High SARS-CoV-2 Seroprevalence after Second COVID-19 Wave (October 2020-April 2021), Democratic Republic of the Congo.

Serologic surveys are important tools for estimating the true burden of COVID-19 in a given population. After the first wave of SARS-CoV-2 infections, a household-based survey conducted in Kinshasa, Democratic Republic of the Congo, estimated >292 infections going undiagnosed for every laboratory-confirmed case. To ascertain the cumulative population exposure in Kinshasa after the second wave of COVID-19, we conducted a prospective population-based cross-sectional study using a highly sensitive and specific ELISA kit. The survey included 2,560 consenting persons from 585 households; 55% were female and 45% male. The overall population-weighted, test kit-adjusted SARS-CoV-2 seroprevalence was 76.5% (95% CI 74.5%-78.5%). The seroprevalence was 4-fold higher than during the first wave, and positivity was associated with age, household average monthly income, and level of education. Evidence generated from this population-based survey can inform COVID-19 response, especially vaccination campaign strategies in the context of vaccine shortages and hesitancy.

Multi-size spheroid formation using microfluidic funnels.

We present a microfluidic platform for automatic multi-size spheroid formation within constant volume hanging droplets (HDs) from a single inlet loading of a constant cell concentration. The platform introduces three technological improvements over the existing spheroid formation platforms: 1) cell seeding control is achieved by enrichment of a cell solution rather than dilution; 2) cell seeding in each HD is fully independent and pre-programmable at the design stage; 3) the fabricated chip operates well using a hydrophobic PDMS surface, ensuring long-term storage possibility for device usage. Pre-programmed cell seeding densities at each HD are achieved using a "microfluidic funnel" layer, which has an array of cone-shaped wells with increasing apex angles acting as a metering unit. The integrated platform is designed to form, treat, stain, and image multi-size spheroids on-chip. Spheroids can be analyzed on-chip or easily transferred to conventional well plates for further processing. Empirically, enrichment factors up to 37× have been demonstrated, resulting in viable spheroids of diameters ranging from 230-420 µm and 280-530 µm for OV90 and TOV112D cell lines, respectively. We envision that microfluidic funnels and single inlet multi-size spheroid (SIMSS) chips will find broad application in 3D biological assays where size-dependent responses are expected, including chemoresponse assays, photodynamic therapy assays, and other assays involving drug transport characterization in drug discovery.

Micro-dissected tumor tissues on chip: an ex vivo method for drug testing and personalized therapy.

In cancer research and personalized medicine, new tissue culture models are needed to better predict the response of patients to therapies. With a concern for the small volume of tissue typically obtained through a biopsy, we describe a method to reproducibly section live tumor tissue to submillimeter sizes. These micro-dissected tissues (MDTs) share with spheroids the advantages of being easily manipulated on-chip and kept alive for periods extending over one week, while being biologically relevant for numerous assays. At dimensions below ~420 µm in diameter, as suggested by a simple metabolite transport model and confirmed experimentally, continuous perfusion is not required to keep samples alive, considerably simplifying the technical challenges. For the long-term culture of MDTs, we describe a simple microfluidic platform that can reliably trap samples in a low shear stress environment. We report the analysis of MDT viability for eight different types of tissues (four mouse xenografts derived from human cancer cell lines, three from ovarian and prostate cancer patients, and one from a patient with benign prostatic hyperplasia) analyzed by both confocal microscopy and flow cytometry over an 8-day incubation period. Finally, we provide a proof of principle for chemosensitivity testing of human tissue from a cancer patient performed using the described MDT chip method. This technology has the potential to improve treatment success rates by identifying potential responders earlier during the course of treatment and providing opportunities for direct drug testing on patient tissues in early drug development stages.

Minor H antigen matches and mismatches are equally distributed among recipients with or without complications after HLA identical sibling renal transplantation.

