Prevalence and antimicrobial resistance of Enterococcus spp. isolated from animal feed in Japan.

The rising prevalence of antimicrobial resistance (AMR) of bacteria is a global health problem at the human, animal, and environmental interfaces, which necessitates the "One Health" approach. AMR of bacteria in animal feed are a potential cause of the prevalence in livestock; however, the role remains unclear. To date, there is limited research on AMR of bacteria in animal feed in Japan. In this study, a total of 57 complete feed samples and 275 feed ingredient samples were collected between 2018 and 2020. Enterococcus spp. were present in 82.5% of complete feed (47/57 samples), 76.5% of soybean meal (62/81), 49.6% of fish meal (55/111), 33.3% of poultry meal (22/66), and 47.1% of meat and bone meal (8/17) samples. Of 295 isolates, E. faecium (33.2% of total isolates) was the dominant Enterococcus spp., followed by E. faecalis (14.2%), E. hirae (6.4%), E. durans (2.7%), E. casseliflavus (2.4%), and E. gallinarum (1.0%). Of 134 isolates which were tested for antimicrobial susceptibility, resistance to kanamycin was the highest (26.1%), followed by erythromycin (24.6%), tetracycline (6.0%), lincomycin (2.2%), tylosin (1.5%), gentamicin (0.8%), and ciprofloxacin (0.8%). All Enterococcus spp. exhibited susceptibility to ampicillin, vancomycin, and chloramphenicol. Of 33 erythromycin-resistant isolates, only two showed a high minimum inhibitory concentration value (>128 µg/mL) and possessed ermB. These results revealed that overall resistance to antimicrobials is relatively low; however, animal feed is a source of Enterococcus spp. It is essential to elucidate the causative factors related to the prevalence of AMR in animal feed.

[A framework for assessing essential public health nursing skills and achievement levels required for students graduating from schools that provide education for obtaining a license as a public health nurse in Japan].

PURPOSES: This study aimed to develop a framework for essential skills and the achievement levels necessary for students graduating from schools that provide education for obtaining a license as a public health nurse (PHN) in Japan. METHODS: Two rounds of questionnaire-based investigations using the Delphi methodology were conducted. Subjects were 197 PHNs from municipalities or companies and 146 nurse educators from universities, colleges, junior colleges, or technical nursing schools. RESULTS: (1) The essential skills framework consisted of three (macro, intermediate and micro) levels. Macro-level items were based on the principle of justice, a primary pillar of health care: (A) community assessment to identify health problems; (B) solving and improving particular health problems in collaboration with people to enable them to promote their own health; (C) promoting equitable access and distribution of community resources for health and daily living. Micro-level items had four achievement levels: (I) independent; (II) instructor-guided; (III) laboratory exercise; (IV) theoretical understanding. Micro-level items for A and B had two domains for achievement: individual/family and group/community. (2) In the first round over 70% of respondents said "very important," "important" or "acceptable" for all micro-level items. In the second round, over 90% said all micro-level items fit within macro and intermediate-level items. (3) In the second round, micro-level items attained 70% consensus among PHNs and nurse educators were 71 of 93 (76.3%). Micro-level expression was used for adjustment and the final framework of essential skills yielded 3 macro, 8 intermediate and 59 micro-level items and 95 levels of achievement. (4) In the final framework, the level of achievement for "individual/family" (Macro-level A and B) was almost level I, and for "group/community" almost II or III. The number of micro-level items at level IV for C was 14 of 21 (66.7%). (5) Compared with PHNs, educators generally assigned a more advanced achievement level for the same skill. CONCLUSIONS: This framework offers more clarity to the content and competency of practice and thus should be useful for basic and continuing PHN education. The assumption driving this study was the necessity to improve the nursing educational system, and develop an appropriate curriculum for the PHN national licensure.

Irreparable capsule tears in initial surgery for glenohumeral instability: report of two cases treated with iliotibial band autograft.

Anterior shoulder instability due to massive capsular tearing can usually be repaired by suturing the torn ends together with a satisfactory result. The purposes of this report are to demonstrate the presence of capsular deficiency irreparable by primary sutures even in an initial surgical intervention and to ponder the cause of primary irreparability. We documented the surgical reconstructive technique using the iliotibial band and the subsequent postoperative results in two cases. To our knowledge, there have been no such cases with primarily irreparable capsular tear.

Remodeling of the rat gingiva induced by CO2 laser coagulation mode.

