Impact of the COVID-19 pandemic on the incidence of surgical site infection after orthopaedic surgery: an interrupted time series analysis of the nationwide surveillance database in Japan.

BACKGROUND: During the COVID-19 pandemic, hygiene awareness was increased in communities and hospitals. However, there is controversy regarding whether such circumstances affected the incidence of surgical site infections (SSIs) in the orthopaedic surgical field. AIM: To examine the impact of the COVID-19 pandemic on the incidence of SSIs after orthopaedic surgery. METHODS: The medical records of patients having undergone orthopaedic surgery were extracted from the nationwide surveillance database in Japan. The primary outcomes were the monthly incidences of total SSIs, deep or organ/space SSIs, and SSIs due to meticillin-resistant Staphylococcus aureus (MRSA). Interrupted time series analysis was conducted between pre-pandemic (January 2017 to March 2020) and pandemic (April 2020 to June 2021) periods. RESULTS: A total of 309,341 operations were included. Interrupted time series analysis adjusted for seasonality showed no significant changes in the incidence of total SSIs (rate ratio 0.94 and 95% confidence interval 0.98-1.02), deep or organ/space SSIs (0.91, 0.72-1.15), or SSIs due to MRSA (1.07, 0.68-1.68) along with no remarkable slope changes in any parameter (1.00, 0.98-1.02; 1.00, 0.97-1.02; and 0.98, 0.93-1.03, respectively). CONCLUSIONS: Awareness and measures against the COVID-19 pandemic did not markedly influence the incidence of total SSIs, deep or organ/space SSIs, or SSIs due to MRSA following orthopaedic surgery in Japan.

Systematic endoscopic assessment of bleeding sites in severe epistaxis: the role of the S-point and the superior epistaxis.

BACKGROUND: Systematic endoscopic assessment (SEA) of bleeding sites is critical for topodiagnosis and treatment of severe epistaxis, which is not limited to the posterior region. A bleeding site originating from the ethmoidal vasculature, the S-point, has recently been described. The aim of this study is to ascertain the prevalence of each bleeding site in severe epistaxis using a SEA protocol that includes the S-point. METHODOLOGY: Prospective longitudinal study of 51 severe epistaxis patients who underwent 53 SEA under general anesthesia from April 2018 through March 2019. SEA consisted of use of a rigid nasal endoscope; no reduction in blood pressure; no use of topical vasoconstrictor; systematic search of all regions of the nose. Bleeding sites were assigned to either superior or posterior epistaxis. RESULTS: At least one bleeding site was identified in 37 evaluations (69.8%). The S-point was the most common bleeding site (28.3%), followed by the lateral middle turbinate (9.4%), non-S-point upper septum (7.5%), nasal roof (7.5%), and upper lateral wall (7.5%). Superior epistaxis was identified in the most of cases (27 SEA, 50.9%), whereas only 14 SEA (26.4%) identified posterior epistaxis â€" fewer than the 16 SEA that did not identify any bleeding sites (30.2%). There were two recurrences (3.8%). CONCLUSIONS: Systematic endoscopic assessment effectively identified bleeding sites in 69.8% of severe epistaxis. The S-point was the most common bleeding site identified (28.3%). Finally, superior epistaxis corresponded to more than half of the identified bleeding sites, demonstrating the importance of examining this region judiciously in patients with severe epistaxis.

Screen and confirmation of PEG-epoetin ß in equine plasma.

Methods have been developed to screen for and confirm darbepoetin alfa, recombinant human EPO, and methoxy polyethylene glycol-epoetin ß (PEG-epoetin ß) in horse plasma. All three methods screen samples with an enzyme-linked immunosorbent assay (ELISA) and confirm by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This report focuses on PEG-epoetin ß. The ELISA assay was able to detect PEG-epoetin ß at 0.02 ng/mL in 50 µL of horse plasma. Many samples had high background levels of immunoreactivity; however, introducing polyethylene glycol 6000 (PEG 6000) into the samples before the ELISA assay removed the high background and increased the apparent concentrations of PEG-epoetin ß. In samples collected following the administration of 100 µg of PEG-epoetin ß by the intravenous (IV), intramuscular (IM) and subcutaneous (SC) routes, PEG-epoetin ß was detectable up to 72, 144, and 120 h, respectively. The samples were prepared for LC-MS/MS analysis by extraction with anti-rHuEPO-antibodies-coated Dynabeads followed by digestion with trypsin. The LC-MS/MS confirmation method used the multiple reaction monitoring (MRM) scan mode to monitor four precursor-product ion transitions of the EPO-derived peptide T6. All four transitions of T6 were detectable with S/N > 3. The limit of confirmation for PEG-epoetin ß was 1.0 ng/mL in 2 mL of horse plasma. The method successfully confirmed the presence of PEG-epoetin ß in a sample collected from a Mircera®-treated horse. Compared to PEG-epoetin ß, better sensitivity was achieved for darbepoetin alfa and recombinant human EPO. Darbepoetin alfa was detected in horse plasma four days after IM administration of 100 µg.

