An evaluation of a pilot of daily testing of SARS-CoV-2 contacts in acute hospital and ambulance trusts in England.

OBJECTIVES: Healthcare worker (HCW) SARS-CoV-2 contacts in England have been required to quarantine, creating staff shortages. We piloted daily contact testing (DCT) to assess its feasibility as an alternative. STUDY DESIGN: Observational service evaluation. METHODS: We conducted an observational service evaluation of 7-day DCT using antigen lateral flow devices (LFDs) at four acute hospital trusts and one ambulance trust in England. Mixed methods were used, using aggregate and individual-level test monitoring data, semi-structured interviews, and a survey of eligible contacts. RESULTS: In total, 138 HCWs were identified as contacts of a confirmed SARS-CoV-2 case. Of these, 111 (80%) consented to daily LFD testing, of whom 82 (74%) completed the required programme without interruption and 12 (11%) completed with interruption. Fifty-eight participants (52%) and two non-participants (7.4%) completed the survey. In total, 28 interviews were conducted with participants, site and infection control leads, and union representatives. One participant tested positive on LFD and polymerase chain reaction (PCR) test. Three participants tested positive on PCR but not LFD. DCT was well-accepted by trusts and staff. Participants reported no relaxation of their infection prevention and control behaviours. No incidents of transmission were detected. An estimated 729 potential days of work absence were averted. CONCLUSIONS: DCT can be acceptably operated in a healthcare setting, averting quarantine-related work absences in HCW SARS-CoV-2 contacts.

Beyond knockouts: the International Knockout Mouse Consortium delivers modular and evolving tools for investigating mammalian genes.

The International Knockout Mouse Consortium (IKMC; http://www.mousephenotype.org ) has generated mutations in almost every protein-coding mouse gene and is completing the companion Cre driver resource to expand tissue-specific conditional mutagenesis. Accordingly, the IKMC has carried out high-throughput gene trapping and targeting producing conditional mutations in murine embryonic stem cells in more than 18,500 genes, from which at least 4900 mutant mouse lines have been established to date. This resource is currently being upgraded with more powerful tools, such as visualization and manipulation cassettes that can be easily introduced into IKMC alleles for multifaceted functional studies. In addition, we discuss how existing IKMC products can be used in combination with CRISPR technology to accelerate genome engineering projects. All information and materials from this extraordinary biological resource together with coordinated phenotyping efforts can be retrieved at www.mousephenotype.org . The comprehensive IKMC knockout resource in combination with an extensive set of modular gene cassettes will continue to enhance functional gene annotation in the future and solidify its impact on biomedical research.

Hormonal stimulation in the exocrine pancreas results in coordinate and anticoordinate regulation of protein synthesis.

24-h intravenous caerulein infusion studies in the rat were combined with in vitro amino acid incorporation studies followed by high-resolution separation of proteins by two-dimensional isoelectric focusing and SDS gel electrophoresis to study the extent to which persistent changes in the biosynthesis of exocrine pancreatic proteins are regulated by cholecystokinin-like peptides. Beginning in the third hour of optimal hormone infusion at 0.25 microgram kg-1 h-1, changes were observed in the synthetic rates of 12 proteins, which progressed over the course of the 24-h study. Based on coordinate response patterns, exocrine proteins could be classified into four distinct groups. Group I (trypsinogen forms 1 and 2) showed progressive increases in synthetic rates reaching a combined 4.3-fold increase over control levels. Group II (amylase forms 1 and 2) showed progressive decreases in synthesis to levels 7.1- and 14.3-fold lower than control levels, respectively. Group III proteins (ribonuclease, chymotrypsinogen forms 1 and 2, procarboxypeptidase forms A and B, and proelastase 1) showed moderate increases in synthesis, 1.4-2.8-fold, and group IV proteins (trypsinogen 3, lipase, proelastase 2, and unidentified proteins 1-4) did not show changes in synthesis with hormone stimulation. Regulation of protein synthesis in response to caerulein infusion was specific for individual isoenzymic forms in the case of both trypsinogen and proelastase. The ratio of biosynthetic rates of trypsinogen forms 1 + 2 to amylase forms 1 + 2 increased from a control value of 0.56 to 24.4 after 24 h of hormonal stimulation (43.5-fold increase). Biosynthetic rates for an unidentified protein (P23) with an Mr = 23,000 and isoelectric point of 6.2 increased 14.2-fold, and the ratio of synthesis of P23 to amylase 2 increased 200-fold during caerulein infusion. During hormone stimulation the anticoordinate response in the synthesis of pancreatic glycosidases (decreased synthesis) and serine protease zymogens (increased synthesis) explain previous observations that showed little change in rates of total protein synthesis under similar conditions.

Contamination of organic nutrient sources with potentially toxic elements, antibiotics and pathogen microorganisms in relation to P fertilizer potential and treatment options for the production of sustainable fertilizers: A review.

