Outcomes of Kidney Transplant During the SARS-CoV-2 Pandemic at 1 Center in Poland.

The SARS-CoV-2 pandemic has significantly affected the number of transplanted organs worldwide. The rules and restrictions related to transplantation activities in Poland are included in the updated guidelines of the Polish Organizational and Coordination Centre for Transplantation. Our clinic faces the same problems as the rest of the hospitals in the country. Not only are the number of recipients falling, but there are also numerous restrictions concerning, among other things, qualification of donors and recipients and even preparation of centers for long-term care in the event of infection of organ recipients with the SARS-CoV-2 virus. Statistics showed, after an initial fall in the number of kidneys transplanted, a temporary normalization during the summer months, only to record a fall again with an increase in new cases of COVID-19. A total of 29 kidneys were transplanted at our center between March and December 2020. Kidney transplantation is not only linked to the operation itself, but also to the follow-up care of the recipients. Reduced immunity among recipients due to immunosuppressive treatment as well as comorbidities among recipients contribute to this group being at increased risk of symptomatic SARS-CoV-2 infection. The number of cases of SARS-CoV-2 infection among kidney transplant recipients at our center was 7, of which we recorded 2 deaths due to COVID-19 in the period after kidney transplant. Postoperative complications probably related to previous SARS-CoV-2 infection occurred in 1 patient.

Stereospecificity and other properties of a novel secondary-alcohol-specific alcohol dehydrogenase.

NAD-dependent alcohol dehydrogenase from the methanol-grown Methylcoccus sp. CRL M1 (type I membrane), Methylosinus trichosporium OB3b (type II membrane), Methylobacterium organophillum CRL 26 (type II membrane, facultative methylotroph). Pseudomonas sp. ATCC 21439, and Pichia pastoris Y-55 are secondary-alcohol-specific and that from P. pastoris Y-7556 is not. This novel secondary-alcohol-specific alcohol dehydrogenase (secondary-alcohol dehydrogenase) has been purified from methanol-grown Pseudomonas sp. ATCC 21439. Secondary-alcohol dehydrogenase shows a single protein band on acrylamide gel electrophoresis and has a molecular weight of 95000. It consists of two subunits of Mr 48000 daltons and two atoms of zinc per molecule of enzyme protein. It oxidizes secondary alcohols, notably 2-propanol and 2-butanol. Primary alcohols are not oxidized. The pH and temperature optima for secondary-alcohol dehydrogenase are 8--9, and 30--35 degrees C, respectively. The activation energy calculated is 82.8 kJ. Secondary-alcohol dehydrogenase also catalyzes the reduction of methyl ketones to their corresponding 2-alcohols in the presence of NADH (a reverse reaction). The Km values at 25 degrees C in the forward reaction for 2-butanol, (2R)-(-)-butan-2-ol, and NAD, and in the reverse reaction for 2-butanone and NADH are 2.5 x 10(-4) M, 1.6 x 10(-4) M, 11 x 10(-5) M, 1.98 x 10(-4) M, and 2.1 x 10(-6) M, respectively. The secondary-alcohol dehydrogenase activity was inhibited by metal-chelating agents and by strong thio reagents such as p-hydroxymercuribenzoate and 5,5'-dithiobis(2-nitrobenzoic acid). The substrate specificity, and mobility on gel electrophoresis of secondary-alcohol dehydrogenase and primary-alcohol dehydrogenases are compared. Secondary-alcohol dehydrogenase oxidizes preferentially the (-)-2-butanol. This is different from primary-alcohol dehydrogenase from bakers' yeast which oxidizes only the (+)-2-butanol. This may be explained in terms of the structure of the enzymes.

Substrate specificity and stereospecificity of nicotinamide adenine dinucleotide-linked alcohol dehydrogenases from methanol-grown yeasts.

