Regulation of thrombocytopoiesis studied by a mathematical model

A mathematical model of thrombocytopoiesis is proposed which accounts for the recent data on its regulation. It is shown that the compensatory response of the system to a decrease in the level of thrombocytes in the blood is controlled by the total amount of thrombocytes and megakaryocytes. The proliferation intensity of megakaryocytes and the total number of thrombocytes reveal, respectively, a lineary and a logarithmical dependence on the total number of thrombocytes and megakaryocytes. The limits of the post-transfusion level of thrombocytes are defined, within which the thrombocytopoiesis is controlled only by the number of thrombocytes. The values of parameters characterizing the behaviour of the thrombocytopoiesis system are calculated.     

Effects of temperature and humidity on the development of eggs of Toxocara canis under laboratory conditions

The influence of temperature and humidity on the survival and development of Toxocara canis eggs in an in vitro model system was investigated. Two soil samples were inoculated with T. canis eggs and maintained at 3% and 50% humidity and temperatures of 19-24 degrees C. Nine soil samples were inoculated with T. canis eggs of which three samples were kept at 4 degrees C with humidities at 3%, 15%, and 30%; three were maintained at 21 degrees C and three more were incubated at 34 degrees C, and at the same three humidity levels. Samples were monitored every 7 days for a total of 2 months, for the presence and development of eggs. With increasing temperature, the number of eggs undergoing development increased (P<0.01); the number of deformed eggs decreased, the number of infective eggs increased (P<0.01), and egg maturation was accelerated. A decrease in the survival of infective eggs occurred at 34 degrees C. An increase in humidity produced a rise in the number of developed eggs at all three temperatures (P<0.01). This study suggests that elevated temperatures accelerated the development as well as the degradation of eggs of T. canis, whereas the range in humidity was directly correlated with egg development.     

A survey of Brucella canis infection in dogs sheltered in Tohoku University School of Medicine (author's transl)]

A total number of 1549 dogs from Miyagi Prefecture and the north coast area of Fukushima Prfecture were surveyed during a year from December 1976 to November 1977 for Brucella canis (B. canis). 173 of 1549 dogs (11.2%) were sero-positive to B. canis, and B. canis was isolated from 55 of 148 dogs examined, giving more than 3.6% for total 1549 dogs. The positivity were higher than that have been reported from other areas. The results of the morphological and biochemical studies for isolates were similar to the reference strain RM6/66, and they were identified as B. canis. No significant difference in sero-positive rate was found between male and female, but isolation rate was higher in female than in male. B. canis was isolated most frequently from the uterus and the spleen of both sexes. The importance of urine and milk as a source of infection was discussed as well as the role of infected focus at non gravid uterus for abortion and fetal infection. A case supposed to be a vertical infection was described also.     

Effects of gangliosides on cell maturation of murine megakaryocytes in a liquid culture system

Formation of platelet-producing megakaryocytes, the cytoplasm of which showed the terminal stage of cell maturation, heavy granulation and platelet-fields delineated with demarcation membranes, was observed in a short-term culture system, using megakaryocyte-enriched bone marrow cell suspension. Approximately 6-8% of the megakaryocytes changed to the platelet-producing megakaryocytes during 12-hour incubation. In the presence of inhibitors of energy metabolism, formation of the platelet-producing megakaryocytes was inhibited, suggesting that the process is dependent on energy producing systems. Ganglioside GD1a increased both the number of total megakaryocytes and the ratio of the platelet-producing megakaryocytes to total megakaryocytes, while GM1 did not influence the number of total megakaryocytes, but increased the ratio. Gangliosides GM2, GM3 and GD1b showed little effect on either the number of total megakaryocytes or the ratio. The results suggest that ganglioside GD1a stimulates at least two steps of megakaryocyte maturation, the change of megakaryocytic progenitors to megakaryocytes and the subsequent maturation of megakaryocytes to the platelet-producing megakaryocytes, while GM1 stimulates only the latter step of the maturation.     

