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Japanese spotted fever (JSF) is a tick-borne rickettsiosis caused by Rickettsia japonica. Although the number of JSF cases has been increasing, exceeding 300 per year since 2017, clusters of cases are rare. Here, we report a cluster of seven JSF cases, the first nonfamilial cluster of the disease documented in the Japanese literature, and describe the management of the outbreak through prompt investigation and control-and-prevention measures performed collaboratively by members from the clinical, laboratory, and public health fields. All seven cases in the cluster had visited a cemetery in September or October of 2019. R. japonica was detected in whole-blood and/or skin samples from six patients and in the larvae of Haemaphysalis hystricis collected in a field survey. The evidence suggested that this cluster of cases was caused by the conjunction of two circumstances within a short period of time: an increase in the number of visitors to a cemetery during a Buddhist event and an increase in the number of infectious tick larvae in the cemetery through hatching (vertical transmission from infected females). Delays in the treatment of JSF can lead to severe manifestations. Early interventions through collaborative efforts among members from the clinical, laboratory, and public health fields are important for controlling outbreaks, raising the awareness of the public, and diagnosing and treating patients.

Postpartum thyroiditis (PPT) is characterized by mild thyrotoxicosis occurring within one year of parturition commonly followed by transient hypothyroidism. Having genetic background of autoimmune thyroid disorders is a risk factor for it because the immune reactivation during postpartum period is a trigger for PPT. Pandemic of COVID-19: caused by SARS-CoV-2 infection is a global health problem, and occurrence of Graves' disease and Hashimoto's thyroiditis after the viral infection have been reported but occurrence of PPT with COVID-19 has never been reported. A 29-year-old woman developed general fatigue four and a half months after parturition, and was diagnosed as having PPT: one month before, she had COVID-19. Hereafter, we define the date of delivery as Day 0 to make timeline clear. SARS-CoV-2 infection was diagnosed by PCR on Day 103, its disappearance from the upper airway confirmed on Day 124, and the thyroiditis diagnosed on Day 136. She had been euthyroid on Day 0 and 95, but thyrotoxic on Day 136. Serum thyroglobulin (Tg) concentration was normal in the presence of anti-Tg antibody, other thyroid-related autoantibodies were negative, and by ultrasonography, the thyroid gland was normal in size and no evidence of increased vascularity. Thyroid function returned to normal by Day 172 without any specific drug therapy. In conclusion, although a clear causal relationship could not be found, we documented the world's first case of PPT developed following COVID-19.

Background In June 2013, the fear of adverse events compelled the government to withdraw its recommendation of the Human papillomavirus (HPV) vaccine. Since then, the rate of vaccination among Japanese girls has dropped dramatically. Objective This study aims to assess how the Japanese government's policy change against HPV vaccination influenced the degree to which parents in remote areas in Japan accepted the vaccine for their daughters, and to analyze related factors. Methods A self-administered questionnaire was distributed to parents with daughters attending secondary schools or high schools in two remote areas of the country. Results Out of 700 eligible parents, 246 (35%) responded. The percentage of parents in the “accept group” (those whose daughters had already been vaccinated or those who intended to have their daughters vaccinated) dropped from 54% before the government withdrew its HPV vaccine recommendation to only 4.5% after the withdrawal (1.5% of whom intended to vaccinate and 3% of whom had already had their daughters vaccinated). Vaccine acceptance was higher in parents who were employed as healthcare workers, had been educated through to the completion of high school, had good levels of knowledge about cervical cancer and the HPV vaccine, had low perceived barriers against the HPV vaccine, and received high levels of social support. Conclusion The acceptance of HPV vaccination among parents was low overall. Higher acceptance may be possible if the government restores its recommendation and healthcare providers disseminate appropriate information about the HPV vaccination and cervical cancer.

