The influence of celomic membranes and a tunnel between celomic cavities on cancer metastasis in poultry

The roles of celomic membranes and celomic structures in preventing cancer metastasis were examined. The relationship between a tunnel in the peritoneal cavity of hens and the cancer metastasis was also investigated. A tunnel between the right and left dorsal hepatic cavities was found in almost half of healthy hens. The tunnel was also found in 17% of hens with cancer. Neoplastic cells were detected in the ascitic fluid of 79% of hens with effusions. Metastasis was most severe in the intestinal peritoneal and left dorsal hepatic cavities, whereas the right dorsal hepatic cavity showed the least involvement. Severe inflammatory responses were found within the metastatic membranes, resulting in an increase in their thickness. In tumor-bearing hens, crevasse-like splits were present within the membranes at the layers of loose connective tissues. It appears that the splits prevent further invasion by the cancer cells. There were no successful lesions on the inner surfaces of air sacs. It is suggested that the tunnel in the peritoneal cavity enhances cancer metastasis. We also conclude that the celomic membranes and their structure in hens retard the spread of cancer by functioning as a barrier against direct spread of the cancer cells.     

Rapid activation of Mac-1(CD11b/CD18) molecules on macrophages by a new chemotactic factor 'Gasserokine' produced by Lactobacillus gasseri JCM1131T

The chemoattractant activity of a new chemotactic factor, 'Gasserokine' produced by Lactobacillus gasseri JCM1131^T, has been proposed as a novel immunological function of probiotic lactic acid bacteria. The focus of the present study was to understand the mechanism of the chemotaxis induced by Gasserokine, using activation of an adhesion molecule, Mac-1 (CD11b/CD18) on macrophages. The macrophage chemotaxis to Gasserokine was abolished by preincubation of macrophages with the anti-Mac-1 mAb. Gasserokine induced rapid serine phosphorylation of CD18 molecules within 1 min of stimulation, but the effect was short-lived. Substantial tyrosine phosphorylation was observed in CD18-associated protein of macrophages stimulated by Gasserokine. The tyrosine phosphorylation was confirmed in macrophages stimulated with Gasserokine and also serine/threonine phosphorylation was detected on CD18 molecules by laser microscopy using a double immunostaining method. These results suggest that selective activation of intracellular signaling cascades, such as the mitogen-activated protein kinase pathway, are related to the macrophage chemotaxis induced by Gasserokine.     

Comparison of estrogen responsive genes in the mouse uterus, vagina and mammary gland

Female reproductive organs are mainly regulated by estrogen and progesterone. Specifically, the uterus, vagina and mammary gland show organ-specific mitosis and morphological changes during proliferative events, such as estrous cycle, gestation and lactation. The mechanism underlying these organ-specific estrogen-dependent events is still unknown. We examined, therefore, global gene expression in the mature uterus, vagina and mammary gland of ovariectomized adult mice 6 hr after an injection of 5 microg/kg 17beta-estradiol (E2) using a microarray method in order to identify primary E2-responsive genes. Half of the E2 up-regulated genes in the uterus were similar to those in the vagina. E2 up-regulated the expression of Insulin-like growth factor 1 (Igf-1) genes in the uterus and vagina. In the vagina, E2 up-regulated the expression of IGF binding proteins (Igfbp2 and Igfbp5). In the mammary gland, unlike the uterus and vagina, no gene showed altered expression 6 hr after the E2 exposure. These results suggest that expression of Igf-1 and morphogenesis genes is regulated by E2 in an organ-specific manner, and it is supported by the results of BrdU labeling showing E2-induced mitosis in the uterus and vagina except the mammary gland. The differences in organ specificity in response to E2 may be attributed by differences in gene expression regulated by E2 in female reproductive organs. The candidate estrogen-responsive genes in the uterus and vagina identified by profiling provide an important foundation understanding functional mechanisms of estrogen regulating morphogenesis and maintenance of each reproductive organ.     

The effect of low reactive-level laser therapy (LLLT) with helium-neon laser on operative wound healing in a rat model

The effect of low reactive-level laser therapy (LLLT) with a He-Ne laser on operative wound healing was investigated in a rat model. 10-millimeter surgical wounds were created on the backs of Sprague Dawley rats, and animals were assigned to one of eleven groups (n=5). Ten groups received either 8.5 mW or 17.0 mW irradiation of 15 seconds LLLT a day with one of five different irradiation frequencies, i.e. daily (from the 1st to 6th day following surgery), every other day (the 1st, 3rd, and 5th day), on only the 1st day, on only the 3rd day, and on only the 5th day; the 1st day was the day following the surgery. The control group received no irradiation. A skin specimen was harvested from the dorsal thoracic region on the 7th day to measure the rupture strength. The control group had the lowest rupture strength (5.01 N), and the 17.0 mW every other day irradiation group had the highest rupture strength (13.01 N). Statistical differences were demonstrated in the 8.5 mW irradiation setting between the every other day irradiation group and the control group (p<0.05); and in 17.0 mW irradiation setting between the every day irradiation, the every other day, and the 1st day only groups vs. the control group (p<0.01). Histological examination demonstrated that wound healing in the 17.0 mW every other day irradiation group was promoted most significantly such as the prevention of excessive inflammation, increased formation of collagen fibers, and recovery in continuity of tissues. The control group showed poor wound healing and the other experimental groups showed intermediate healing. Thus LLLT with a He-Ne laser was found to promote the healing of operative wounds in the present rat model, in which the most favorable application of LLLT was the 17.0 mW setting of 15 seconds a day with a frequency of every other day.     