Studies of the effect of minor H antigen mismatching on the outcome of renal transplantation are scarce and concern mainly single center studies. The International Histocompatibility and Immunogenetics Workshops (IHIW) provide a collaborative platform to execute crucial large studies. In collaboration with 16 laboratories of the IHIW, the role of 15 autosomal, 10 Y-chromosome encoded minor H antigens and 3 CD31 polymorphisms, was investigated in relation to the incidence of renal graft rejection and graft loss in 444 human leukocyte antigens (HLA)-identical sibling renal transplantations. Recipient and donor DNA samples were genotyped for the minor H antigens HA-1, HA-2, HA-3, HA-8, HB-1, ACC-1, ACC-2, SP110, PANE1, UGT2B17, C19Orf48, LB-ECGF-1, CTSH, LRH-1, LB-ADIR and HY. The correlation between minor H antigen mismatch and the primary outcome graft rejection or graft loss was statistically analyzed. The incidence of rejection was very low and no correlation was observed between one or more minor H antigen mismatch(es) and a rejection episode (n = 36), of which only eight resulted in graft loss. In summary, in our study cohort of 444 renal transplants, mismatching for neither autosomal nor HY minor H antigens correlate with rejection episodes or with graft loss.

[Susceptibility genes, HLA and diabetic retinopathy in the Algerian population]. / Gènes de susceptibilité HLA et rétinopathie diabétique chez la population algérienne.

BACKGROUND: Diabetic retinopathy (DR) is the most frequent microvascular complication of type I diabetes (T1D). Some well-controlled type I diabetics may develop DR, while other poorly-controlled diabetics do not develop DR. This might be explained by certain susceptibility genes or protective genes. The purpose of our study is to search for any association between the HLA class I and II markers and DR in the Algerian population. PATIENTS AND METHODS: This study was carried out in 52 T1D subjects with and without DR compared to 140 healthy controls. HLA typing was performed using the "microlymphocytotoxicity" technique. RESULTS: The frequency of HLA-A29 and HLA-DR9 antigens is higher in T1D with DR compared to T1D without DR and to controls with frequencies of HLA-A29 (59.26% vs. 0%, OR=∞, pc=4.6×10(-7)), (59.26% vs. 5.66%, OR=24.24, pc=7.6×10-10) and HLA-DR9 (29.63% vs. 0%, OR=∞, pc=1.310(-3)), (29.63% vs. 4.29%, OR=9.40, pc=7.010(-5)) respectively. However, the frequency of HLA-B49 antigen is significantly lower in T1D with DR than in T1D without DR (3.7% vs. 28%, OR=0.10, pc=8.8×10(-3)) and compared to controls (3.7% vs. 22.64%, OR=0.13, pc=0.011). CONCLUSION: HLA-A29 and HLA-DR9 antigens are probably markers of susceptibility for DR while HLA-B49 antigen is probably associated with a protective effect in the Algerian population.

A novel HLA-A*11 allele, A*1118, identified by sequence-based typing.

The novel allele A*1118 shows three nucleotide differences with A*110101, resulting in three amino acid changes at positions 70, 74 and 90 of the mature protein.

Identification of the new allele B*3556 and extension of the sequences of B*4420 and B*4427.

A new B*35 allele, B*3556, was identified in a Caucasian individual. Direct sequencing of exons 2 and 3 revealed that B*3556 was identical to B*3503, except for a nucleotide substitution from G to A at position 302, which causes a change from AGC to AAC at codon 101. This results in an amino acid change from Ser to Asn at position 77 of the mature protein. Two B*44 alleles, B*4420 and B*4427, were detected in a liver recipient and in an unrelated bone marrow donor, respectively, and their sequences were confirmed. The sequences of exons 1,4 and 5 were also elucidated; for B*4420, these exons proved to be identical to those of B*440201. The sequence of exon 5 of B*4427 showed three mismatches with that of B*440201, implicating that it probably arose from B*440201 by allele conversion with an allele of the B*15, 45, 46, 49 or 50 allele group. Elucidation of the sequence of introns 3 and 4 indicated that the breakpoint for allele conversion has to be located in intron 3 or exon 4.