New Experiments with Spheres-Gas (NEWS-G) is a dark matter direct detection experiment that will operate at SNOLAB (Canada). Similar to other rare-event searches, the materials used in the detector construction are subject to stringent radiopurity requirements. The detector features a 140-cm diameter proportional counter comprising two hemispheres made from commercially sourced 99.99% pure copper. Such copper is widely used in rare-event searches because it is readily available, there are no long-lived Cu radioisotopes, and levels of non-Cu radiocontaminants are generally low. However, measurements performed with a dedicated 210Po alpha counting method using an XIA detector confirmed a problematic concentration of 210Pb in bulk of the copper. To shield the proportional counter's active volume, a low-background electroforming method was adapted to the hemispherical shape to grow a 500-$��$m thick layer of ultra-radiopure copper to the detector's inner surface. In this paper the process is described, which was prototyped at Pacific Northwest National Laboratory (PNNL), USA, and then conducted at full scale in the Laboratoire Souterrain de Modane in France. The radiopurity of the electroplated copper was assessed through Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Measurements of samples from the first (second) hemisphere give 68% confidence upper limits of, 11 pages, 11 figures, 3 tables

We describe the clinical features of a 75-year-old man who presented to our hospital with frequent urination for 3 months. Retrograde urethrography showed extended and continuous narrowing of the stricture in the anterior urethra. Cystourethroscopy showed extended narrowing of the urethral lumen without normal membrane. Urine acid-fast bacillus culture and polymerase chain reaction assays indicated a diagnosis of urethral tuberculosis. Anti-tuberculosis therapy was initiated. Urethral tuberculosis is a very rare cause of urethral stricture in developed countries that can be diagnosed by cystourethroscopy. Urethral tuberculosis should be considered in the differential diagnosis of urethral stricture.

We successfully developed a new photomultiplier tube (PMT) with a three-inch diameter, convex-shaped photocathode, R13111. Its prominent features include good performance and ultra-low radioactivity. The convex-shaped photocathode realized a large photon acceptance and good timing resolution. Low radioactivity was achieved by three factors: (1) the glass material was synthesized using low-radioactive-contamination material; (2) the photocathode was produced with $^{39}$K-enriched potassium; and (3) the purest grade of aluminum material was used for the vacuum seal. As a result each R13111 PMT contains only about 0.4 mBq of $^{226}$Ra, less than 2 mBq of $^{238}$U, 0.3 mBq of $^{228}$Ra, 2 mBq of $^{40}$K and 0.2 mBq of $^{60}$Co. We also examined and resolved the intrinsic leakage of Xe gas into PMTs that was observed in several older models. We thus succeeded in developing a PMT that has low background, large angular acceptance with high collection efficiency, good timing resolution, and long-term stable operation. These features are highly desirable for experiments searching for rare events beyond the standard model, such as dark matter particle interactions and neutrinoless double beta decay events.

We have searched for exotic neutrino-electron interactions that could be produced by a neutrino millicharge, by a neutrino magnetic moment, or by dark photons using solar neutrinos in the XMASS-I liquid xenon detector. We observed no significant signals in 711 days of data. We obtain an upper limit for neutrino millicharge of 5.4×10−12e at 90% confidence level assuming all three species of neutrino have common millicharge. We also set flavor-dependent limits assuming the respective neutrino flavor is the only one carrying a millicharge, 7.3×10−12e for νe, 1.1×10−11e for νμ, and 1.1×10−11e for ντ. These limits are the most stringent yet obtained from direct measurements. We also obtain an upper limit for the neutrino magnetic moment of 1.8×10−10 Bohr magnetons. In addition, we obtain upper limits for the coupling constant of dark photons in the U(1)B−L model of 1.3×10−6 if the dark photon mass is 1×10−3 MeV/c2, and 8.8×10−5 if it is 10 MeV/c2.

We report a case series of 6 patients with confirmed coronavirus disease 2019 (COVID-19) in Wakayama prefecture, Japan. All 6 of the patients tested positive via pharyngeal swab polymerase chain reaction (PCR) tests, and 2 of the 6 were still positive at 3 weeks after onset. All of the patients exhibited bilateral ground glass opacities on computed tomography (CT). This article also reports narrative information on the spectrum of symptoms collected directly from the patients. It would be difficult to triage patients with COVID-19 based on the typical symptoms of fever and/or cough, although PCR and CT are definitive in diagnosis.