Putative host cell receptor for fowl adenovirus detected in gizzard

This work was done to identify a fowl adenovirus (FAV) binding protein in the gizzard, a known target organ for certain strains of FAV serotype 1. By using a virus overlay protein binding assay (VOPBA), a putative FAV binding protein of approximately 200 kDa expressed in the gizzard was detected.     

Effects of Salmonella enterica subsp. enterica serovar Enteritidis vaccination in layer hens subjected to S. Enteritidis challenge and various feed withdrawal regimens

Levels of Salmonella enterica subsp. enterica serovar Enteritidis infection and serum S. Enteritidis antibodies after experimental S. Enteritidis challenge and feed withdrawal were investigated in S. Enteritidis-vaccinated and unvaccinated hens. The results were used to determine whether formalin-inactivated S. Enteritidis vaccination can protect layer hens from S. Enteritidis challenge during feed withdrawal periods. S. Enteritidis infection rates were evaluated from cloacal swabs, eggs and organs. Serum antibody titers to deflagellated S. Enteritidis whole cells (DEWC) and S. Enteritidis FliC-specific 9-kDa polypeptide (SEp 9) were examined by commercial ELISA kits. Cloacal S. Enteritidis recovery rates were lower in the vaccinated than unvaccinated group. Recovery rates of S. Enteritidis from samples increased after feed withdrawal and decreased after re-introduction of feed. S. Enteritidis counts in cloacal swabs were lower in the vaccinated than in the unvaccinated group (P<0.05). More S. Enteritidis-positive eggs were detected from the unvaccinated group. Before S. Enteritidis challenge, the DEWC ELISA titer of the vaccinated group was higher (P<0.05) than the unvaccinated group; subsequently, the S. Enteritidis DEWC ELISA titers of both groups increased gradually. In contrast, only the vaccinated group elicited high SEp-9 antibody titer during post-challenge and feed withdrawal. Additionally, vaccinated hens yielded negative S. Enteritidis isolation rates from egg contents. There is a correlation between negative S. Enteritidis isolation rates and high SEp 9 titers in vaccinated layer hens challenged with S. Enteritidis and subjected to feed withdrawal regimens. These findings suggest the S. Enteritidis vaccination of pullets may protect against S. Enteritidis infection during forced molting and that SEp 9 titer could be a potential indicator of antibody protection against S. Enteritidis infection. The potential of the SEp 9 peptide as an antigen for S. Enteritidis vaccination in the future is worth noting.     

The evaluation of autonomic nervous system activation during learning in rhesus macaques with the analysis of the heart rate variability

The aims of this study were to measure the activity of the autonomic nervous system using heart rate variability (HRV) during learning tasks and to clarify the relationship between learning to overcome a difficult situation and the autonomic nervous system in monkeys. Two young male monkeys (Macaca mulatta) were given simple discrimination learning tasks (DL) and delayed matching to samples tasks (DMTS); Holter-type electrocardiography was done, and HRV was measured. We defined the frequency bands of HRV in rhesus macaques; the low frequency (LF) was 0.01-0.15 Hz, and the high frequency (HF) was 0.15-0.50 Hz. Based on these frequency bands, the LF/HF ratios during learning tasks were analyzed, and a significant increase in the ratio was found during the tasks. The variances in the HF differed between the DL and DMTS tasks; during DMTS tasks, HF variances had a tendency to increase. Our results indicate that increased sympathetic activity accompanied learning and suggest that the parasympathetic nervous system plays a key role during learning, particularly when difficult tasks are being learned.     

Staining patterns for actin and villin distinguish M cells in bovine follicle-associated epithelium

M cells play a central role in the initiation of mucosal immune responses. However, a primary source of difficulty for investigations of this is the lack of an available specific marker for bovine M cells. As M cells possess irregular and short microvilli, we investigated the distribution and localization of the microvillar proteins actin and villin by immunohistochemistry of the gut of calves. In ileum of the calf, actin and villin were clearly and continuously immunostained in the brush border of the villous epithelia, however, discontinuous immunostaining with patches of no staining were observed in follicle-associated epithelium (FAE). Electron microscopy revealed that M cells had irregular microvilli and lacked the typical brush border, and it was inferred that these patches of no staining might be the intercellular crevices of M cells. As the microvilli of M cells were very sparse, there were several areas of weak immunostaining in calf jejunal FAE. These results suggest that M cells in calf FAE are detectable by the absence of staining for actin and villin.     

Multiyear analysis of the dependency of the planting date on rainfall and soil moisture in paddy fields in Cambodia, 2003–2019

The dependencies of the planting date on rainfall and soil moisture in paddy fields in Cambodia were analyzed to quantify farmers’ empirical knowledge regarding their decision of the planting date. Remote sensing data from multiple satellites covering the 2003–2019 period were analyzed. The planting dates in rain-fed paddies ranged from April to August, with large spatial variations and year-to-year fluctuations. In years when planting was suppressed in April and May, planting was extensively enhanced in June and August compared to normal years, and vice versa. Over the northeastern side of Tonle Sap Lake and south of Phnom Penh city, the areas planted in April and May were found to have positive correlations with rainfall and soil moisture, suggesting that wetter-than-average conditions encouraged farmers to plant earlier in the season. In contrast, this relationship was unclear on the western side of Tonle Sap Lake, where the rainfall amounts were larger throughout the year than in other areas in Cambodia. In this region, the relationship between the planting area and soil water availability was either unclear or was even slightly negative from June to August. Since more frequent dry spells have been detected after the onset of the rainy season in recent years, further studies and disseminations of potential changes in dry spells are important for the agronomic adaptation of planting dates under climate change.