Biogeochemical features of lipids in endolithic microbial communities in the Ross Desert (McMurdo Dry Valleys), Antarctica.

Endolithic microbial communities inhabiting porous rocks in the cold, dry mountainous regions of Antarctica have been studied extensively as examples of life's adaptations to extreme environments. Here, we examine hydrocarbons and fatty acids occurring in these communities in order to clarify their biogeochemical features with respect to source organisms, microbial activity, fossilization processes and the influence of Gondwanaland sediments. Unusually, long-chain (>C19) n-alkanes and anteiso-alkanes were often the major hydrocarbons in the samples. A suite of n-alkanoic acids (n-C9-n-C32) and long-chain anteiso-alkanoic acids (a-C20-a-C30) were found, along with short-chain iso- and anteiso-alkanoic acids, and n-alkenoic acids. The relationship between long-chain n-alkanoic acids (n-C20-n-C32) and long-chain anteiso-alkanoic acids suggests that these compounds probably originated from the same group of microorganisms, such as bacteria or endolithic lichens, under moderate pH conditions (pH 3-5). Relatively high trans/cis-C16:1 alkenoic acid ratios suggest the presence of unfavorable environmental conditions in the endolithic microbial habitat. Normal-alkenoic/alkanoic acid ratios may be a useful marker for the fossilization of endolithic microbial communities. Thermally matured triterpanes and steranes from fossilized associations on Mount Fleming strongly suggest the presence of Gondwanaland sediments formed during Devonian and Jurassic (400-180 million years ago).

Sporosarcina macmurdoensis sp. nov., from a cyanobacterial mat sample from a pond in the McMurdo Dry Valleys, Antarctica.

Strain CMS 21w(T) was isolated from a cyanobacterial mat sample taken from a pond located in the McMurdo Dry Valleys, Antarctica. Based on its phenotypic, chemotaxonomic and phylogenetic properties, strain CMS 21w(T) was identified as a member of the genus SPOROSARCINA: At the 16S rRNA gene level, CMS 21w(T) exhibited about 93-96 % similarity to all reported species of Sporosarcina and exhibited a maximum similarity of 96 % to both Sporosarcina globispora and Sporosarcina psychrophila. Based on more than 3 % difference at the 16S rRNA gene sequence level and the presence of distinct differences with respect to phenotypic, biochemical and chemotaxonomic features, strain CMS 21w(T) (=MTCC 4670(T)=DSM 15428(T)=CIP 107784(T)) is proposed as the type strain of a novel species of Sporosarcina, Sporosarcina macmurdoensis sp. nov.

Leifsonia rubra sp. nov. and Leifsonia aurea sp. nov., psychrophiles from a pond in Antarctica.

Two unique psychrophilic strains (CMS 76rT and CMS 81yT) were isolated from a cyanobacterial mat sample from a pond in Wright Valley, McMurdo, Antarctica. Both isolates were assigned to the genus Leifsonia, since they were gram-positive, curved rods, non-motile, catalase-positive, contained DL-2,4-diaminobutyric acid, menaquinone MK-11, phosphatidylglycerol and diphosphatidylglycerol, had a high content of anteiso- and iso-branched fatty acids and had a DNA G + C content of 64-66 mol%. In addition, both isolates were related to the five reported species of Leifsonia at a level of about 95-96% 16S rDNA sequence similarity and differed from one another by 2.5%. Strains CMS 76rT and CMS 81yT also differed from one another in many other phenotypic characteristics and exhibited only 30% relatedness at the DNA-DNA level, thus indicating that they represent two different species. Furthermore, these two isolates also showed many distinct differences with respect to the reported species of Leifsonia in terms of their phenotypic characteristics, biochemical properties, chemotaxonomic features, sensitivity to various antibiotics and 16S rDNA similarity, clearly indicating that strains CMS 76rT (= MTCC 4210T = DSM 15304T = CIP 107783T) and CMS 81yT (= MTCC 4657T = DSM 15303T = CIP 107785T) represent the type strains of two novel species of Leifsonia, for which the names Leifsonia rubra sp. nov. and Leifsonia aurea sp. nov. are proposed.

