Characteristics of doctors' fatality due to COVID-19 in Western Europe and Asia-Pacific countries.

Under the coronavirus disease 2019 (COVID-19) pandemic, the deaths of healthcare professionals have been increasingly reported worldwide. We performed a cross-sectional, observational study using news reports on the websites among selected countries as of April 2020. We found 120 dead medical doctors due to COVID-19 in Western Europe and Asia-Pacific countries; 67 in Italy (47 in the Northern part), 34 in China (22 in Hubei), 6 in France, 4 in the UK, the USA and Spain and 1 in South Korea, respectively. Among them, 90% were men, and specialties were reported as general practitioners for 30% and as physicians for 11.6%. The overall proportions of dead medical doctors amounted to 1.9 per 10 000 confirmed cases and 30.2 per 10 000 dead cases, respectively. Proactive measures are warranted to protect doctors especially who often encounters with COVID-19 patients.

Understanding the Environmental Impact of a Mine Dam Rupture in Brazil: Prospects for Remediation.

The rupture of the Fundão mine dam in Mariana municipality, Minas Gerais State, Brazil, spilled the tailings across the Doce River basin. These tailings, composed of residues discarded from the beneficiation of iron ore, are rich in SiO and AlO, as well as some ether amine compounds and NaOH. The aim of this study was to assess the distribution of these sediments, as well as their effect on the riparian zones reached, as compared with preserved sites. Sediment deposition in the river resulted in a morphological change from a meandering profile to a braided aspect. The nutrient and mineral content (P, K, Ca, Mg, Cu, Fe, Mn, Zn, and NO) and soil organic matter of the sediments were depleted, whereas NH, Na, and pH increased. A random presence of ether amines in the sediments was confirmed by quantitative and chromatographic analyses, with concentrations ranging from 0 to 57.8 mg kg; Na reached values as high as 150 mg kg. The impact of the dam tailings on biota was assessed by estimating total microbial biomass (phospholipid fatty acids), which were depleted in sediments relative to soils from preserved sites. Overall plant mortality, as well as a low resilience capacity, were also observed. Ether amines and Na present in the sediments had a strong toxic effect in the environment. Identification of these substances as the main impact factors will help guide future remediation efforts.

Thymic hyperplasia in patients with Graves' disease. Identification of thyrotropin receptors in human thymus.

Thymic size and density were studied in 23 untreated patients with Graves' disease and 38 control subjects using computed tomography. Both thymic size and density were higher in untreated patients with Graves' disease than in control subjects in the age-matched group. After treatment with antithyroid drugs, both thymic size and density were significantly reduced, with a concomitant decrease in thyrotropin receptor antibodies. PCR of human thymic cDNA using primers for human thyrotropin receptor amplified a fragment in a size expected for the receptor, and its nucleotide sequence was identical to human thyrotropin receptor cDNA in the thyroid. Northern blot analysis of human thymic poly(A)+ RNA demonstrated the presence of the full length form of thyrotropin receptor mRNA. Western blot analysis of human thymic membrane using anti-thyrotropin receptor peptide antibodies demonstrated a band of 100 kD that was also observed in the thyroid membrane. Immunohistochemistry of thymic tissue using mouse antihuman thyrotropin receptor monoclonal antibodies demonstrated the immunostaining of epithelial cells. These results indicate that thymic hyperplasia is apparently associated with Graves' disease and suggest that thymic thyrotropin receptor may act as an autoantigen that may be involved in the pathophysiology of development of Graves' disease.

Protection of hexaprenyl-diphosphate synthase of Micrococcus luteus B-P 26 against inactivation by sulphydryl reagents and arginine-specific reagents.