Results of inspection and cleaning of two radionuclide air-sampling systems based on the requirements of ANSI/HPS N13.1-1999.

The Pacific Northwest National Laboratory inspected and cleaned two radionuclide air-sampling systems that continuously monitor radioactive air emissions from research and development facilities. The inspection and cleaning was performed to evaluate effective methods and potential cost impacts of maintenance requirements in the revised American National Standard Institute standard Sampling and Monitoring Releases of Airborne Radioactive Substances from the Stacks and Ducts of Nuclear Facilities. The standard requires at least annual inspections of sampling systems followed by cleaning if deposits are visible. During 2001 and 2002, inspections were performed leaving the sampling systems in place and inserting videoscope cables into different access points to allow viewing of the inside and outside of sampling manifolds and transport lines. Cleaning was performed on one of the systems by disconnecting and extracting the sampling manifold, then washing it with de-ionized water and scrub brushes. The wash water was analyzed for radioactivity and solids. Results of the inspection showed greater deposition in one of the systems than would be expected by a High Efficiency Particulate Air (HEPA) filtered exhaust stream, possibly due to accumulation of dust from a short period when unfiltered air was exhausted from construction areas. The second system was also downstream of HEPA filters and appeared much cleaner. The videoscope was a useful and cost-effective tool and provided a better view than could be obtained with the naked eye. However, because even small amounts of deposition were made visible with the videoscope, clarification is needed in defining when probe washing is merited, particularly in existing sampling systems whose design is not conducive to easy removal and cleaning.

Sequence-based typing identifies a novel HLA-DPB1 allele, DPB1*9601.

In this report we describe the identification of a novel HLA-DPB1 allele, DPB1*9601, found in a Caucasian individual sample named ucla#356. The new allele was detected in the DNA of ucla#356 during routine HLA sequence-based typing (SBT) of samples participating in the UCLA International HLA DNA Exchange (number 55) for HLA DNA Proficiency Testing. DPB1*9601 was identical to DPB1*3901 except for a single nucleotide substitution 'G'-->'C' in previously constant position 277 (position 177, respectively, counting only exon 2). This nucleotide change causes an amino acid substitution from aspartic acid in DPB1*3901 to histidine at codon 64 in the novel allele. This new allele has been submitted to the EMBL database and has been assigned the accession number AJ514871. The WHO Nomenclature Committee has officially assigned the name DPB1*9601.

Identification of HLA-A*0249 and confirmation of A*2615.

A new HLA-A*02 allele, A*0249, and the confirmatory sequence of A*2615 are here reported. Both alleles were detected by irregular patterns during routine molecular typing. A*0249 showed two mismatches with A*02011 in exon 3 at positions 538 and 539, changing amino acid 156 from leucine to arginine. Remarkable was a mismatch in exon 4, at position 779, where the C of A*0201 was changed to A in A*0249. As A*0209 has the same substitution, A*0249 may have arisen from A*0209. The A*2615 allele was identical to A*2601 in exons 2 and 3 except for a single nucleotide difference at nucleotide position 180, changing codon 36 from phenylalanine to leucine. This is a unique amino acid change, as in all the class I alleles a phenylalanine was identified at this position.

Effect of aeration during cell growth on ketone reactions by immobilized yeast.

The effect of aeration during cell growth on the subsequent reduction of 2-hexanone and 2-octanone by yeast cells entrapped in calcium alginate beads was studied. The reactions were conducted using 2-propanol as a sacrificial substrate to regenerate the cofactor NAD(H), and a mixture of (S)- and (R)-alcohols was produced. The use of strictly aerobic conditions when growing the cells resulted in the highest initial reaction rates, as well as the production of only a single product (i.e., the enantiomeric excess of the (S)-alcohols was 1.0). However, initial reaction rates decreased proportionally with fermentation time regardless of whether the yeast were grown aerobically or under both aerobic and anaerobic conditions. The data also suggest that it is the aerobic (or anaerobic) condition, rather than the cell growth phase, which is responsible for the results seen.

