerebellum and its involvement in human nervous system diseases, we have isolated human Zic cDNA and examined its expression by using monoclonal antibody against recombinant Zic protein. The nucleotide sequence of human Zic cDNA is 85% homologous to that of mouse Zic cDNA. Its putative amino acid sequence is highly conserved (> 99%) except for substitution of only two amino acid residues. In situ chromosome hybridization localized the human Zic gene to chromosome band 3q24. Human Zic protein was immunohistochemically detected in the nuclei of the cerebellar granule cell lineage from the progenitor cells of the external germinal layer to the postmigrated cells of the internal granular layer. Furthermore, Zic protein was detected in medulloblastoma (26/29 cases), whereas no other tumors examined (over 70 cases including primitive neuroectodermal tumors) expressed this protein. These findings suggest that Zic is a potential biomarker for medulloblastoma as well as the human cerebellar granule cell lineage.


Subject(s)
Cerebellum/metabolism , DNA-Binding Proteins/biosynthesis , Medulloblastoma/metabolism , Zinc Fingers/physiology , Adolescent , Antibodies, Monoclonal , Brain Neoplasms/diagnosis , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Cerebellum/cytology , Chromosome Mapping , Chromosomes, Human, Pair 3 , Cloning, Molecular , Cytoplasmic Granules/metabolism , DNA, Complementary/genetics , DNA, Neoplasm/genetics , DNA-Binding Proteins/analysis , Diagnosis, Differential , Female , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Medulloblastoma/diagnosis , Medulloblastoma/pathology
18.
Cancer Res ; 51(13): 3544-9, 1991 Jul 01.
Article in English | MEDLINE | ID: mdl-2054791

ABSTRACT

Expression of alpha-amylase genes in thyroid tissues was studied by assaying the total amylase activity as well as by using immunohistochemical and Northern blot analysis. The amylase genes expressed were determined by a combination of the polymerase chain reaction (PCR) and blot analysis using synthetic probes specific for the three known amylase isozyme complementary DNAs. The samples consisted of tissues from 18 human thyroid carcinomas (11 well-differentiated carcinomas, 2 poorly differentiated carcinomas, 1 anaplastic carcinoma, and 4 medullary carcinomas) and 9 specimens of nonmalignant thyroid tissue (2 were from nontumorous regions of resected glands and 7 were thyroid tissue from a patient with Graves' disease). Salivary-type amylase was expressed at a relatively high level in nonmalignant thyroid tissue and well-differentiated carcinoma and could be detected by Northern blot analysis. In poorly differentiated carcinoma, it was detected only by the PCR, while in anaplastic or medullary carcinoma, it was not detected even by the PCR. Thus, the expression of salivary-type amylase was characteristic of well-differentiated follicular cells. These observations suggest that salivary-type amylase expression may be a marker for identifying the histogenesis and stage of differentiation of thyroid cancer. In addition, the AMY2B gene product was detected in all different types of cells examined, although its expression was only detectable by the PCR. Pancreatic type amylase was not detected in any of the samples.


Subject(s)
Isoenzymes/metabolism , Thyroid Gland/enzymology , alpha-Amylases/metabolism , Adult , Aged , Base Sequence , Blotting, Northern , Carcinoma/enzymology , Carcinoma/genetics , Female , Gene Expression , Graves Disease/enzymology , Graves Disease/genetics , Humans , Isoenzymes/genetics , Male , Middle Aged , Molecular Sequence Data , Polymerase Chain Reaction , RNA, Messenger/genetics , Thyroid Neoplasms/enzymology , Thyroid Neoplasms/genetics , alpha-Amylases/genetics
19.
Cancer Res ; 54(14): 3645-50, 1994 Jul 15.
Article in English | MEDLINE | ID: mdl-8033077

ABSTRACT

The ERK gene has been isolated as a genomic DNA encoding a part of the receptor protein-tyrosine kinase which belongs to the EPH subfamily. We previously identified a partial complementary DNA (cDNA) encompassing the catalytic domain of ERK from the expression library of human gastric cancer with an antiphosphotyrosine antibody. Using this cDNA as a probe, the cDNAs encoding mature ERK protein were isolated. The putative mature ERK protein, a total of 967 deduced amino acid residues, showed high homology with chicken Cek5 (92.5%) and mouse Nuk (99.1%). Chromosomal in situ hybridization revealed that human ERK cDNA is localized to chromosome 1p34-35. In Northern blot analysis of normal human tissues, the ERK gene was ubiquitously expressed mainly in cells of epithelial origin but not in the brain. Studies on RNAs from 76 human tumor tissues and cell lines showed that ERK is expressed at higher levels in various tumors of epithelial origin than in corresponding normal tissues, most frequently in gastric cancers (12 of 16, 75.0%). Overexpression of ERK was also detected in one osteosarcoma cell line. These findings suggest that ERK plays some significant role in carcinogenesis in the stomach and other tissues.


Subject(s)
Gene Expression Regulation, Neoplastic , Neoplasms/enzymology , Receptor Protein-Tyrosine Kinases/genetics , Amino Acid Sequence , Base Sequence , DNA, Complementary/isolation & purification , Humans , In Situ Hybridization , Molecular Sequence Data , RNA, Messenger/analysis , Receptor, EphB2
20.
Oncogene ; 12(6): 1259-66, 1996 Mar 21.
Article in English | MEDLINE | ID: mdl-8649828

ABSTRACT

We generated a cDNA expression library from a human neuroepithelioma cell line for detection of novel oncogenes by focus formation assay in NIH3T3 cells. A morphologically unique focus was identified upon transfection and the transforming plasmid was isolated. The transforming gene, designated NET1, encoded a predicted protein species of 54 kDa containing the Dbl-Homology (DH) motif. This motif is also present in other growth regulatory molecules including Bcr, Cdc24, Vav, Ras-Grf, Ect2, Ost, Tim and Tiam1, which have been implicated as regulators of small GTP-binding proteins. NIH3T3 cells transfected with NET1 expression plasmid showed altered growth properties in vitro and were tumorigenic when injected into nude mice. In addition, a 2.5 kb cDNA was isolated from a normal human cDNA library which represented the NET1 proto-oncogene contained a 5' extended open reading frame. The fact that the proto-oncogene failed to induce transformation in NIH3T3 cells suggested that the original NET1 oncogene was activated by 5'-truncation. The 3.0 and 2.4 kilobasepair (kb) transcripts of the NET1 gene was ubiquitously expressed in all tissues examined. Using fluorescence in situ hybridization, we localized the NET1 gene to the short arm of human chromosome 10 at band p15.


Subject(s)
Cell Transformation, Neoplastic/genetics , DNA, Neoplasm/genetics , DNA, Neoplasm/isolation & purification , Neuroectodermal Tumors, Primitive, Peripheral/chemistry , Neuroectodermal Tumors, Primitive, Peripheral/genetics , Oncogene Proteins/genetics , 3T3 Cells/physiology , Amino Acid Sequence , Animals , Base Sequence , Chromosome Mapping , Cloning, Molecular , Exons , GTP-Binding Proteins/genetics , Humans , Mice , Mice, Nude , Molecular Sequence Data , Proto-Oncogene Mas , Sequence Homology, Amino Acid , Transfection , Tumor Cells, Cultured
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