Hexaprenyl-diphosphate synthase from Micrococcus luteus B-P 26 has been shown to comprise two essential components, designated as components A and B. Treatment of the synthase with sulphydryl reagents (N-ethylmaleimide, iodoacetamide or p-chloromercuribenzoate) or arginine-specific reagents (2,3-butanedione, 1,2-cyclohexanedione or phenylglyoxal) resulted in a rapid loss of the component B activity. In contrast, component A was resistant to treatment with such reagents, retaining the initial activity almost completely. Farnesyl diphosphate, isopentenyl diphosphate, farnesyl monophosphate and inorganic pyrophosphate protected the synthase against the inactivation by N-ethylmaleimide, farnesyl diphosphate being the most effective. The presence of Mg2+ was essential for the protection by isopentenyl diphosphate and inorganic pyrophosphate. For protection of the synthase activity against the inactivation by 2,3-butanedione, the presence of farnesyl diphosphate, isopentenyl diphosphate and Mg2+ was more effective than that of the individual substrates and Mg2+. Inorganic pyrophosphate provided substantial protection. In the absence of component A, the component B activity was not protected by any substrates or its analogue. These results suggest that the catalytic site of the synthase is formed by cooperative interaction between components A and B, and that cysteine and arginine residues on component B play important roles in the synthase activity.

Primary amyloidosis with familial vitreous opacities: an unusual case and family.

Peripheral neuropathy was not found even six to ten years after the onset of visual symptoms in a family with primary amyloidosis, except in the propositus at the terminal stage. The propositus had mainly ocular and CNS involvement. An ocular manifestation, the vitreous opacity, was the only involvement in the family members, in spite of the long clinical course. This family may have a different type of familial primary amyloidosis from that previously reported.

Preparation of monoclonal antibodies against brain natriuretic peptide and their application to radioimmunoassay and passive immunization.

Two monoclonal antibodies (mAbs) directed toward brain natriuretic peptide (BNP), KY-BNP-I and KY-BNP-II, have been produced. Both mAbs against BNP possessed high affinities for BNP, with association constants (Ka) of 4.0 X 10(9) M-1 (KY-BNP-I) and 2.0 X 10(10) M-1 (KY-BNP-II). With these mAbs, specific RIAs for BNP have been established. The least detectable quantities of BNP were 5 pg/tube (KY-BNP-I) and 1 pg/tube (KY-BNP-II). Cross-reactivities of alpha-human and rat atrial natriuretic polypeptides were less than 0.001%. These RIAs detected BNP-like immunoreactivity (BNP-LI) not only in the porcine brain but also in the canine brain, with the highest concentration in the medulla oblongata. These RIAs also detected BNP-LI in both the porcine and canine hearts and in the porcine plasma. The iv pretreatment of purified mAb[KY-BNP-II] almost completely blocked the hypotensive action of iv administered BNP in rats, with the concomitant suppression of BNP-induced increase of the plasma cyclic GMP level. These results indicate that our mAbs against BNP will serve as a useful tool for the elucidation of the physiological and pathophysiological significance of BNP as a neuropeptide and as a hormone.

Effect of CpG methylation on expression of the mouse imprinted gene Mest.

We previously reported isolation of the mouse gene, Mest (mesoderm-specific transcripts), which is mapped to the proximal part of chromosome 6 and predominantly expressed in the mesoderm and its derivatives during development. Peg1, a paternally expressed gene isolated by a systematic screening of imprinted genes, was recently demonstrated to be identical to Mest. We and others have shown that the human homolog (MEST) of Mest is also imprinted so as to be expressed from the paternal copy and maps to 7q32. To study transcriptional regulation of Mest/Peg1, we examined the effect of DNA methylation on its expression. In the embryonal carcinoma (EC) cell line, MC12, from which Mest was originally isolated, the 5'-region harboring presumptive promoter of the gene was undermethylated. On the other hand, C4XX, a subclone of MC12 which had lost expression of Mest, was characterized by extremely high levels of methylation in the 5'-region, demethylation of which resulted in activation of Mest. Furthermore, a methylated reporter construct with the luciferase gene under the control of the putative promoter region of Mest was not competent to produce luciferase activity in MC12 cells. These results suggest a suppressive role for DNA methylation in Mest expression. However, neither methylated nor unmethylated reporter constructs showed luciferase activity in a primary culture from the adult kidney, in which Mest is down-regulated despite apparent unmethylation of the paternal allele. Taken together, the data suggest that there are probably two modes of regulation for the Mest gene; one being a methylation-dependent mechanism that regulates imprinted expression of Mest during development, and the other being a methylation-independent mechanism that is involved in down-regulation of Mest in adult tissues.

Participation of type I interferon in the decreased virulence of the UL13 gene-deleted mutant of herpes simplex virus type 1.