Effect of gangliosides on murine megakaryocytopoiesis in a liquid culture system

Bovine brain gangliosides were applied to primary cultures of murine bone marrow cells to examine the role of gangliosides in development of megakaryocytes. Megakaryocytes in the cultures were detected by staining for a cytoplasmic enzyme, acetylcholinesterase, and divided into two types, 1) immature megakaryocytes which were stained less intensely, and 2) mature ones which were stained intensely. A medium containing total ganglioside fraction from bovine brain increased the number of both immature and mature megakaryocytes in the presence of pokeweed mitogen-stimulated murine spleen cell conditioned medium. Between the two cell types, the number of the mature cells was more significantly increased than the immature cells. The ganglioside GD1a could substitute for the total ganglioside mixture. The results suggested that bovine brain gangliosides potentiated both megakaryocytic proliferation and maturation in vitro.     

The occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs in China

A survey of the occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs was undertaken in the People's Republic of China between October 2008 and October 2009. A total of 600 blood samples were taken from dogs in four cities in China: 300 in Beijing, 150 in Shenzhen, 30 in Shanghai and 120 in Zhengzhou. All samples were tested for the heartworm antigen and antibodies of canine B. burgdorferi, E. canis and A. phagocytophium by using the canine SNAP? 4Dx? test kit. The occurrence of D. immitis, B. burgdorferi, E. canis and A. phagocytophium was 1.17% (7/600), 0.17% (1/600), 2.17% (13/600) and 0.5% (3/600), respectively. In Shenzhen city 2% (3/150), 8.67% (13/150) and 2% (3/150) of samples were positive for D. immitis, E. canis and A. phagocytophium, respectively. The occurrence of heartworm antigen was 0.33% (1/300) in Beijing, 2.00% (3/150) in Shenzhen, 3.33% (1/30) in Shanghai and 1.67% (2/120) in Zhengzhou. We found E. canis and A. phagocytophium only at one site, Shenzhen, while the only occurrence of B. burgdorferi was at Beijing. In conclusion, the dog population in China is at potential risk for D. immitis, B. burgdorferi, E. canis and A. phagocytophium infection, the risk being especially high in southern China.     

Electrocardiographic Examinations in Dogs With Experimental Hepatitis Contagiosa Canis

Dogs with experimentally produced Hepatitis contagiosa canis were subjected to repeated electrocardiographic examinations during the course of the disease. Varying degrees of depression of the STJ and S-T segments and deviations of the T-wave were noted. Serum-electrolyte studies made during the illness showed a decrease of potassium, sodium, and chlorides which did not seem to be the cause of the observed electrocardiographic abnormalities. Circulatory disturbances and changes in the endothelial cells described elsewhere are considered to be the cause of a disturbance in the metabolism and the function of the myocardium with resulting depression of the STJ and S-T segments and deviations of the T-wave.     

Dermatophytes isolated from patients in University Hospital,Kuala Lumpur,Malaysia

A total of 576 dermatophytes were isolated from patients with a variety of skin infections from January 1993 to May 2000. Ten species of dermatophytes were identified: Epidermophyton floccosum (0.7%), Microsporum audouinii (1.1%), M. canis (3.1%), M. gypseum (0.3%), Trichophyton concentricum (3.5%), T. equinum (0.2%), T. mentagrophytes (36.1%), T. rubrum (53.8%), T. verrucosum (0.2) and T. violaceum (1.0%). The body sites most frequently affected by dermatophytes were the buttocks, nails and trunk. Anthropophilic dermatophytes made up 60.1% of the isolates; the most common species was T. rubrum, T. mentagrophytes and M. canis were the two main zoophilic dermatophytes. T. mentagrophytes was isolated from all body sites except the scalp. M. canis was found to be associated with domestic dogs and was not isolated from ethnic Malays. The only geophilic dermatophyte was M. gypseum, an uncommon dermatophyte associated with tinea pedis.     

Effects of hypoxia on megakaryocyte size and number of C3H and BALB/c mice.