Regression of an unresectable pancreatic tumor following nonmyeloablative allogeneic peripheral-blood stem-cell transplantation.

A 59-year-old female with an unresectable, large pancreatic tumor (10.0 x 8.0 cm(2) on CT scan) underwent nonmyeloablative allogeneic peripheral-blood stem-cell transplantation from her HLA-identical sibling. Pronounced tumor regression and relief from pain without acute graft-versus-host disease (GVHD) were observed following transplantation. The patient is surviving (more than 300 days) after transplantation, with extensive chronic GVHD, and has tumor regression with an 80% reduction in tumor size. The observed clinical course may suggest a graft-versus-tumor effect on the pancreatic tumor following allogeneic stem-cell transplantation.

Contribution of hippocampal place cell activity to learning and formation of goal-directed navigation in rats.

Although extensive behavioral studies have demonstrated that hippocampal lesions impair navigation toward specific places, the role of hippocampal neuronal activity in the development of efficient navigation during place learning remains unknown. The aim of the present study was to investigate how hippocampal neuronal activity changes as rats learn to navigate efficiently to acquire rewards in an open field. Rats were pre-trained in a random reward task where intracranial self-stimulation rewards were provided at random locations. Then, the rats were trained in a novel place task where they were rewarded at two specific locations as they repeatedly shuttled between them. Hippocampal neuronal activity was recorded during the course of learning of the place task. The rats learned reward sites within several sessions, and gradually developed efficient navigation strategies throughout the learning sessions. Some hippocampal neurons gradually changed spatial firing as the learning proceeded, and discharged robustly near the reward sites when efficient navigation was established. Over the learning sessions, the neuronal activity was highly correlated to formation of efficient shuttling trajectories between the reward sites. At the end of the experiment, spatial firing patterns of the hippocampal neurons were re-examined in the random reward task. The specific spatial firing patterns of the neurons were preserved if the rats navigated, as if they expected to find rewards at the previously valid locations. However, those specific spatial firing patterns were not observed in rats pursuing random trajectories. These results suggest that hippocampal neurons have a crucial role in formation of an efficient navigation.

Growth of human squamous cell carcinoma xenografts in mice is inhibited by local angiostatin gene therapy.

The possibility of inhibiting tumor growth by blocking the formation of new tumor vessels has recently received attention. Antiangiogenic tumor therapies have recently attracted intense interest because of their direct endothelial targeting and the absence of drug resistance. Local antiangiogenic gene therapy for cancer offers a potential way to achieve sustained therapeutic release of antiangiogenic substances. As a step toward this goal, we used liposomes complexed to angiostatin cDNA and targeted to human squamous cell carcinoma cell lines in vivo. Tumor cells expressing angiostatin after local gene transfer showed markedly reduced vascularity and contained many apoptotic tumor cells. These results demonstrate the potential utility of liposome-derived angiostatin for adjuvant therapy of oral cancer in humans.

The role of LFA-1 in osteoclast development induced by co-cultures of mouse bone marrow cells and MC3T3-G2/PA6 cells.