Nicotine adenine dinucleotide-linked primary alcohol dehydrogenase and a newly discovered secondary alcohol dehydrogenase coexist in most strains of methanol-grown yeasts. Alcohol dehydrogenases from methanol-grown yeasts oxidize (--)-2-butanol preferentially over its (+) enantiomorph. This is substantially different from alcohol dehydrogenases from bakers' yeast and horse liver.

Microbial oxidation of gaseous hydrocarbons: production of alcohols and methyl ketones from their corresponding n-alkanes by methylotrophic bacteria.

Cell suspensions of methane-utilizing bacteria oxidized n-alkanes (propane, butane, pentane, and hexane) to their corresponding alcohols and methyl ketones. The product alcohols and methyl ketones accumulated extracellularly. Methanol-grown cells of methane-utilizing bacteria did not oxidize n-alkanes. The product primary alcohol was detected in a cell-free system but only in a trace amount in the whole cell system due to further oxidation. The optimum conditions for in vivo formation of secondary alcohol and methyl ketone from n-alkanes were compared between two distinct types of C1-utilizing microbes: Methylococcus capsulatus M1 (type I membrane) and Methylosinus trichosporium OB3b (type II membrane). The production of acetone or 2-butanone from n-alkanes ceased after 3 h of incubation for strain OB3b and 5 h for strain M1. The amount of these methyl ketones did not decline during 30 h of incubation. The optimum pH for the in vivo production of methyl ketones from n-alkanes by both strains was around 7.0. However, secondary alcohols were accumulated at higher amounts around pH 6.0. The optimum temperature for the in vivo production of methyl ketones from n-alkanes was around 40 degrees C for strain M1 and around 30-35 degrees C for strain OB3b. Higher accumulation of secondary alcohol was detected at 30-40 degrees C for strain M1 and 25 degrees C for strain OB3b. The alkane hydroxylation enzyme was located in the cell-free particulate fraction precipitated between 10 000 and 40 000 X g centrifugation. The yield of primary and secondary alcohols from n-alkane in the cell-free system was about equal. Evidence obtained indicates that the hydroxylation of n-alkanes (both terminal and subterminal oxidations) is also catalyzed by the methane hydroxylation - alkene epoxidation enzyme system.

Microbial oxidation of gaseous hydrocarbons: production of methyl ketones from their corresponding secondary alcohols by methane- and methanol-grown microbes.

Cultures of methane- or methanol-utilizing microbes, including obligate (both types I and II) and facultative methylotrophic bacteria, obligate methanol utilizers, and methanol-grown yeasts were isolated from lake water of Warinanco Park, Linden, N.J., and lake and soil samples of Bayway Refinery, Linden, N.J. Resting-cell suspensions of these, and of other known C1-utilizing microbes, oxidized secondary alcohols to their corresponding methyl ketones. The product methyl ketones accumulated extracellularly. Succinate-grown cells of facultative methylotrophs did not oxidize secondary alcohols. Among the secondary alcohols, 2-butanol was oxidized at the highest rate. The optimal conditions for in vivo methyl ketone formation were compared among five different types of C1-utilizing microbes. Some enzymatic degradation of 2-butanone was observed. The product, 2-butanone, did not inhibit the oxidation of 2-butanol. The rate of the 2-butanone production was linear for the first 4 h of incubation for all five cultures tested. A yeast culture had the highest production rate. The optimum temperature for the production of 2-butanone was 35 degrees C for all the bacteria tested. The yeast culture had a higher temperature optimum (40 degrees C), and there was a reasonably high 2-butanone production rate even at 45 degrees C. Metal-chelating agents inhibit the production of 2-butanone, suggesting the involvement of metal(s) in the oxidation of secondary alcohols. Secondary alcohol dehydrogenase activity was found in the cell-free soluble extract of sonically disrupted cells. The cell-free system requires a cofactor, specifically nicotinamide adenine dinucleotide, for its activity. This is the first report of a nicotinamide adenine dinucleotide-dependent, secondary alcohol-specific enzyme.