In an effort to explain the different platelet production capabilities of both normal and hypoxic male and female C3H and BALB/c mice, megakaryocyte size and number were determined utilizing bone marrow from both normal and hypoxic mice. The results indicate that normal BALB/c female mice have increased numbers of megakaryocytes, but of smaller size compared with either BALB/c male mice or to both sexes of C3H mice. An inverse relationship between the size and number of megakaryocytes was found in both normal and hypoxic mice; therefore, to evaluate total megakaryocyte characteristics, we calculated total megakaryocyte masses (TMM). With hypoxia, megakaryocyte number decreased, whereas megakaryocyte size increased. Despite the increase in megakaryocyte size, hypoxia caused a significant decrease in TMM (P less than 0.005) in all mice, but female C3H mice had higher TMM (P less than 0.05) than did female BALB/c mice. These data show that hypoxia decreases TMM in mice, and that the effect is greater in C3H mice than in BALB/c mice.     

Elutriation for isolation of megakaryocytes

Successful isolation of guinea pig megakaryocytes in large numbers was first achieved with a combination of techniques, taking sequential advantage of the low relative densities and large diameters of most megakaryocytes. Several laboratories have made minor improvements, but this approach retains the disadvantage of losing a significant fraction of the megakaryocyte population, the small immature ones. Counterflow centrifugal elutriation has been shown to eject cells from a chamber progressively, according to their sizes. Because almost all the megakaryocytes are bigger than the other marrow cells, the megakaryocytes can be retained while rejecting the contaminants. With this technology, yields of 1.4-2.0 x 10(6) megakaryocytes from one guinea pig are routine, recoveries have been 93%-94% of the input number of megakaryocytes, and final purities now average 72%. A split-specimen comparison with our previous method found elutriation to provide much greater yield and recovery with at least as great a purification as the density-velocity combination. This new technique was easily adapted to isolation of megakaryocytes in single aspirates from normal human marrow. Fifty-fold purification with near total recovery and a yield of 27,000 megakaryocytes per donor allows easy and reliable cytologic studies. Elutriation appears to be the current method of choice for isolation of megakaryocytes.     

Prevalence and intensity of Toxocara canis (Werner, 1782) in dogs and its potential public health significance in Ile-Ife, Nigeria

A study on the prevalence and intensity of Toxocara canis (Werner, 1782) in dogs was carried out in Ile-Ife, Nigeria. Faecal samples were collected from 269 dogs between January and December 2004, processed by the Kato-Katz technique and then examined for T. canis eggs. The prevalence of T. canis obtained was 33.8%. The intensity of infection, measured as mean egg count per gram of faeces ( +/- SEM) was 393.8 +/- 83.4. The prevalence and intensity of T. canis in dogs aged 0-6 months were significantly higher (P < 0.05) than older age groups. The prevalence and intensity of T. canis infection were significantly higher in males than in female dogs (P < 0.05). Since T. canis is known to cause visceral larva migrans (VLM) in young children, there is the possibility that the high prevalence of T. canis infection obtained in this study might constitute an important risk factor for transmission to humans. Therefore, there is the need to educate the residents of Ile-Ife on the danger of close association of their children with household pets.     

Enzymatic activities of Microsporum canis and Microsporum gypseum strains.

The presence of 11 enzymatic activities, detected by qualitative methods, and 19 enzymes, semi-quantitatively detected by API ZYM system, in strains belonging to Microsporum canis and Microsporum gypseum has been studied. No pronounced differences were noted between Microsporum canis and Microsporum gypseum, although Microsporum gypseum presented in some cases more intense enzymatic activities than Microsporum canis.     