Interactions between leukocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) influence the development of osteoclasts. However, little is known about how these adhesion molecules are involved in the process of osteoclast development. This study evaluated the role of LFA-1 and its ligands in osteoclast development and bone resorption. Co-cultures of bone marrow cells from LFA-1-deficient mice and MC3T3-G2/PA6 (PA6) cells were cultured in the presence of 1alpha,25(OH)2D3 and dexamethasone for 7 days. The number of TRAP-positive cells that were generated by bone marrow cells from LFA-1-deficient mice was smaller than that generated by bone marrow cells from wild-type mice. In addition, the bone-resorbing activity of osteoclast-like cells that were generated from LFA-1-deficient mice was lower than that generated by osteoclast-like cells from wild-type mice. Immunofluorescence flow cytometry showed that osteoclast stromal PA6 cells expressed the cell adhesion molecules, ICAM-1 and VCAM-1. When monoclonal antibodies to mice VCAM-1, CD11b or CD18 were added separately to the co-culture system, the number of TRAP-positive cells that were generated from LFA-1-deficient mice was 20-30% smaller than that generated from wild-type mice. The formation of TRAP-positive cells from both LFA-1 deficient and wild-type mice was especially inhibited by anti-CD18 antibody, in comparison to the addition of normal IgG serum. These results suggest that LFA-1 adhesion molecules play a role in osteoclast development by affecting adhesion between stromal cells and osteoclast progenitors before the occurrence of ODF-ODF receptor signaling. CD18 appears to be a key adhesion molecule in cell-to-cell contacts during the early stage of osteoclast development.

Arthrobacter roseus sp. nov., a psychrophilic bacterium isolated from an antarctic cyanobacterial mat sample.

Strain CMS 90rT, a red-pigmented bacterium, was isolated from a cyanobacterial mat sample from a pond located in McMurdo, Antarctica. Based on its chemotaxonomic and phylogenetic properties, strain CMS 90r(T) was identified as a member of group I of Arthrobacter. It shared 16S rDNA similarity of 98% with Arthrobacter oxydans ATCC 14358T and Arthrobacter polychromogenes ATCC 15216T, while DNA-DNA similarities determined for these three organisms were less than 70%. It also differed from all 17 reported Arthrobacter species with A3alpha-variant peptidoglycan in that it possessed a unique peptidoglycan (Lys-Gly-Ala3) and contained galactose, glucose, ribose and rhamnose as cell-wall sugars. These data and the presence of diagnostic phenotypic traits support the description of CMS 90r(T) as a novel species of Arthrobacter, for which the name Arthrobacter roseus sp. nov. is proposed. The type strain is strain CMS 90r(T) (= MTCC 3712T = DSM 14508T).

Planococcus antarcticus and Planococcus psychrophilus spp. nov. isolated from cyanobacterial mat samples collected from ponds in Antarctica.

Thirteen orange-pigmented bacteria associated with cyanobacterial mat samples collected from four different lakes in McMurdo, Antarctica, were isolated. Twelve of the isolates, which were coccoid in shape, were very similar and possessed all the characteristics of the genus Planococcus and represented a new species, which was assigned the name Planococcus antarcticus sp. nov. (CMS 26or(T)). Apart from the phenotypic differences, P. antarcticus differed from all reported species of Planococcus by more than 2.5% at the 16S rRNA gene sequence level. In addition, at the DNA-DNA hybridization level, it exhibited very little similarity either with P. mcmeekinii (30%-35%), P. okeanokoites (26%-29%), or CMS 53or(T) (15%-25%), the three species with which it is closely related at the rRNA gene sequence level (2.5%-2.9%). P. antarcticus also showed only 2.5% difference in its 16S rRNA gene sequence compared with the P. alkanoclasticus sequence. But it was distinctly different from P. alkanoclasticus, which exists only as rods, is mesophilic and phosphatase positive, can hydrolyze starch, cannot utilize succinate, glutamate, or glucose, and cannot acidify glucose. Most important, P. antarcticus and P. alkanoclasticus varied distinctly in their fatty acid composition in that C(15:0), C(15:1), C(16:0), iso-C(16:1), and C(17:0) were present only in P. antarcticus but absent in P. alkanoclasticus. CMS 53or(T), the thirteenth isolate, was also identified as a new species of Planococcus and was assigned the name Planococcus psychrophilus sp. nov. This species was distinctly different from all the reported species, including the new species P. antarcticus, with respect to a number of phenotypic characteristics. At the 16S rRNA gene sequence level, it was closely related to P. okeanokoites (98.1%) and P. mcmeekinii (98%), but with respect to the DNA-DNA hybridization, the similarity was only 35%-36%. The type strain of P. antarcticus is CMS 26or(T) (MTCC 3854; DSM 14505), and that of P. psychrophilus is CMS 530r(T) (MTCC 3812; DSM 14507).