We isolated a UL13 gene-deleted mutant of the herpes simplex virus type 1 (HSV-1) strain VR3 (VRDelta13) and its revertant virus (VRDelta13R). This deletion mutant still had virus host shutoff (vhs) activity, although a previous report had suggested the possibility of a functional relation between the UL13 product, that is protein kinase (PK), and vhs activity. We compared the in vivo growth of these viruses in BALB/c mice. VRDelta13 was cleared in the early period of intraperitoneal infection. VRDelta13 had a higher sensitivity to the mouse type I interferon (IFN) and showed a higher level of IFN induction in the study period of infection than did VR3 and VRDelta13R. These results suggest that a nonspecific antiviral response (i.e., the IFN system) may contribute to this rapid inhibition of viral replication in vivo.

Spontaneous reactivation of the inactive X chromosome in mouse embryonal carcinoma cells.

The mouse embryonal carcinoma cell line MC12 carries two X chromosomes, one of which replicates late in S phase and shares properties with the normal inactive X chromosome and, therefore, is considered to be inactivated. Since the hypoxanthine phosphoribosyl transferase (HPRT) gene on the active X chromosome is mutated (HPRT(NDASH;)), MC12 cells lack HPRT activity. After subjecting MC12 cells to selection in HAT medium, however, a number of HAT-resistant clones (HAT(R)) appeared. The high frequency of HAT resistance (3.18 x 10(-4)) suggested reactivation of HPRT(PLUS;) on the inactive X chromosome rather than reversion of HPRT(NDASH;). Consistent with this view, cytological analyses showed that the reactivation occurred over the length of the inactive X chromosome in 11 of 20 HAT(R) clones isolated. The remaining nine clones retained a normal heterochromatic inactive X chromosome. The spontaneous reactivation rate of the HPRT(PLUS;) on the inactive X chromosome was relatively high (1.34 x 10(-6)) and comparable to that observed for XIST-deleted somatic cells (Csankovszki et al., 2001), suggesting that the inactivated state is poorly maintained in MC12 cells.

Primary lung cancer SPECT imaging with pentavalent technetium-99m-DMSA.

UNLABELLED: To assess the clinical role of 99mTc(V)-DMSA in primary lung cancers, SPECT imaging was performed on 31 patients with suspected lung cancer. METHODS: Planar and SPECT images were obtained at 3 to 4 hr after intravenous injection of approximately 555 MBq 99mTc(V)-DMSA. Two uptake ratios (the maximum counts/pixel in the lesion to the average counts in normal tissue) were calculated. RESULTS: Various types of primary lung cancers (adenocarcinoma, squamous-cell carcinoma, small-cell carcinoma, large-cell carcinoma and bronchial carcinoid tumor) were imaged by 99mTc(V)-DMSA SPECT. Approximately 90% of the lung carcinomas showed increased uptake and were clearly demonstrated by SPECT images. Four cases incidentally revealed osseous metastatic lesion. Three benign lesions did not show increased uptake. Three cases were false-negative and there were no false-positive cases for the primary lesions. CONCLUSION: Technetium-99m(V)-DMSA SPECT images demonstrated approximately 90% of the primary lung cancers. Uptake ratios were higher in squamous-cell carcinomas than adenocarcinomas. Evaluation of mediastinal tumor extension and nodal metastatic lesion was very difficult by high blood-pool activity in the major cardiovascular structures due to slow blood-pool clearance. However, 99mTc(V)-DMSA SPECT imaging was very useful for detecting primary lung cancers and metastatic lesions to the osseous structures.

Gender-dependent effect of L-NAME on polycystic kidney disease in Han:SPRD rats.

To determine whether the renal nitric oxide (NO) system has a role in the pathogenesis of polycystic kidney disease (PKD) in Han:Sprague-Dawley (SPRD) rats, the NO synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), 70 mg/L, or L-arginine, 0.5 g/L, was administered to heterozygous diseased (cy/+) and homozygous normal animals. Urine nitrate and nitrite excretion was reduced by L-NAME treatment and, in the male cy/+ rats, increased by L-arginine administration. The administration of L-NAME significantly increased blood pressure in all groups, whereas L-arginine had no effect. L-NAME and L-arginine had a modest but significant overall effect on the severity of cystic disease in male rats, reflected by relative kidney weights and cyst volume densities. This effect was gender dependent because it was not observed in female animals. The administration of L-NAME resulted in a significant increase in plasma creatinine concentration of the cy/+ rats, which was more marked in male than female animals. These observations support the recently reported gender differences in the renal NO system and a small role for NO synthesis that can be inhibited by L-NAME in the pathogenesis of PKD in Han:SPRD rats. These observations do not exclude a more important role for the endogenous renal NO production in the pathogenesis of PKD in view of a recent report of a major NOS resistant to conventional inhibitors in the rat kidney.