小鼠巨核系祖细胞增殖,分化特性的研究

小鼠骨髓细胞经7d培养后进行细胞形态学观察,可见不同发育阶段的巨核细胞及不同大小的巨核细胞集落。通过计数每个集落中的细胞数,可确定相应祖细胞的有丝分裂能力。结果表明,具有不同有丝分裂能力的祖细胞的体外增殖动力学有所不同。祖细胞的数量与其有丝分裂次数呈负相关(r=-0.986)。进行0、1、2和3次有丝分裂的祖细胞的阿糖胞苷自杀率分别为48.9,58.7,48.0和41.2％;放射敏感性的D_O值(Gy)分别为1.71,1.24,1.03和0.77,D_O值的大小与有丝分裂次数呈负相关(r=-0.958)。经3Gy全身照射后CFU-Meg与CFU-GM的恢复动态过程具有不同特点。     

Babesia Canis Canis, Babesia Canis Vogeli, Babesia Canis Rossi: Differentiation of the Three Subspecies By A Restriction Fragment Length Polymorphism Analysis On Amplified Small Subunit Ribosomal Rna Genes

The parasites Babesia canis and Babesia gibsoni (phylum Apicomplexa) are responsible for canine babesiosis throughout the world. Babesia canis was previously described as a group of three biologically different subspecies, namely B. canis canis, B. canis vogeli, and B. canis rossi. We report partial sequences of small subunit ribosomal RNA gene (ssu-rDNA) of each subspecies amplified in vitro with primers derived from a semi-conserved region of the ssu-rDNA genes in other Babesia species. The polymerase chain reaction combined with a restriction fragment length polymorphism analysis, using HinfI and TaqI restriction enzymes, confirmed the separation of B. canis into three subspecies. These sequences were compared with previously published sequences of other Babesia species. A phylogenetic approach showed that the three subspecies of B. canis belong to the clade of Babesia species sensu stricto where B. canis canis clusters with B. canis rossi whereas B. canis vogeli might form a monophyletic group with the cluster B. divergens and B. odocoilei. Our results show that the three subspecies of B. canis can readily be differentiated at the molecular level and suggest that they might be considered as true species.     

A survey of ectoparasite infestations in stray dogs of Gwang-ju City, Republic of Korea

This study was designed to investigate the incidence of ectoparasite infestation among stray dogs in Gwang-ju City, Republic of Korea. A total of 103 stray dogs collected in the Animal Shelter of Gwang-ju City from November 2003 to August 2005 were investigated in this study. Ectoparasites of one or more genera were detected in 45.6% (47 / 103) of the dogs examined for dermatologic lesions and/or skin scrapings (from 3-5 affected areas). Otodectes cynotis was found to be the most frequent parasite (22.3%, 23 / 103), followed by Sarcoptes scabiei var canis (19.4%, 20 / 103), Ctenocephalides canis (6.8%, 7 / 103), Demodex canis (4.9%, 5 / 103), and Trichodectes canis (1.0%, 1 / 103). Monospecific infestation was found in 83.0% (39 / 47) of the affected dogs, whereas concurrent infestations with 2 or more ectoparasites per animal were found in 17.0% (8 / 47) of the affected dogs. Trichodectes canis is reported for the first time in the Republic of Korea. Dogs less than 1 yr old were more heavily infected than other age groups (66.7%), and small-sized dogs of less than 3 kg body weight were more heavily infected than larger dogs (41.7%).     

Do Toxocara canis larval antigens used in enzyme-linked immunosorbent assay for visceral larva migrans cross-react with AB isohemagglutinins and give false positive results?

A variety of helminth parasites have A and B blood group antigens on their surface. These antigens may cross-react with elevated concentrations of A and B isohemagglutinins in some patients and give false-positive results in the serologic diagnosis of visceral larva migrans caused by T. canis. To clarify this point, serum from patients with visceral larva migrans and elevated T. canis antibody titers as determined by ELISA were absorbed with AB blood cells and retested by ELISA without a demonstrable decline in T. canis antibody titers. Similarly, absorption with T. canis embryonated egg antigens of serum containing elevated levels of anti-A or anti-B isohemagglutinins failed to decrease the isohemagglutinin titer. This indicates that the ELISA using T. canis embryonated egg antigen does not give false positive results with sera containing high concentrations of anti-A or anti-B isohemagglutinins.     