To evaluate the diagnostic performance of chest CT to differentiate coronavirus disease 2019 (COVID-19) pneumonia in non-high-epidemic area in Japan. This retrospective study included 21 patients clinically suspected COVID-19 pneumonia and underwent chest CT more than 3 days after the symptom onset: six patients confirmed COVID-19 pneumonia by real-time reverse-transcription polymerase chain reaction (RT-PCR) and 15 patients proved uninfected. Using a Likert scale and its receiver operating characteristic curve analysis, two radiologists (R1/R2) evaluated the diagnostic performance of the five CT criteria: (1) ground glass opacity (GGO)-predominant lesions, (2) GGO- and peripheral-predominant lesions, (3) bilateral GGO-predominant lesions; (4) bilateral GGO- and peripheral-predominant lesions, and (5) bilateral GGO- and peripheral-predominant lesions without nodules, airway abnormalities, pleural effusion, and mediastinal lymphadenopathy. All patients confirmed COVID-19 pneumonia had bilateral GGO- and peripheral-predominant lesions without airway abnormalities, mediastinal lymphadenopathy, and pleural effusion. The five CT criteria showed moderate to excellent diagnostic performance with area under the curves (AUCs) ranging 0.77–0.88 for R1 and 0.78–0.92 for R2. The criterion (e) showed the highest AUC. Chest CT would play a supplemental role to differentiate COVID-19 pneumonia from other respiratory diseases presenting with similar symptoms in a clinical setting.

Gemella is a facultative anaerobic Gram-positive coccus and a rare cause of infective endocarditis (IE). Gram staining may eventually misidentify the organism, which tends to easily decolorize and manifest as either Gram-negative or Gram-variable. Commercial biochemical tests are often used to identify Gemella, but the methods they employ sometimes lack accuracy. A 52-year-old woman was diagnosed with Gemella taiwanensis IE after initial identification of the pathogen as Gemella haemolysans using biochemical tests combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). She was treated successfully with penicillin, gentamicin, and mitral valve replacement. To our knowledge, this is the first case of IE confirmed by 16S rRNA gene and groEL sequencing to have been caused by G. taiwanensis. The accurate diagnosis of rare or difficult-to-identify pathogens is a major challenge for clinical microbiological laboratories. The concurrent use of molecular methods could lead to the recognition of new or different pathogens.

We performed a search for event bursts in the XMASS-I detector associated with 11 gravitational-wave events detected during LIGO/Virgo’s O1 and O2 periods. Simple and loose cuts were applied to the data collected in the full 832 kg xenon volume around the detection time of each gravitational-wave event. The data were divided into four energy regions ranging from keV to MeV. Without assuming any particular burst models, we looked for event bursts in sliding windows with various time width from 0.02 to 10 s. The search was conducted in a time window between − 400 and + 10 , 000 s from each gravitational-wave event. For the binary neutron star merger GW170817, no significant event burst was observed in the XMASS-I detector and we set 90% confidence level upper limits on neutrino fluence for the sum of all the neutrino flavors via coherent elastic neutrino-nucleus scattering. The obtained upper limit was (1.3–2.1) × 10 11 cm − 2 under the assumption of a Fermi-Dirac spectrum with average neutrino energy of 20 MeV. The neutrino fluence limits for mono-energetic neutrinos in the energy range between 14 and 100 MeV were also calculated. Among the other 10 gravitational wave events detected as the binary black hole mergers, a burst candidate with a 3.0 σ significance was found at 1801.95–1803.95 s in the analysis for GW151012. However, no significant deviation from the background in the reconstructed energy and position distributions was found. Considering the additional look-elsewhere effect of analyzing the 11 GW events, the significance of finding such a burst candidate associated with any of them is 2.1 σ .

Double electron capture is a rare nuclear decay process in which two orbital electrons are captured simultaneously in the same nucleus. Measurement of its two-neutrino mode would provide a new reference for the calculation of nuclear matrix elements whereas observation of its neutrinoless mode would demonstrate lepton number violation. A search for two-neutrino double electron capture on $^{124}$Xe is performed using 165.9 days of data collected with the XMASS-I liquid xenon detector. No significant excess above background was observed and we set a lower limit on the half-life as $4.7 \times 10^{21}$ years at 90% confidence level. The obtained limit has ruled out parts of some theoretical expectations. We obtain a lower limit on the $^{126}$Xe two-neutrino double electron capture half-life of $4.3 \times 10^{21}$ years at 90% confidence level as well., 6 pages, 3 figures, accepted for publication in Physics Letters B