Organic nutrient sources such as farmyard manure, sewage sludge, their biogas digestates or other animal by-products can be valuable fertilizers delivering organic matter to the soil. Currently, especially phosphorus (P) is in the focus of research since it is an essential plant nutrient with finite resources, estimated to last only for some more decades. Efficient utilization of organic P sources in agriculture will help to preserve P resources and thereby has the potential to close nutrient cycles and prevent unwanted P-losses to the environment, one of the major causes for eutrophication of water bodies. Unfortunately, organic P sources usually contain also various detrimental substances, such as potentially toxic elements or organic contaminants like pharmaceuticals as well as pathogenic microorganisms. Additionally, the utilization of some of these substrates such as sewage sludge or animal by-products is legally limited in agriculture because of the potential risk to contaminate sites with potentially toxic elements and organic contaminants. Thus, to close nutrient cycles it is important to develop solutions for the responsible use of organic nutrient sources. The aim of this review is to give an overview of the contamination of the most important organic nutrient sources with potentially toxic elements, antibiotics (as one important organic contaminant) and pathogenic microorganisms. Changes in manure and sewage sludge management as well as the increasing trend to use such substrates in biogas plants will be discussed with respect to potential risks posed to soils and water bodies. Some examples for abatement options by which contamination can be reduced to produce P fertilizers with high amounts of plant available P forms are presented.

[Patterns of prescription and usage of triptans in Alsace (France): misuse is frequent and avoidable]. / Modes de prescription et de consommation des triptans en Alsace: un mésusage fréquent mais évitable.

AIMS: To analyze use of triptans in the Alsace region of France: patients, disorders motivating, doses, analgesics and migraine prophylactics associated treatments, contra-indications. To study major consumers (more than 144 intakes per year) and to determine among them the proportion who suffering from chronic headache. METHOD: Data concerning all prescriptions of triptans and analgesics as well as migraine prophylaxis prescriptions were obtained from the computer databases of five of the French National Health's local health agencies in Alsace, recorded between April 1, 2003 and March 31, 2004. Data about motivating disorders and the clinical context were obtained using a questionnaire sent to prescribers. Data about patients with more than 144 intakes per year were provided by medical advisors of French Health insurance. RESULTS: We founded 20686 users: 92.1 percent used between 0 and 6 intakes per month. 11.5 percent of disorders motivating the prescription that were mentioned by prescribers were for off-label use: tension-type headache 2.7 percent, mixed headache, 8.8 percent. Prescribers declared at least one contra-indication for triptan use for 7.8 percent of patients. Over all, prescriptions were off-label for 16.1 percent of patients. Patients who used more than 144 intakes per year accounted for 1.9 percent of the total number and self-medication accounted for 19.2 percent of all triptan intakes. Half of the patients were suffering from daily chronic headache (chronic migraine in 66 percent). 15.6 percent of these patients presented at least one contraindication (high blood pressure or ischemic disease). All in all we estimate that use of triptan is a misuse for 25 percent to 30 percent of the intakes. Quantities of other analgesics used increased simultaneously with triptan use: on average 65, 119 and 244 Defined Daily Doses (DDD)/person/year for patients who used between 1 and 72, 73 and 144 and more than 144 intakes respectively. On average 35.4 percent (in DDD) of analgesics used were opiates (dextropropoxyphene, codeine, tramadol). This proportion increased simultaneously with triptan use: 58.9 percent for major users. Prophylactic treatment for migraine was used by 27.9 percent of the patients: lack of prophylaxis was a prescriber's choice in 90 percent of the cases. CONCLUSIONS: The high rate of triptan misuse emphasizes the importance of improving prescription of these drugs.

Interaction of gender and body mass index (BMI) reveals evidence of linkage for hypertension in the Framingham Heart Study.

BACKGROUND: Genetic heterogeneity and complex biologic mechanisms of blood pressure regulation pose significant challenges to the identification of susceptibility loci influencing hypertension. Previous linkage studies have reported regions of interest, but lack consistency across studies. Incorporation of covariates, in particular the interaction between two independent risk factors (gender and BMI) greatly improved our ability to detect linkage. RESULTS: We report a highly significant signal for linkage to chromosome 2p, a region that has been implicated in previous linkage studies, along with several suggestive linkage regions. CONCLUSION: We demonstrate the importance of including covariates in the linkage analysis when the phenotype is complex.

Characterization of pepR1, a gene coding for a potential transcriptional regulator of Lactobacillus delbrueckii subsp. lactis DSM7290.

The gene designated pepR1, encoding a potential transcription regulator of Lactobacillus delbrueckii subsp. lactis DSM7290, was identified by sequence similarity of an open reading frame located upstream of the prolidase pepQ orientated in opposite direction. pepQ and pepR1 coding regions are spaced by 152 nucleotides. Upstream of the -35 region of pepQ, a 14-bp palindromic sequence, homologous to the catabolite responsive element, could be identified. The pepRl gene has the potential to encode a protein of 333 amino acids with a calculated molecular mass of 36955 Da and a calculated pl of 5.5. The deduced protein sequence shows significant identity to the catabolite control protein of Bacillus. Co-expression in Escherichia coli was studied with the pepR1-pepQ intergenic region fused to the promoterless beta-galactosidase reporter gene. The pepQ-beta-galactosidase hybrid displayed an enhanced expression in the presence of cloned pepR1.