BACKGROUND AND OBJECTIVES: This study was conducted to clarify the morphologic characteristics and subsequent repair process of coagulation necrosis produced by pulsed CO(2) laser irradiation with relatively low fluence, and thereby to evaluate the clinical efficacy of this irradiation mode. STUDY DESIGN/MATERIALS AND METHODS: Wounding of rat gingiva to produce coagulation necrosis was done with a CO(2) laser with a fluence of 326 J/cm(2). The structural characteristics of the wound and subsequent repair process were examined by means of histology, immunohistochemistry, and electron microscopy. RESULTS: At 6 hours after irradiation, the cells in the laser wound appeared histologically intact but had lost the immunoreactivity to antibodies against Hsp47 and exhibited various ultrastructural signs of cell death. This wound area was lined by Hsp70-positive cells. At 1-day post-irradiation, the uptake of BrdU rapidly increased in the adjacent epithelium and connective tissue. The re-epithelization commenced at 1 day and was completed by 7 days. The necrotic tissue gradually became integrated within the newly formed connective tissue and the original contour of the gingiva was retained during the repair process. The repair process of the laser-induced wound progressed more rapidly than that of a scalpel-made wound. CONCLUSIONS: The present study suggests that the coagulation necrosis produced by the low fluence pulsed CO(2) laser does not disturb the repair process but promotes its steady progress and subsequent tissue remodeling. This laser mode will pave the way for more conservative and minimally invasive surgery for treating a wide variety of oral soft tissue disorders.

Finger joint reconstruction with costal osteochondral graft.

Cartilage disorders of the finger joint may influence hand function. Loss of integrity of the finger joint may severely compromise its stability and may lead to degenerative arthritis. Anatomical reduction of the finger cartilage should be as precise as possible in treating cartilage defects due to trauma and other causes. We have repaired cartilage defects in finger joints by arthroplasty with costal osteochondral grafting in 29 patients (30 joints) since 1997. Three patients underwent total joint reconstruction using costal osteochondral grafting for complete bony ankylosis (1 metacarpophalangeal joint and 2 proximal interphalangeal joints). The purpose of this study was to describe the history, indications, and surgical techniques of the costal osteochondral graft for cartilage disorders or defects in the metacarpophalangeal and proximal interphalangeal joints.

Autoregulatory loop and retinoic acid repression regulate pou2/pou5f1 gene expression in the zebrafish embryonic brain.

Zebrafish pou5f1, also known as pou2, encodes a POU-family transcription factor that is transiently expressed in the prospective midbrain and anterior hindbrain during gastrulation, governing brain development. In the present study, we found that the main regulatory elements reside in the proximal upstream DNA sequence from -2.2 to -0.12 kb (the -2.2/-0.1 region). The electrophoretic gel mobility shift assay (EMSA) revealed four functional octamer sequences that can associate with zebrafish Pou2/Pou5f1. The expression of mutated reporter constructs, as well as EMSA, suggested that these four octamer sequences operate in a cooperative manner to drive expression in the mid/hindbrain. We also identified a retinoic acid (RA)-responsive element in this proximal region, which was required to repress transcription in the posterior part of the embryo. These data provide a scheme wherein pou2/pou5f1 expression in zebrafish embryos is regulated by both an autoregulatory loop and repression by RA emanating from the posterior mesoderm.

Altered expression of Smad family members in injured motor neurons of rat.

We examined changes in the expression of Smad family members, which transduce signals from TGF-beta superfamily ligands, following hypoglossal nerve injury. RT-PCR and in situ hybridization revealed that Smad1, 2, 3 and 4 mRNAs were significantly up-regulated in injured side, whereas Smad8 mRNA was down-regulated. Immunohistochemistry and Western blotting analysis confirmed the alterations of Smad1, 2 and 4 in injured neurons. These results suggest that the Smad signaling may be important for nerve regeneration.

Immunohistochemical study of glypican 3 in thyroid cancer.

In 123 patients with thyroid cancer, expression of glypican 3 (GPC3) was immunohistochemically investigated in tissue samples and the biological significance of GPC3 in thyroid cancer was examined. GPC3 was scarcely expressed in the normal thyroid gland, but was dramatically enhanced in certain types of cancers: 100% in follicular carcinoma (20/20 cases) and 70% in papillary carcinoma (48/69 cases). Expression of GPC3 in follicular carcinoma was significantly higher than that of follicular adenoma (p < 0.0019). In contrast, GPC 3 was not expressed in 17 cases of anaplastic carcinoma. A high expression of GPC3 mRNA was confirmed in cancer lesions, which were strongly positive for immunohistochemical staining. In 69 cases of papillary carcinoma, GPC3 was expressed at an early stage, suggesting that GPC3 expression in thyroid cancer is an early event in developing papillary carcinoma. Further studies are required to determine biological functions and molecular mechanisms underlying the upregulation of GPC3 in thyroid cancer.