The stress kinase mitogen-activated protein kinase kinase (MKK)7 is a negative regulator of antigen receptor and growth factor receptor-induced proliferation in hematopoietic cells.

The dual specificity kinases mitogen-activated protein kinase (MAPK) kinase (MKK)7 and MKK4 are the only molecules known to directly activate the stress kinases stress-activated protein kinases (SAPKs)/c-Jun N-terminal kinases (JNKs) in response to environmental or mitogenic stimuli. To examine the physiological role of MKK7 in hematopoietic cells, we used a gene targeting strategy to mutate MKK7 in murine T and B cells and non-lymphoid mast cells. Loss of MKK7 in thymocytes and mature B cells results in hyperproliferation in response to growth factor and antigen receptor stimulation and increased thymic cellularity. Mutation of mkk7 in mast cells resulted in hyperproliferation in response to the cytokines interleukin (IL)-3 and stem cell factor (SCF). SAPK/JNK activation was completely abolished in the absence of MKK7, even though expression of MKK4 was strongly upregulated in mkk7(-/-) mast cell lines, and phosphorylation of MKK4 occurred normally in response to multiple stress stimuli. Loss of MKK7 did not affect activation of extracellular signal-regulated kinase (ERK)1/2 or p38 MAPK. mkk7(-/-) mast cells display reduced expression of JunB and the cell cycle inhibitor p16INK4a and upregulation of cyclinD1. Reexpression of p16INK4a in mkk7(-/-) mast cells abrogates the hyperproliferative response. Apoptotic responses to a variety of stimuli were not affected. Thus, MKK7 is an essential and specific regulator of stress-induced SAPK/JNK activation in mast cells and MKK7 negatively regulates growth factor and antigen receptor-driven proliferation in hematopoietic cells. These results indicate that the MKK7-regulated stress signaling pathway can function as negative regulator of cell growth in multiple hematopoietic lineages.

Angiostatin gene therapy inhibits the growth of murine squamous cell carcinoma in vivo.

Tumor growth is an angiogenesis-dependent process and therapeutic strategies aimed at inhibiting angiogenesis are theoretically attractive. Angiostatin has been shown to potently inhibit endothelial proliferation in vitro and tumor growth in vivo. We now show that a shift in the balance of tumor angiogenesis by gene transfer of a cDNA coding for mouse angiostatin into mouse squamous cell carcinoma NRS-1 and SCC-VII cells suppresses tumor growth in vivo. The inhibition of an angiostatin-transfected tumor was accompanied by a marked reduction in vascularity and the presence of many apoptotic tumor cells. However, transfected-angiostatin cDNA does not affect the expression of the vascular endothelial growth factor (VEGF) and VEGF-R2 in the vascular endothelium. The inhibition mechanisms of neovascularization may be mediated independent of VEGF:VEGF-R2 complex. Our data may provide a useful approach for human oral cancer therapy by gene therapy with angiostatin.

Expression pattern of a newt Notch homologue in regenerating newt retina.

We isolated part of a newt Notch homologue, N-Notch, from regenerating newt retina. The spatio-temporal pattern of N-Notch expression was studied by in situ hybridization at different stages of newt retinal regeneration. Proliferating cells were confirmed by the injection of bromodeoxyuridine (BrdU). In the early stage of regeneration, when the retina was one to two cells thick, all proliferating retinal progenitors expressed N-Notch. As the thickness of the retina increased with regeneration, N-Notch expression decreased in BrdU-positive cells on the vitreal side of the retina. Subsequently, presumptive retinal ganglion cells that were BrdU-negative cells appeared at the vitreal edge of the regenerating retina. These differentiating cells did not express N-Notch. Later, N-Notch expression decreased in the BrdU-positive cells on the scleral surface of the retina. Subsequently, presumptive photoreceptor cells that were BrdU-negative cells appeared in this region. These differentiating cells also did not express N-Notch. The proliferating retinal progenitors ceased expressing N-Notch and then stopped dividing during the differentiation of ganglion cells and photoreceptor cells. It was found that retinal regeneration involves the expression of an important developmental signaling molecule, Notch, in retinal progenitors and the expression of Notch ceased as cell differentiation proceeded during retinal regeneration.