Streptococcal toxic shock syndrome (TSS) is a systemic illness usually caused in the setting of infection by group A Streptococcus (GAS). The primary infections are often invasive infections of the respiratory tract or necrotizing infections of the skin and soft tissue, but some infections occur without relevant focus. GAS vaginitis is a rare condition among adult women and is accordingly thought to be uncommon as a cause of streptococcal TSS. Here we report the cases of two postmenopausal women with streptococcal TSS secondary to GAS vaginitis, one aged 55 and one aged 60. Both came to our emergency department with complaints or symptoms of abdominal pain, fever, hypotension, and multi-organ failure. In both cases, the relevant factor associated with streptococcal infection was a recent episode of GAS vaginitis. Both underwent fluid management and 14 days of antibiotic treatment and fully recovered without complications. Vaginitis was likely to be the primary infectious trigger of TSS in these two cases. Intrauterine device insertion, endometrial biopsy, and post-partum state have all been previously reported in TSS patients, and the female genital tract has been described as a portal of entry. GAS vaginitis warrants appropriate treatment as it may progress to severe systemic infection as described.

In June 2013, the fear of adverse events compelled the government to withdraw its recommendation of the Human papillomavirus (HPV) vaccine. Since then, the rate of vaccination among Japanese girls has dropped dramatically.This study aims to assess how the Japanese government's policy change against HPV vaccination influenced the degree to which parents in remote areas in Japan accepted the vaccine for their daughters, and to analyze related factors.A self-administered questionnaire was distributed to parents with daughters attending secondary schools or high schools in two remote areas of the country.Out of 700 eligible parents, 246 (35%) responded. The percentage of parents in the "accept group" (those whose daughters had already been vaccinated or those who intended to have their daughters vaccinated) dropped from 54% before the government withdrew its HPV vaccine recommendation to only 4.5% after the withdrawal (1.5% of whom intended to vaccinate and 3% of whom had already had their daughters vaccinated). Vaccine acceptance was higher in parents who were employed as healthcare workers, had been educated through to the completion of high school, had good levels of knowledge about cervical cancer and the HPV vaccine, had low perceived barriers against the HPV vaccine, and received high levels of social support.The acceptance of HPV vaccination among parents was low overall. Higher acceptance may be possible if the government restores its recommendation and healthcare providers disseminate appropriate information about the HPV vaccination and cervical cancer.

Most mediastinal abscesses result from infections after thoracotomy, esophageal perforation or penetrating chest trauma. This disease is rarely caused by closed blunt chest trauma. All previously reported such cases after closed blunt chest trauma presented with hematoma and sternal osteomyelitis resulting from sternal fracture. Here we report a 15-year-old sumo wrestler who presented with an anterior mediastinal abscess without any mediastinal fracture. The mediastinal abscess resulted from the hematogenous spread of Staphylococcus aureus to a hematoma that might have been caused by a closed blunt chest trauma incurred during sumo wrestling exercises.

A new global digital surface model (DSM) dataset using the archived data of the Panchromatic Remote-sensing Instrument for Stereo Mapping (PRISM) onboard the Advanced Land Observing Satellite (ALOS) has been completed on March 2016 by Japan Aerospace Exploration Agency (JAXA) collaborating with NTT DATA Corp. and Remote Sensing Technology Center, Japan. This project calls “ALOS World 3D” (AW3D), and the dataset consists of the global DSM with 0.15 arcsec. pixel spacing (approx. 5 m mesh) and ortho-rectified PRISM images. JAXA is also processed the global DSM dataset with 1 arcsec. spacing (approx. 30 m mesh) calls “ALOS World 3D 30 m mesh” (AW3D30) based on the AW3D DSM, and released it free of charge on May 2016 as version 1. A weakness of the AW3D30 DSM is remaining void pixels due to cloud covers. This paper describes the AW3D30 partially updates by complementing void pixels using other global digital elevation models, and reports a validation result.