Synthesis and anti-influenza virus activity of novel pyrimidine derivatives.

Efficient synthetic routes of 2-amino-4-(omega-hydroxyalkylamino)pyrimidine derivatives were investigated in relation to the anti-influenza virus activity of these compounds. The derivatives in which cyclobutyl and cyclopentyl groups were introduced to the beta-position of the aminoalkyl group (especially the cyclobutyl group substituted by a phenylalkyl group at the 3'-position) resulted in improved antiviral potency: i.e. an average 50% effective concentration for inhibition of plaque formation (EC50, microM) of 0.1-0.01 microM for both types A and B influenza virus. The antiviral efficacies were in the order of amino group > hydroxyiminomethyl group > halogen substitution at the 5-position, and chlorine or methoxy group > hydrogen at the 6-position of the pyrimidine ring. The antiviral indices of these compounds were 2-6 with respect to the 50% inhibitory concentration for cell proliferation (IC50, microM) for growing cells, but > 500 to > 10(4) with respect to the IC50 for stationary cells, indicating that these compounds may be efficacious for the topical treatment of influenza virus infection.

Undecaprenyl diphosphate synthase from Micrococcus luteus B-P 26: essential factors for the enzymatic activity.

An undecaprenyl diphosphate synthase fraction, which was free of other prenyltransferases and was active without the addition of detergent or phospholipid, was obtained by Sephadex G-100 chromatography of cell-free extracts of Micrococcus luteus B-P 26 cells. The addition of small amounts of Triton X-100 to this fraction caused a marked loss of the enzyme activity, but the activity was gradually restored as further detergent was added. When the enzyme fraction was chromatographed on DEAE-cellulose, the synthase was partially purified, but the activity was not detected unless assayed with addition of the detergent or a lipid fraction of this bacterium. Among the three phospholipids isolated from this bacterium, cardiolipin and phosphatidylglycerol had a marked effect in activating lipid-depleted undecaprenyl diphosphate synthase, but O-lysylphosphatidylglycerol, which occurs prominently in this bacterium, had little effect.

Highly specific enzyme immunoassay for human chorionic gonadotropin.

A highly specific enzyme immunoassay for determining hCG was established by using beta-D-galactosidase as label. In order to increase the specificity of the assay, an antiserum against whole hCG was purified on a column of hCG beta carboxyl-terminal peptide (residues 123-145) covalently linked to Sepharose 4B. The antibody (N101S) thus prepared showed a weak cross-reactivity with human LH in an assay using hCG-enzyme conjugate, but the slight cross-reactivity was virtually avoided when an hCG beta carboxyl-terminal peptide was used as a peptide in the enzyme conjugate. N101S antibody was compared with antiserum (B1B) directed against a carboxyl-terminal peptide (123-145). In hCG measurement N101S gave about 30 times higher sensitivity than B1B, although the former antibody was less sensitive to carboxyl-terminal peptides of hCG beta. The enzyme immunoassay using a combination of N101S antibody and a carboxyl-terminal peptide (130-145)-enzyme conjugate was able to detect as little as 0.25 mIU of hCG without the interference of LH. The performance and validity of this assay were comparable to those of conventional radioimmunoassay.

Human subtilisin-like proprotein convertase, PACE4 (SPC4) gene expression is highly regulated through E-box elements in HepG2 and GH4C1 cells.