Fucosylation of N-glycans regulates the secretion of hepatic glycoproteins into bile ducts.

Fucosylated alpha-fetoprotein (AFP) is a highly specific tumor marker for hepatocellular carcinoma (HCC). However, the molecular mechanism by which serum level of fucosylated AFP increases in patients with HCC remains largely unknown. Here, we report that the fucosylation of glycoproteins could be a possible signal for secretion into bile ducts in the liver. We compared oligosaccharide structures on glycoproteins in human bile with those in serum by several types of lectin blot analyses. Enhanced binding of biliary glycoproteins to lectins that recognize a fucose residue was observed over a wide range of molecular weights compared with serum glycoproteins. A structural analysis of oligosaccharides by two-dimensional mapping high performance liquid chromatography and matrix-assisted laser desorption ionization time-of flight mass spectrometry confirmed the increases in the fucosylation of biliary glycoproteins. Purification followed by structural analysis on alpha1-antitrypsin, alpha1-acid glycoprotein and haptoglobin, which are synthesized in the liver, showed higher fucosylation in bile than in serum. To find direct evidence for fucosylation and sorting signal into bile ducts, we used alpha1-6 fucosyltransferase (Fut8)-deficient mice because fucosylation of glycoproteins produced in mouse liver was mainly an alpha1-6 linkage. Interestingly, the levels of alpha1-antitrypsin and alpha1-acid glycoprotein were quite low in bile of Fut8-deficient mice as compared with wild-type mice. An immunohistochemical study showed dramatic changes in the localization of these glycoproteins in the liver of Fut8-deficient mice. Taken together, these results suggest that fucosylation is a possible signal for the secretion of glycoproteins into bile ducts in the liver. A disruption in this system might involve an increase in fucosylated AFP in the serum of patients with HCC.

Fucosylated haptoglobin is a novel marker for pancreatic cancer: a detailed analysis of the oligosaccharide structure and a possible mechanism for fucosylation.

Changes in oligosaccharide structures have been reported in certain types of malignant transformations and, thus, could be used for tumor markers in certain types of cancer. In the case of pancreatic cancer cell lines, a variety of fucosylated proteins are secreted into their conditioned media. To identify fucosylated proteins in the serum of patients with pancreatic cancer, we performed western blot analyses using Aleuria Aurantica Lectin (AAL), which is specific for fucosylated structures. An approximately 40 kD protein was found to be highly fucosylated in pancreatic cancer and an N-terminal analysis revealed that it was the beta chain of haptoglobin. While the appearance of fucosylated haptoglobin has been reported in other diseases such as hepatocellular carcinoma, liver cirrhosis, gastric cancer and colon cancer, the incidence was significantly higher in the case of pancreatic cancer. Fucosylated haptoglobin was observed more frequently at the advanced stage of pancreatic cancer and disappeared after an operation. A mass spectrometry analysis of haptoglobin purified from the serum of patients with pancreatic cancer and the medium from a pancreatic cancer cell line, PSN-1, showed that the alpha 1-3/alpha 1-4/alpha 1-6 fucosylation of haptoglobin was increased in pancreatic cancer. When a hepatoma cell line, Hep3B, was cultured with the conditioned media from pancreatic cancer cells, haptoglobin secretion was dramatically increased. These findings suggest that fucosylated haptoglobin could serve as a novel marker for pancreatic cancer. Two possibilities were considered in terms of the fucosylation of haptoglobin. One is that pancreatic cancer cells, themselves, produce fucosylated haptoglobin; the other is that pancreatic cancer produces a factor, which induces the production of fucosylated haptoglobin in the liver.

Effect of number of fascicle on axonal regeneration in cable grafts.

The effect of number of fascicles on axonal regeneration in cable grafts was examined in the rat cable graft model. The study comprised three experimental groups: the 5f-group, which received 5 fascicles, larger than the host; the 3f-group, in which the total area of the graft fascicles was similar to that of the host; and the 1f-group, which received one fascicle cable graft, smaller in diameter than the host nerve. At the graft segment, well-myelinated fibers were observed both inside and outside the graft fascicles. The three groups showed no difference in morphometric and functional assessment, suggesting that the fibers which regenerated through the outside of the graft might be effectively induced into the distal host. The disproportionate enlargement of the graft fascicle of the 1f-group also increased the fibers passing through it. These findings suggest that a small number of fascicles can induce a larger population of regenerated fibers in the 20-mm cable graft model.