Leptospirosis is not a major disease in urban areas of Japan. We describe a 49-year-old man with leptospirosis, who lived in an urban area and had no history of living in endemic area of leptospirosis. As he worked at a fish market infested with rats, he was suspected of having contracted leptospirosis and received antimicrobial agent treatment. Serum and urinary tests confirmed the diagnosis of leptospirosis. Although it took six days from the onset until treatment initiation, the patient improved in response to receiving ceftriaxone for seven days. Analyzing past reports of Japanese patients with leptospirosis who had no history of overseas travel, we identified 90 patients with courses similar to that of our patient, and the period from onset to treatment initiation was about six days on average (described in 46 cases). Health care providers as well as patients need to recognize that even people with no history of being in an endemic area of leptospirosis may still be at risk of developing this disease depending on occupations and activities.

Erysipelothrix rhusiopathiae is a major causative organism of swine erysipelas, but the infection has rarely occurred in humans. A soft-tissue infection with this organism is described as “erysipeloid.” Most reported cases in human are related to occupational exposure. Endocarditis is a common complication of E. rhusiopathiae bacteremia in human.We report the case of a previously healthy 52-year-old Japanese female who presented with fever and an inflamed hand after being bitten by a stray cat. Blood culture was positive for E. rhusiopathiae, but no sign of infective endocarditis was observed. It may be prudent to consider E. rhusiopathiae infection in patients with animal bites. Keywords: Erysipelothrix rhusiopathiae, Animal bites, Bacteremia

Hemophagocytic syndrome (HPS) can develop as a complication of dengue in rare cases, but its relationship with dengue is not well known. We report a case of dengue-associated HPS with liver involvement and coagulopathy. The patient, a Japanese female traveler who had recently returned from Thailand, had severe complications of dengue infection, but she recovered fully with symptomatic treatment.

The full-length cDNAs of three pepsinogens (Pgs) were cloned from the stomach of newt, Cynops pyrrhogaster, and nucleotide sequences of the full-length cDNAs were determined. Molecular phylogenetic analysis showed that two Pgs, named PgC1 and PgC2, belong to the pepsinogen C group, and one Pg, named PgA, belongs to the pepsinogen A group. The sequences contain an open reading frame (ORF) encoding 385 amino acid residues for PgC1, 383 amino acid residues for PgC2 and 377 amino acid residues for PgA. In addition, all of the three amino acid sequences conserve some unique characteristics such as six cysteine residues and putative active site two aspartic acid residues. All of the pepsinogen mRNAs were detected in the stomach by RT-PCR but not in other organs. Although a slight difference at the time of the start of expression was seen among the three pepsinogen genes, all of them were expressed in the larval stage after hatching. This is the first report on cloning of pepsinogens from urodele stomach.

We treated a case of severe murine typhus in a Japanese traveler after returning from Thailand. Although the disease is typically self-limited or mild, the patient showed shock and multiple organ failure including acute respiratory distress syndrome. Then the patient fully recovered following intensive care and administration of antirickettsial medicines.

We report on a 26-year-old Japanese man who was referred to our hospital because of anal pain and hematochezia. On admission, in addition to his gastrointestinal symptoms, a generalized maculopapular rash was observed, involving the palms of his hands and soles of his feet. His history and physical examination were compatible with syphilis, confirmed by a high syphilis titer on blood examination. Further tests revealed the presence of HIV infection, with a CD4 cell count of 227/microL. Colonoscopy demonstrated a deep ulcer in the lower rectum, although biopsy specimens did not reveal any syphilis spirochetes, or any other specific microorganisms. Intravenous penicillin G was initiated, resulting in a dramatic improvement of the ulcers along with the skin lesions confirming the diagnosis of syphilis proctitis. A rapid plasma reagin titer test performed 3 months after treatment demonstrated significant decrease, indicating successful treatment.

Subjective adverse reactions to metronidazole were analyzed in 111 patients with amebiasis. Metronidazole was administered to 36 patients at a daily dose of 2250 mg and 75 patients at daily doses lower than 2250 mg. The reactions reported included nausea without vomiting in 11 (9.9%) patients, nausea with vomiting in 2 (1.8%), dysgeusia in 2 (1.8%), diarrhea in 1 (0.9%), headache in 1 (0.9%), numbness in 1 (0.9%), dizziness in 1 (0.9%), urticaria in 1 (0.9%), exanthema in 1 (0.9%), and discomfort in 1 (0.9%). Nausea was reported by 28% (10/36) of the patients receiving metronidazole at a daily dose of 2250 mg and 4% (3/75) of the patients receiving lower daily doses. The duration of the metronidazole administration in days was not associated with the appearance of nausea. No life-threatening adverse reactions were identified, and good clinical therapeutic effects were observed in 96% (107/111) of the patients. While metronidazole appears to be a safe anti-protozoal agent for patients with amebiasis, our results indicate that a daily metronidazole dose of 2250 mg is excessive for amebiasis, as it often induces nausea.