Transverse colonic cancer presenting as an anterior abdominal wall abscess: report of a case.

An 81-year-old man who had been aware of a right anterior abdominal mass for 1 week was admitted to our hospital on July 3, 1999, after the mass had perforated and was secreting mucinous purulent material. Computed tomography clearly showed an anterior abdominal wall abscess and a large intraabdominal tumor that contained a fistula-like structure. Barium enema revealed an apple-core sign at the transverse colon, with a fistula that connected the colon to the abscess cavity. Transverse colonic cancer complicated by an anterior abdominal wall abscess was diagnosed, and an extended right hemicolectomy was performed. We did not perform en bloc excision of the full thickness of the anterior abdominal wall, including the abscess, because the defect was determined to be too large to repair. Thus, when curative resection is not feasible, as in our patient, resection of the primary tumor with en bloc partial resection of the adherent parietal wall should be performed if possible, as this procedure has the potential to improve the postoperative quality of life of the patient.

Quantitative TaqMan PCR without a real-time thermal cycler: an assay for fish insulin-like growth factor I messenger RNA.

The reverse transcriptase-polymerase chain reaction, more commonly known as RT-PCR, has become a widely used tool in molecular biology and is now frequently used in monitoring gene expression levels. A number of variations in the RT-PCR technique now exist including TaqMan PCR (5' nuclease assay), which is a useful nonisotopic detection method for the quantification of PCR products. To monitor the formation of these fluorescent amplification products a "real-time" thermal cycler is normally required. In this study, repeated scanning of PCR products in a 96-well plate format showed that a conventional fluorescent plate reader can be used to generate similar results. To demonstrate the power of this approach, the nutritional regulation of insulin-like growth factor I (IGF-I) was investigated in a marine finfish, the snapper (Pagrus auratus). Hepatic IGF-I messenger RNA levels were shown to significantly decrease after 2 weeks of fasting and returned to fed control levels on refeeding. These results demonstrated that a real-time PCR machine was not required to generate this type of quantitative data and that this technology can be adapted for use in most molecular biology laboratories.

Regulation of T cell activation, anxiety, and male aggression by RGS2.

Regulators of G protein signaling (RGS) proteins accelerate the GTPase activity of Galpha protein subunits in vitro, negatively regulating G protein-coupled receptor signaling. The physiological role of mammalian RGS proteins is largely unknown. The RGS family member rgs2 was cloned as an immediate early response gene up-regulated in T lymphocytes after activation. To investigate the role of RGS2 in vivo, we generated rgs2-deficient mice. We show that targeted mutation of rgs2 in mice leads to reduced T cell proliferation and IL-2 production, which translates in an impaired antiviral immunity in vivo. Interestingly, rgs2(-/-) mice also display increased anxiety responses and decreased male aggression in the absence of cognitive or motor deficits. RGS2 also controls synaptic development and basal electrical activity in hippocampal CA1 neurons. Thus, RGS2 plays an important role in T cell activation, synapse development in the hippocampus, and emotive behaviors.

Quantification of optical signals with electrophysiological signals in neural activities of Di-4-ANEPPS stained rat hippocampal slices.

We have quantified the optical signals of synaptically induced neural activities in an in vitro brain slice preparation in terms of electrophysiological signals. The qualification was done using electrophysiologically well known neural activities in the CA1 area of rat hippocampal slices stained with externally applied fluorescent voltage-sensitive dye (VSD; Di-4-ANEPPS). Together with a newly designed CCD-based digital high-speed camera system and epi-fluorescent optics, our improvements were made on a protocol for staining using a newly designed chamber system. These improvements enabled us to make stable and reliable recordings of optical signals and electrophysiological measurements without affecting the physiological status and to make a quantitative comparison between them. The time course and amplitude of the optical signal showed fair agreement with intracellular and extracellular recordings, and was stable over 2 h. The optical signal followed synaptically induced long-term potentiation (LTP) as monitored by the electrophysiological signals. A regional difference in the amount of LTP was found in optical signals and was confirmed in the electrophysiological signals. These results demonstrate the capabilities of our improved method as an alternative but more potent tool to measure the neuronal activities of brain slice in addition to electrophysiological method.