The peptidoglycan of bacterial cell walls is synthesized in a reaction mechanism in which a membrane-bound lipid serves as a carrier for the sugars and amino acids that eventually become part of the peptidoglycan.' One intermediate in the reaction cycle is represented as lipid-P-P-disaccharide-pentapeptide. This intermediate is common to a number of different bacteria. However, its disaccharidepentapeptide moiety (GlcNAc-MurNAc-L-ala-D-glu-L-lys-D-ala-D-ala)I becomes modified in various ways in different organisms.3 The modified disaccharidepeptide moiety is then transferred to an endogenous acceptor with introduction of a new unit into a growing peptidoglycan.The purpose of the present paper is to report that lipid-P-P-disaccharide-pentapeptide has been obtained in a highly purified form from Micrococcus lysodeikticus, and that the nature of the lipid moiety of this intermediate has been elucidated.Soon after the discovery of the occurrence of a lipid intermediate in peptidoglycan synthesis, a lipid carrier in the biosynthesis of the lipopolysaccharide of the cell wall of Salmonella was described.4' I The structure of this substance is reported in a paper by Wright et al-'Isolation of Lipid-P-P-Disaccharide-Pentapeptide from M. lysodeikticus.-Largescale preparation of the lipid intermediate was carried out using techniques essentially the same as those described previously.7 Particulate enzyme was prepared after lysozyme lysis of cells of M. lysodeikticus8 and incubated with the uridine nucleotide substrates to generate the lipid intermediate. The incubation mixture contained 1000 ml of particulate enzyme prepared in 0.05 M Tris-HCl buffer, pH 7.4, and 0.01 M MgCl2, 1.7 ml of 12 mM UDP-MurNAc-pentapeptide (containing 8.7 X 106 cpm of C14-lysine),9 8 ml of 10 mM UDP-GlcNAc and 30 ml of 5 per cent deoxycholate. Enzyme prepared after lysozyme lysis of cells of M. lysodeikticus is able to synthesize the lipid intermediates, but is unable to utilize these intermediates for peptidoglycan synthesis. Deoxycholate was included in the incubation mixture in order to minimize the addition of endogenous glycine to the lipid intermediate.8After incubation at 200 for 30 minutes, the mixture was extracted with 1 liter of nbutanol:6 M pyridinium acetate (2:1), pH 4.2, followed by two extractions with the same volume of n-butanol. The combined extracts were back-washed with 2 liters of water and then taken to dryness and redissolved in 50 ml of chloroform: methanol (1:1). The yield was 3 X 106 cpm (34%, based on UDP-MurNAc-,C4-pentapeptide).The extract was placed on a column of O-(diethylaminoethyl) cellulose (DEAEcellulose) (2.5 X 25 cm) and eluted as previously described.7 The radioactive eluate obtained with methanol:6 M pyridinium acetate, pH 4.2 (1: 1), was pooled 1878

Oral hypofunction is a new concept that addresses the oral function of older adults. Few studies have investigated the relationship between oral hypofunction and general health conditions such as frailty, sarcopenia, and mild cognitive impairment. This paper explores these relationships in a large-scale, cross-sectional cohort study. The relationships of oral hypofunction with frailty, sarcopenia, and mild cognitive impairment were examined using data from 832 individuals who participated in the 2018 health survey of the residents of Tarumizu City, Kagoshima Prefecture, Japan. Individuals with frailty, sarcopenia, and mild cognitive impairment had significantly higher rates of oral hypofunction. Frailty was independently associated with deterioration of the swallowing function (odds ratio 2.56; 95% confidence interval, 1.26–5.20), and mild cognitive impairment was independently associated with reduced occlusal force (odds ratio 1.48; 95% confidence interval, 1.05–2.08) and decreased tongue pressure (odds ratio 1.77; 95% confidence interval, 1.28–2.43). There was no independent association found between sarcopenia and oral function. In conclusion, early intervention for related factors such as deterioration of the swallowing function in frailty, reduced occlusal force, and decreased tongue pressure in mild cognitive impairment could lead to the prevention of general hypofunction in older adults.

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Key Results1. Among a 787-patient cohort with confirmed COVID-19, three chest radiograph scores (BRIXIA, RALE, and percent opacification) all had good interrater reliability with intraclass correlations of 0.87, 0.86, and 0.72 respectively.2. Radiograph scores predicted intensive care unit (ICU) admission or death after COVID-19 diagnosis. A 50%-75% opacification (compared to 0%-25%) associated with a 2.2-fold increase in these outcomes among those eligible for ICU care after adjustment for clinical risk scoring. SummaryBRIXIA, RALE, and percent opacification produced reliable and reproducible COVID-19 chest radiograph severity scores that improved accuracy for predicting adverse outcomes when incorporated into ISARIC-4C mortality and NEWS2 clinical scoring systems. I n p r e s s ResultsAdmission chest radiographs of 50 patients (mean age, 74 years +/-16 [sd], 28 men) were scored by all 3 radiologists, with good inter-rater reliability for all scores (ICCs (95% CIs) of for RALE 0.87 (0.80, 0.92), BRIXIA 0.86 (0.76, 0.92), and percentage opacification 0.72 (0.48, 0.85)). Of 751 patients with chest radiograph, those with >75% opacification had a median time to ICU admission or death of just 1-2 days. Among 628 patients with data (median age 76 years (IQR 61 -84), and 344 were men), 50-75% opacification increased risk of ICU admission or death by twofold (1.6 -2.8), and over 75% opacification by 4 fold (3.4 -4.7), compared to a 0-25% opacification when adjusted for NEWS2 score. ConclusionBRIXIA, RALE, and percent opacification scores all reliably predicted adverse outcomes in SARS-CoV-2.

A genomic library of Clostridium septicum NCTC547 strain was made in Escherichia coli by means of lambda gt10. The DNA insert of a hemolysin-positive (Hly+) lambda-clone was transferred into pUC19. The resulting plasmid, pCS21, confers a Hly+ phenotype on E. coli. Crude lysates of E. coli (pCS21) possessed a strong lytic activity on human erythrocytes and also a lethal effect on mice, characteristic of an alpha toxin. Nucleotide sequence analysis revealed that the insert DNA (5.2 kb) in pCS21 included at least one open reading frame of 1380 bp. The coding frame for hemolysin was predicted to be 1329 bp in size and to encode a protein of 49.8 kDa. It coincided with the molecular mass (48 kDa) of the alpha toxin secreted by C. septicum. Taken together, the data indicated that plasmid pCS21 indeed encoded an alpha toxin gene of C. septicum.