The amphibian gastrointestinal tract is remodeled from a larval-type to an adult-type during metamorphosis. In the present study, we examined the products of subtractive hybridization between tadpole and frog stomach cDNAs of Xenopus laevis in order to identify genes expressed specifically in the larval stomach epithelium. A new gene homologous to xP1 was obtained and named xP1-L. In the genome database of Silurana tropicalis, we found a homologue of xP1-L and named it stP1-L. RT-PCR showed that the expression of xP1-L was detected in stage 41/42 tadpoles. In addition, in situ hybridization showed that xP1-L was localized to surface mucous cells of the larval stomach. The H+/K+-ATPase β subunit, a marker gene for manicotto gland cells in the tadpole stomach, was also detected at the same time. However, adult marker genes such as xP1 for surface mucous cells and pepsinogen C (PgC) for oxynticopeptic cells were not expressed in the tadpole stages. The expression of xP1-L gradually decreased towards the metamorphic climax and disappeared after stage 61 when larval-type gastric epithelium is replaced by adult-type. We found that xP1-L was never expressed in surface mucous cells of the adult-type stomach, and xP1, instead of xP1-L, was expressed. During T3-induced metamorphosis, xP1-L expression decreased in the same manner as during natural metamorphosis. Thus, xP1-L is a useful marker for larval surface mucous cells in tadpole stomach. This is the first demonstration of a marker gene specific for the surface mucous cells of the larval stomach.

Six aspartic proteinase precursors, a pro-cathepsin E (ProCatE) and five pepsinogens (Pgs), were purified from the stomach of adult newts (Cynops pyrrhogaster). On sodium dodecylsulfate-polyacrylamide gel electrophoresis, the molecular weights of the Pgs and active enzymes were 37-38 kDa and 31-34 kDa, respectively. The purified ProCatE was a dimer whose subunits were connected by a disulphide bond. cDNA cloning by polymerase chain reaction and subsequent phylogenetic analysis revealed that three of the purified Pgs were classified as PgA and the remaining two were classified as PgBC belonging to C-type Pg. Our results suggest that PgBC is one of the major constituents of acid protease in the urodele stomach. We hypothesize that PgBC is an amphibian-specific Pg that diverged during its evolutional lineage. PgBC was purified and characterized for the first time. The purified urodele pepsin A was completely inhibited by equal molar units of pepstatin A. Conversely, the urodele pepsin BC had low sensitivity to pepstatin A. In acidic condition, the activation rates of newt pepsin A and BC were similar to those of mammalian pepsin A and C1, respectively. Our results suggest that the enzymological characters that distinguish A- and C-type pepsins appear to be conserved in mammals and amphibians.

静岡産業大学国際情報学部では1998年に、学生のメール利用状況とリテラシ教育の関係について調べた。2005年度から、同国際情報学部は情報学部に改組転換し、リテラシ教育の重要性は、以前より高まっているが、一方で、学生を取り巻く状況も大きく変化している。本論文では、学生に新しくアンケート調査を行い、メール・WWWの利用状況と成績の相関を調査した。電子メールに関しては、1998年当時にはっきり見えた傾向が弱まり、かわって「WWWをよく利用する学生の成績がよい」という傾向が強く見られた。

本論文では、「教育の題材(サンプル)」として、また、「教育のための道具(ツール)」として、CGIプログラムを考察する。CGIプログラムは、初級者から上級者にいたるまでの「興味深い題材」になるばかりでなく、初級者が書いたものでさえ、そのまま実用できるという利点を持つことを示したい。また、学生・教員が共同で開発したCGIプログラムを、実際の教育の場で使用することも可能であると考える。