MEGAKARYOCYTOPOIESIS IN MICE

Megakaryocytopoiesis was investigated in young RFM/Unf mice by determining in autoradiograms of marrow smears the labeling index (percentage of cells labeled) of morphologic stages of megakaryocytes at intervals after a single injection of tritiated thymidine. The total maturation time of recognizable megakaryocytes is estimated to be 50–57 hr. The average times spent in the Type I (immature) and Type II (mature) compartments are, respectively, 15 and 42 hr, but we believe that there is a wide variation in these periods among individual megakaryocytes. The frequencies of megakaryocytes of Types I, II and III are 10, 86 and 4%, respectively.     

Molecular Typing Study of the Microsporum Canis Strains Isolated from an Outbreak of Tinea Capitis in School

Tinea capitis is a dermatophyte infection of the scalp that occurs most often in prepubescent children. Tinea capitis may be transmitted by shared use of contaminated hairbrush, by contact with fomites or by direct physical contact with an infected person. Occasionally, outbreak of tinea capitis would happen under some special conditions. Last year, we found an outbreak of tinea capitis in a school due to Microsporum canis. In epidemiological study, we performed the prevalence survey to all of the exposed persons by physical examinations and mycological laboratory tests, including KOH preparation and fungal cultures. We also investigated the environment in the school. In molecular typing study of the M. canis isolated from patients and the environment, random primer amplification polymorphic DNA (RAPD) method, the specific amplification of subrepeat element in the ribosomal DNA nontranscribed spacer (NTS), and the analysis of DNA sequence in the intertranscribed spacer (ITS) of rDNA were performed. The total number of exposed children was seventy-one, among them forty-two were attacked by tinea capitis. The ratio between boy and girl was 13:1. The ages of the patients was ranged from 3.5 years old to 10 years old. Four patients bred cat or dog as pet. Most patients appeared noninflammatory type of tinea capitis and several patients were inflammatory type. Under microscopic examination the invaded hair were all ectothrix. The pathogens isolated from these patients were M. canis. And we also isolated M. canis from the carpet and the pillowcase in the school. The patterns of total strains of M. canis in the RAPD method and PCR amplification of the rDNA NTS region study were identical, and the isolates from patients and the environment contained the same DNA sequences in the ITS region. The outbreak of tinea capitis was caused by M. canis. The M. canis isolated from patients and from the environment were probably the same origin.     

Fecundity and egg output by Toxocara canis in the red fox, Vulpes vulpes

The role of the red fox Vulpes vulpes in the dissemination of eggs of Toxocara canis into the environment is considered with reference to female worm fecundity and egg output in the faeces of infected foxes collected from four localities in southern England. A significant positive correlation was found between female worm size and the number of eggs in the uterus but there was no significant relationship between T. canis worm numbers and egg output in fox faeces. Reliable estimates of worm burdens in foxes could not, therefore, be determined from faecal egg counts alone. The highest mean egg output of 2145.0 epg recorded from adult foxes indicated that fox cubs are not necessarily the main sources of environmental contamination with T. canis eggs. Saturated magnesium sulphate was found to be a more effective flotation solution than zinc sulphate and sodium chloride for recovering eggs from fox faecal samples.     

Capillary form and Function in Dogs With Experimental Hepatitis Contagiosa Canis an Intravital Microvascular Study

Capillary form and function were studied in dogs with experimentally produced Hepatitis contagiosa canis (H.c.c). The observations were made in an intravital microscope (Leitz) and comprised conjuntival and mesenteric vessels as well as connective tissue in the ear chamber. The recordings were made on 16 mm colour film and by single exposures on black and white film. It was found that the flow in the venules was characteristically slowed down with a resulting high arteriolo-venular flow ratio. In the final stages, progressing endothelial swelling occurred impairing corpuscular flow in the capillaries. At the same time, crenated but deformable erythrocytes appeared. In various places cells of small diameter, not identified with certainty, were attached to the endothelial wall. No coating with granulocytes was seen. The thrombocytes were reduced in number and showed no tendency to adhesion. Granular changes were seen in periendothelial granulated cells. Numerous diffuse perivascular haemorrhages occurred.