Characterization of BrdU-positive neurons induced by transient global ischemia in adult hippocampus.

Neurogenesis in the brain continues throughout life and is promoted by brain insults including ischemia. There is no critical conclusion, however, about whether proliferated cells acquire neuronal function after ischemia. Transient global ischemia was produced by a four-vessel occlusion procedure in rats (n = 54). To label proliferative cells, rats were administrated with a single dose of 5-bromo-2'-deoxyuridine (BrdU) at 4, 6, 8, 10, 13, or 15 days after ischemia. Increases in BrdU-positive cells were detected in the hippocampal dentate gyrus at 5, 7, and 9 days after ischemia. To determine the phenotype of BrdU-positive cells, BrdU was administrated twice daily for 3 consecutive days during 6 to 8 days after ischemia. A basic helix-loop-helix transcription factor NeuroD at 7 and 14 days and an immature migrating neuronal marker doublecortin at 14 days after ischemia were expressed transiently in proliferative cells. These proliferative cells after ischemia differentiated to the phenotype of neuron at 28 days after ischemia. Furthermore, BrdU-positive neurons showed phosphorylation of extracellular signal-regulated kinase (ERK) by intracerebroventricular injection of N-methyl-D-aspartate (NMDA) at 28 and 56 days after ischemia as seen in surrounding mature neurons. The number of BrdU-positive neurons, which responded to NMDA stimulation, increased with time after ischemia and was greater than that of sham-operated animals. The present study provides evidence for in vivo ERK phosphorylation in response to NMDA stimulation of BrdU-positive neurons in the adult hippocampus after transient forebrain ischemia.

Phosphorylation of extracellular-regulating kinase in NMDA receptor antagonist-induced newly generated neurons in the adult rat dentate gyrus.

Neurogenesis in the adult brain is promoted by various stimulations. NMDA receptor blockade enhances neurogenesis in the hippocampal dentate gyrus. There is no agreed conclusion, however, as to whether newly generated neurons after NMDA receptor blockade obtain functional properties. We investigated the functional maturation of newly generated neurons after NMDA receptor blockade. In the dentate gyrus, 80% of newly generated cells differentiated into the phenotype of mature neurons at 29 days after the single intraperitoneal injection of an NMDA receptor antagonist MK-801. The number of newly generated neurons after MK-801 treatment was significantly greater than that in the saline-treated group. The neurogenic basic helix-loop-helix transcription factor NeuroD protein in the dentate gyrus after MK-801 treatment was expressed transiently in proliferative cells, but not in mature neurons. To determine functional properties of newly generated neurons, we administered NMDA to the lateral ventricle. As an in vivo response, we assessed extracellular-regulating kinase (ERK) phosphorylation. The newly generated neurons showed ERK phosphorylation by NMDA administration as seen in surrounding mature neurons. The number of newly generated neurons, which responded to NMDA receptor stimulation, increased with time after MK-801 treatment. The present study provides evidence that newly generated neurons in the adult hippocampus after NMDA receptor blockade acquire biochemical function in vivo.

Effects of nefiracetam on spatial memory function and acetylcholine and GABA metabolism in microsphere-embolized rats.

The present study aimed to determine whether nefiracetam, N-(2,6-dimethylphenyl)-2-(2-oxo-1-pyrrolidinyl) acetamide, a cognition enhancer, has an effect on learning and memory function in sustained cerebral ischemia, and whether the effect, if any, may accompany modification of the cholinergic or gamma-aminobutyric acid (GABA)ergic system, which are conceived to be involved in the learning and memory function, in the ischemic brain. Sustained cerebral ischemia was induced by the injection of 700 microspheres into the right hemisphere of the rat. The animals were treated once daily with 10 mg/kg nefiracetam p.o. from 15 h after the operation to either 10 days for the water maze study, or 3 or 5 days after the operation for neurochemical examination. Microsphere-embolized rats showed stroke-like symptoms 15 h after the operation and lengthened the escape latency in the water maze task on days 7-10, suggesting a spatial learning dysfunction. The delayed treatment did not reduce the stroke-like symptoms, but effectively shortened the escape latency. The animals at days 3 and 5 after the operation showed decreases in acetylcholine content and choline acetyltransferase activity, which were not prevented by nefiracetam. The microsphere-embolized rats showed decreases in GABA content and glutamic acid decarboxylase activity. The delayed treatment appreciably restored GABA content in the hippocampus on day 5 and reversed glutamic acid decarboxylase activity in both brain regions on day 5. These results suggest that the GABAergic activity rather than the cholinergic activity may be, at least in part, involved in the pharmacological effects of nefiracetam in the ischemic brain.