Kinetics of protein depletion in rat bronchoalveolar lavage fluid following in vitro exposure to nitrogen dioxide.

Upon inhalation, nitrogen dioxide (NO(2)), a strong oxidizing agent, first comes into contact and reacts with the fluids lining the airways of the respiratory tract. These respiratory tract lining fluids (RTLF) form a barrier between the inhaled toxic pollutant and the epithelium which protects the underlying tissue from inflammation. Proteins, mainly albumin, and antioxidants are the major components of the RTLF. Many studies have utilized human blood plasma to study the interaction of an extracellular fluid with ozone. In this study, we used bronchoalveolar lavage fluids (BALF) as a more specific surrogate for rat RTLF, and we utilized the native fluorescence as a marker to investigate the depletion kinetics of naturally-occurring protein following exposure to NO(2) in a controlled flow reactor system. We also studied the depletion kinetics of albumin in a buffered salt solution. The results indicated that: (1) the decay in fluorescence was linearly dependent on the concentration of NO(2), indicating that protein oxidation was first order with respect to NO(2) concentration in both BALF and in buffered albumin solution; (2) the depletion kinetics of protein in BALF was non-linear with respect to substrate concentration; (3) the rate of protein depletion was much slower in BALF than in a buffered solution of albumin, suggesting that the presence of antioxidants in BALF protected proteins from being oxidized by NO(2); and (4) whereas the addition of ascorbic acid to buffered albumin solution significantly attenuated albumin depletion, the addition of glutathione had no effect. This suggested that the reaction rate constant of ascorbic acid was considerably higher than that of glutathione.

[Portal vein thrombosis and hereditary protein C deficiency. Presentation of a case and review of the literature]. / Thrombose portale et déficit héréditaire en protéine C. Présentation d'un cas et revue de la littérature.

The authors report the case of a 51 year-old man, without any personal or familial history of thromboembolism, presenting with abdominal pain. Portal vein thrombosis was demonstrated by ultrasonography and arteriography. The patient had neither esophageal varices or congestive gastropathy. No cause for portal vein thrombosis was detected. Type I protein C deficiency was demonstrated in this patient as well as in his asymptomatic sister. The presence of a (fortuitously?) associated increase in platelet aggregability initially led to a trial regimen of aspirin (300 mg per day); abdominal pain resolved, and a partial regression of portal vein thrombosis was demonstrated on ultrasonograms six months later; no further complications occurred during the 4-year follow-up period. The 13 previously published cases of protein C deficiency-associated portal vein thrombosis are reviewed.

Thoracic and lower cervical spine involvement in a case of rheumatoid arthritis.

The authors report a case of spine involvement in a severe case of rheumatoid arthritis treated with corticosteroids. First, the patient developed acute back pain, related to costovertebral joints arthritis at levels T9-T10. Then, neck pain and cord involvement yielded to diagnosis of cervical interapophyseal joints arthritis; there was a C5-C6 subluxation which necessitated surgical treatment. The conjunction of these two rheumatoid localizations is an uncommon feature. Study by the CT scan is valuable when rheumatoid arthritis of the spine is suspected. Lower cervical spine subluxation, even severe, may be well tolerated. Surgery is necessary when there is medullary involvement.

Costo-vertebral arthropathy. Diagnostic and therapeutic value of arthrography.

The authors report five cases of costo-vertebral (CV) joint arthropathies, without osteo-arthritis. The patients had referred pain, with misleading irradiations to the loin or to the abdomen, so that the diagnosis was difficult. The authors have performed in these five cases an arthrography of T11 and T12 CV joints. In every case, the arthrography triggered pain at one of these levels. The arthrography was followed by intra-articular injection of corticosteroid derivative in the symptomatic joint. In these cases, this procedure was a valuable diagnostic and therapeutic method. CV joint arthropathies are frequent, but often lead to incorrect diagnosis. These preliminary results encourage further use of this procedure in CV arthropathies.