Because of demographic change, geriatric patients are becoming a major challenge for traumatology. Multiple trauma patients and patients with proximal femoral fractures are important groups of ...patients in geriatric traumatology. This retrospective study compares two patient groups with different severities of injuries, and analyzes their patient characteristics and short-term outcomes, focusing on functionality upon discharge. The investigation aims to present the characterizing features of both patient groups, and to identify the potential risk factors for early functionality after trauma. The patient collective comprises two patient groups: a polytrauma group with 91 patients, and a femoral fracture group with 132 patients. Under the control of potential influencing factors, the present study showed no significant influence of belonging to either of the patient groups (multiple trauma or proximal femoral fracture) on the mobility status at discharge. Age, known dementia, pre-clinical intubation, and the lowest Hb value were identified as significant influencing factors. Despite their old age and vulnerability, the majority of geriatric patients survive accidents. Further prospective investigations concerning the maintenance or restoration of functionality after an accident are therefore desirable.
Oxidized pyrimidines are mainly repaired by base excision repair, which is initiated by damage-specific DNA glycosylases. NEIL1, the mammalian homolog of Escherichia coli endonuclease VIII and a ...major DNA glycosylase, initiates repair of oxidized pyrimidines. Here, we investigated the expression of two putative variant mouse NEIL1 (mNEIL1) mRNAs―variant 1 ("Neil1 protein" mRNA; BC043297 in the NCBI database) and variant 2 ("unnamed protein" mRNA; AK040802 in the NCBI database)―in normal mouse organs. Reverse transcription-PCR showed that both mRNAs were expressed in total RNA samples from 9 organs. Immunoblot analysis of a nuclear extract from normal mouse liver revealed three bands corresponding to full-length mNEIL1 protein and the two predicted variant proteins. However, neither variant protein, which included an N-terminal enzymatic activity domain deduced from the mRNA variants, were enzymatically active under multiple reaction conditions when expressed as his-tagged recombinant proteins. Nevertheless, recombinant variant 1 protein influenced mNEIL1 activity, while recombinant variant 2 protein had no influence. These results suggest that mNEIL1 mRNA variants are expressed in a variety of organs in normal mice and that variant 1 protein may regulate mNEIL1 activity.
To determine whether Ehrlichia chaffeensis exists in Japan, we used PCR to examine blood from sika deer in Nara, Japan. Of 117 deer, 36 (31%) were infected with E. chaffeensis. The E. chaffeensis 16S ...rRNA base and GroEL amino acid sequences from Japan were most closely related to those of E. chaffeensis Arkansas.
To establish
in situ hybridization and immunohistochemistry based-assays for the detection of porcine cytomegalovirus, routinely processed renal tissue sections from 34 diseased piglets suspected of ...having the infection were obtained and examined. Using hematoxylin and eosin, porcine cytomegalovirus inclusion bodies were found in the nucleus of renal epithelial cells and capillary endothelial cells in the renal medulla in 30 cases. Inclusion bodies corresponding to porcine cytomegalovirus mRNA after
in situ hybridization or porcine cytomegalovirus antigens after immunohistochemistry were easily determined. The cells were characterized by cytomegaly and basophilic intranuclear inclusion bodies. Using
in situ hybridization, porcine cytomegalovirus mRNA were clearly detected in the nucleus and cytoplasm of the cells in 28 of the 30 (93.3%) cases. Using immunohistochemistry, porcine cytomegalovirus antigens were clearly detected in the cytoplasm of the cells in 21 of the 30 (70.0%) cases. Higher specificities and increased intensity of staining was observed with minimal background using
in situ hybridization and immunohistochemistry compared with hematoxylin and eosin. Thus, the two established methods are useful and helpful tools for detecting the presence of a porcine cytomegalovirus infection.
Rhipicephalus appendiculatus serpin-3 (RAS-3), R. appendiculatus serpin-4 (RAS-4) and a 36kDa immuno-dominant protein of R. appendiculatus (RIM36) were reported as candidate antigens for the ...anti-tick vaccine to control ixodid ticks. In the present study, we generated recombinant proteins of RAS-3 (rRAS-3), RAS-4 (rRAS-4) and RIM36 (rRIM36), and assessed their potency as an anti-tick cocktail vaccine in cattle model. RT-PCR analysis showed that RAS-3, RAS-4 and RIM136 transcripts were detected in both adult male and female ticks during feeding. Immunization of cattle with the combination of rRAS-3, rRAS-4 and rRIM36 had raised antibodies against all recombinants and anti-sera had reacted with the molecules from the tick salivary gland extract. Tick infestation challenge demonstrated protective immunity against female ticks, resulting in mortality rates of 39.5 and 12.8 % for the vaccinated and control groups, respectively. Moreover, the mortality rate of Theileria parva-infected female ticks was 48.5 and 10.8 % in the vaccinated and control group, respectively. In order to evaluate the levels of pathogen transmission capacity by T. parva-infected ticks fed on immunized cattle, the occurrence of T. parva in the bovine parotid lymph node and peripheral blood was also determined and quantified by real-time PCR. Although the infection with T. parva could not be protected by the vaccine, the occurrence of pathogen in peripheral blood was delayed 1 to 2 days after the infestation challenge in vaccinated group. These results suggest that this cocktail vaccine plays a role in the prevention of tick infestation.