PACE4 (SPC4) is a member of the mammalian subtilisin-like proprotein convertase (SPC) family, which participates in maturation of precursor proteins. PACE4 is expressed at high levels in the anterior pituitary, central nervous system, the developing olfactory bulb, heart, and liver. Recently, we determined the gene structure of human PACE4. [Tsuji et al. (1997) J. Biochem. 122, 438-452]. The 5'-flanking region of PACE4 gene contains 12 E-boxes (E1 to E12) within 1 kb upstream of the transcription initiation site. To examine the function of these E-box elements in the regulation of PACE4 expression, deletion and mutation constructs of the 5'-flanking region were ligated to the luciferase gene and analyzed for promoter activity in HepG2 and GH4C1 cells, which express PACE4 at high level. Some differences were observed in the activity of each promoter construct between HepG2 and GH4C1 cells, although the overall profiles of activity for the promoter fragment series were similar regardless of cell type. We showed that the basal promoter activity of the PACE4 gene is first determined by sequences lying between -315 and -1 bp and further regulated by positive and negative elements in the upstream region. Site-directed mutagenesis of E-boxes in these regulatory elements showed that the E10 E-box act as positive regulator, whereas an E-box cluster (E4-E9) acts as a negative regulator in both cells. E2 E-box acts as a positive regulator only in HepG2 cells. Other E-boxes (E1, E3, and E12) had no effect on the promoter activity. These results indicate that E-box elements play a critical role in controlling PACE4 expression in HepG2 and GH4C1 cells and that PACE4 expression is regulated by a mechanism distinct from that of other SPC family proteases.

Japanese encephalitis vaccine (inactivated, BIKEN) in U.S. soldiers: immunogenicity and safety of vaccine administered in two dosing regimens.

The safety and immunogenicity of Japanese encephalitis (JE) vaccine (Nakayama strain, monovalent / BIKEN) was studied in 538 U.S. soldiers in 1990. Three doses of vaccine from three consecutively manufactured lots were given on days 0, 7, and either 14 or 30. Serum for antibody determination was drawn at months 0, 2, and 6. Japanese encephalitis plaque reduction neutralization tests were performed by three laboratories on each specimen. Five hundred twenty-eight (98%) participants completed the immunization series. All recipients without antibody before immunization developed neutralizing antibody against JE virus. There were no differences in geometric mean titer among the three test lots at months 2 and 6. Soldiers who received the third dose on day 30 had higher titers at both time points. Antibody to yellow fever had no significant effect on immune response to vaccine. Conclusions drawn from analysis of serologic data from the three labs were nearly identical. Symptoms were generally limited to mild local effects and were reduced in frequency with each subsequent does in the series (21% to 11%; P < 0.0001). Generalized symptoms were rare (e.g., fever = 5%) with no reported cases of anaphylaxis.

Thoracoscopic treatment of primary chylopericardium.

The development of video-assisted thoracoscopy has changed the practice of thoracic surgery. This report describes a 20-year-old man who successfully underwent thoracoscopic thoracic duct ligation and partial pericardiectomy for primary chylopericardium. We recommend a right-sided thoracoscopic approach to the thoracic duct in the lower thoracic cavity.

Diverticulum of the superior vena cava.

A 14-year-old girl underwent operation with the diagnosis of diverticulum of the superior vena cava. Microscopic findings revealed a diverticulum with venous architecture. This represents a rare case of a giant diverticulum of the superior vena cava.

Frequency and operative correction of aortic insufficiency associated with ventricular septal defect.

Frequency and surgical results of aortic cusp prolapse and aortic regurgitation (AR) associated with ventricular septal defect (VSD) were studied. One hundred thirty-six consecutive patients with type I and II VSD according to Kirklin and associates' classification were divided into two groups; group A included 50 patients with type Ia VSD without the conal muscular rim, and group B included 86 patients with type Ib VSD with the conal muscular rim, or with type II VSD. Aortic cusp prolapse was detected in 74% of group A patients and 29% of group B patients. The aortic cusp prolapse correlated negatively with preoperative left-to-right shunt ratio, mean pulmonary artery pressure, and pulmonary-to-systemic pressure ratio in both groups. In group B patients, the smaller the VSD, the higher the frequency of aortic cusp prolapse, especially when less than 4 mm. Aortic regurgitation was observed in 44% of group A patients and 24% of group B patients. All 20 patients with first grade AR underwent VSD closure, 11 with second or third grade AR underwent VSD closure plus valvuloplasty, and 1 with third grade AR underwent aortic valve replacement. Postoperative persistent AR occurred in 8 out of 32 patients, and correlated positively (p < 0.01) with the preoperative grade of AR and the number of plication stitches in both groups.