Aminoglycoside-modifying enzymes (AMEs) are major factors that confer aminoglycoside resistance to enterococci. In an epidemiologic study on distribution of 12 AME genes in 534 recent clinical strains isolated from a Japanese hospital, two uncommon AME genes, ant(9)-Ia and a novel aph(2") allele, aph(2")-Ie, were detected. ant(9)-Ia had been reported only in Staphylococcus aureus and encodes spectinomycin adenylyltransferase ANT(9)-I, which confers resistance to spectinomycin. The ant(9)-Ia gene was detected in three strains, a single strain each of Enterococcus faecalis, E. faecium, and E. avium. Nucleotide sequences of ant(9)-Ia from these three enterococcal species were identical to that reported for S. aureus and considered to be located on Tn 554. The new aph(2") allele, designated aph(2")-Ie, was identified in three E. faecium strains. The aph(2")-Ie allele was genetically close to aph(2")-Id reported in E. casseliflavus (93.7% amino acid sequence identity; 96.3% similarity), while distant from aph(2")-Ia, aph(2")-Ib, or aph(2")-Ic (26.3-29.5% amino acid sequence identity). Sequence divergence between APH(2")-Id and APH(2")-Ie was mostly located in amino-terminal half. In contrast, sequences corresponding to the three motifs required for aminoglycoside phosphotransferase were conserved except for a single amino acid. Three E. faecium strains having aph(2")-Ie showed high-level resistance to gentamicin and streptomycin, but not to kanamycin, dibekacin, and tobramycin, unlike enzyme specificity described for aph(2")-Id in E. casseliflavus. Such a difference in resistance phenotype was suggested to be related to amino acid sequence divergence between APH(2")-Id and APH(2")-Ie.

CCAAT/enhancer binding protein delta (C/EBP delta) is one of the transcription factors that have a basic-leucine zipper domain. In mammals, it has been suggested that this transcription factor plays a role in differentiation of adipocytes or in apoptosis of mammary gland epithelial cells. The factor also plays a role in acute-phase response in injury, infection and inflammation. We cloned Xenopus homologues of the C/EBP delta gene from metamorphosing stomach by subtractive hybridization and analyzed spatio-temporal expression pattern of the homologues. Two isoforms of C/EBP delta were isolated and named C/EBP delta-1 and -2. Their deduced amino acid sequences were highly similar to each other (identity, 91.2%). Expression of the C/EBP delta mRNAs in the stomach transiently increased during its metamorphosis-associated remodeling, and the transient up-regulation was also found in thyroid hormone-induced metamorphosis. The C/EBP delta mRNAs were exclusively localized in degenerating larval surface mucous cells, not in newly proliferating and differentiating adult-type epithelial cells. The result suggests a possibility that Xenopus C/EBP delta plays a role in apoptotic cell death of larval-type epithelium during the stomach remodeling.

Through subtractive hybridization, H+/K+-ATPase β subunit mRNA, highly expressed in the larval stomach of Xenopus laevis, was isolated. In situ hybridization demonstrated that the H+/K+-ATPase β subunit mRNA was exclusively expressed in manicotto gland cells of the larval stomach, not in any other cell. Northern blot analysis showed that metamorphosis-associated changes of the H+/K+-ATPase β subunit mRNA expression in the stomach were characterized by high expression in tadpoles, a considerably lower expression in metamorphosing tadpoles, and a re-increase of expression in froglets. Further in situ hybridization showed that the decrease of expression correlated with the degeneration of larval type epithelium in the manicotto gland, while the re-increase correlated with the differentiation of oxynticopeptic cells of the adult type stomach. Moreover, the H+/K+-ATPase β subunit mRNA was expressed in adult epithelial primordia. Such changes were found in thyroid hormone-induced precocious metamorphosis. Based on studies using this ATPase as well as xP1 and PgC (pepsinogen C) as molecular markers, this study discusses a probable cell lineage involved in metamorphosis-associated stomach remodeling. The pH of luminal contents of the larval stomach was found to be lower than 2. In addition, the pH of an isolated stomach changed from 7.2 to lower than 4 after incubation in Ringer's solution, suggesting acid production from the larval stomach. This is the first demonstration of the H+/K+-ATPase-mediated acid production and secretion in the larval stomach of Xenopus laevis. J. Exp. Zool. 301A:992–1002, 2004. © 2004 Wiley-Liss, Inc.