Although the p16(INK4a) and p21Waf1/Cip1 cyclin-dependent kinase (CDK) inhibitors are known to play key roles in cellular senescence in vitro, their roles in senescence remain rather poorly ...understood in vivo. This situation is partly due to the possibility of compensatory effect(s) between p16INK4a and p21Waf1/Cip1 or to the upregulation of functionally related CDK inhibitors. To directly address the cooperative roles of p16INK4a and p21Waf1/Cip1 in senescence in vivo, we generated a mouse line simply lacking both p16INK4a and p21Waf1/Cip1 genes double-knockout (DKO). Mouse embryonic fibroblasts (MEF) derived from DKO mice displayed no evidence of cellular senescence when cultured serially in vitro. Moreover, DKO MEFs readily escaped Ras-induced senescence and overrode contact inhibition in culture. This was not the case in MEFs lacking either p16INK4a or p21Waf1/Cip1, indicating that p16(INK4a) and p21Waf1/Cip1 play cooperative roles in cellular senescence and contact inhibition in vitro. Notably, we found the DKO mice to be extremely susceptible to 7,12-dimethylbenz(a)anthracene/12-O-tetradecanoylphorbol-13-acetate-induced skin carcinogenesis that involves oncogenic mutation of the H-ras gene. Mechanistic investigations suggested that the high incidence of cancer in DKO mice likely reflected a cooperative effect of increased benign skin tumor formation caused by p21Waf1/Cip1 loss, with increased malignant conversion of benign skin tumors caused by p16(INK4a) loss. Our findings establish an intrinsic cooperation between p16INK4a and p21Waf1/Cip1 in the onset of cellular senescence and tumor suppression in vivo.
Ehrlichia canis is an obligate intracellular bacterium that infects the macrophage–monocyte cells of dogs, causing canine monocytic ehrlichiosis. Interferon-γ (IFN-γ), along with other cytokines, ...mediates the immune response to such intracellular bacterial invasions. To determine the role of IFN-γ in the immunity of dogs to E. canis infection, peripheral blood mononuclear cells (PBMC) and white blood cells (WBC) were collected from E. canis-infected dogs and added to a culture of E. canis in DH82 cells. The number of E. canis inclusion-positive cells was significantly reduced in cultures containing PBMC and WBC from E. canis-infected dogs compared to uninfected dogs. However, this resistance was inhibited by the addition of an anti-dog IFN-γ antibody. Resistance was also observed when PBMC were added to the Cell Culture Inserts, which prohibited contact of PBMC to DH82 cells, while allowed the diffusion of soluble cell products. The results of this study indicate that resistance was not dependent on cell to cell contact, but was associated with soluble cell products, such as IFN-γ. The addition of recombinant canine IFN-γ to the E. canis culture also reduced the number of infected cells. A commercial recombinant canine IFN-γ, which is sold in Japan, was also effective at reducing E. canis-infected cell number. These results indicate that IFN-γ has an inhibitory effect on the frequency of E. canis-infected cells in vitro and that contact between effector and target cells is not necessary for the resistance.
The intercalation reaction of diclofenac sodium (DFS) with layered inorganic compounds, γ-titanium phosphate (γ-TiP), proton type titanium oxide (H-TiO2) and sodium type synthetic mica (Na-TSM), was ...examined on. The direct reaction of DFS in ethanol–water mixed solvent resulted in the large amount accommodation of DFS. The amount of intercalated DFS was the order of γ-TiP>H-TiO2>Na-TSM corresponding to the order of acidity. The intercalation using phospholiopids was also examined to assist the intercalation reaction. However, the amount of intercalated DFS was rather small in comparison with those in the direct reaction. DFS accommodated in γ-TiP dissolved into neutral and basic buffer solution stoichiometry while scarcely dissolved in the acidic solution. The mechanism of the intercalation and reverse dissolution was successfully accounted according to the ion-exchange mechanism between Na+ in DFS and H+ in γ-TiP. The dissolution from tablet of DFS/γ-TiP intercalation compound was examined by using a disintegrator. It was found that the dissolution rate appropriately controlled by mixing the disintegrator. The present results suggested the different possibilities in the clinical field to use layered inorganic compounds such as drug delivery system (DDS).
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