Umbilical artery catheter position and intraventricular hemorrhage.

A historical cohort study was designed to examine high (T6-T11) versus low (L3-L5) umbilical artery catheter (UAC) positioning as a risk factor for subependymal/intraventricular hemorrhage (SEH/IVH) in very low birthweight infants. High and low UAC groups were similar for mode of delivery, severity of pulmonary disease, weight, gestation, Apgar scores, and air leak. Seventeen of 36 infants with high UACs and 6 of 44 infants with low UACs developed SEH/IVH. The incidence (P less than .001) and severity (P less than .01) of SEH/IVH was significantly greater in the high UAC positioning group. Retrograde arterial flow with or without embolization to the cerebral circulation from high UAC positioning is suggested as a possible cause for the association between high UAC positioning and SEH/IVH.

Use of a truss to maintain inguinal hernia reduction in a very low birth weight infant.

Trusses are not usually used in management of inguinal hernia of the very low birth weight infant. A potential benefit of this therapy is maintenance of hernia reduction, thus delaying operative repair until the infant is larger and healthier. We designed a safe and effective truss with supplies found in most neonatal intensive care units.

Gene expression profiling of human ovarian tumours.

There is currently a lack of reliable diagnostic and prognostic markers for ovarian cancer. We established gene expression profiles for 120 human ovarian tumours to identify determinants of histologic subtype, grade and degree of malignancy. Unsupervised cluster analysis of the most variable set of expression data resulted in three major tumour groups. One consisted predominantly of benign tumours, one contained mostly malignant tumours, and one was comprised of a mixture of borderline and malignant tumours. Using two supervised approaches, we identified a set of genes that distinguished the benign, borderline and malignant phenotypes. These algorithms were unable to establish profiles for histologic subtype or grade. To validate these findings, the expression of 21 candidate genes selected from these analyses was measured by quantitative RT-PCR using an independent set of tumour samples. Hierarchical clustering of these data resulted in two major groups, one benign and one malignant, with the borderline tumours interspersed between the two groups. These results indicate that borderline ovarian tumours may be classified as either benign or malignant, and that this classifier could be useful for predicting the clinical course of borderline tumours. Immunohistochemical analysis also demonstrated increased expression of CD24 antigen in malignant versus benign tumour tissue. The data that we have generated will contribute to a growing body of expression data that more accurately define the biologic and clinical characteristics of ovarian cancers.

Identification of genes associated with platinum drug sensitivity and resistance in human ovarian cancer cells.

Platinum-based chemotherapeutic regimens are ultimately unsuccessful due to intrinsic or acquired drug resistance. Understanding the molecular basis for platinum drug sensitivity/resistance is necessary for the development of new drugs and therapeutic regimens. In an effort to identify such determinants, we evaluated the expression of approximately 4000 genes using cDNA microarray screening in a panel of 14 unrelated human ovarian cancer cell lines derived from patients who were either untreated or treated with platinum-based chemotherapy. These data were analysed relative to the sensitivities of the cells to four platinum drugs (cis-diamminedichloroplatinum (cisplatin), carboplatin, DACH-(oxalato)platinum (II) (oxaliplatin) and cis-diamminedichloro (2-methylpyridine) platinum (II) (AMD473)) as well as the proliferation rate of the cells. Correlation analysis of the microarray data with respect to drug sensitivity and resistance revealed a significant association of Stat1 expression with decreased sensitivity to cisplatin (r=0.65) and AMD473 (r=0.76). These results were confirmed by quantitative RT-PCR and Western blot analyses. To study the functional significance of these findings, the full-length Stat1 cDNA was transfected into drug-sensitive A2780 human ovarian cancer cells. The resulting clones that exhibited increased Stat1 expression were three- to five-fold resistant to cisplatin and AMD473 as compared to the parental cells. The effect of inhibiting Jak/Stat signalling on platinum drug sensitivity was investigated using the Janus kinase inhibitor, AG490. Pretreatment of platinum-resistant cells with AG490 resulted in significant increased sensitivity to AMD473, but not to cisplatin or oxaliplatin. Overall, the results indicate that cDNA microarray analysis may be used successfully to identify determinants of drug sensitivity/resistance and future functional studies of other candidate genes from this database may lead to an increased understanding of the drug resistance phenotype.

Integration of adenovirus type 2 DNA in productively infected cells: results of sequential hybridization experiments.

In human KB cells permissively infected with adenovirus type 2, the high mol. wt. forms of virus DNA have been characterized. These size classes of virus DNA sediment at greater than 100S and 40 to 90S in alkaline sucrose density gradients. Considerable evidence from a series of earlier communications supports the notion that the high mol. wt. forms of virus DNA represent virus DNA sequences covalently linked to cellular DNA. 3H-labelled high mol. wt. adenovirus type 2 DNA from productively infected cells can be shown to hybridize to virus DNA fixed to filters. In the present paper we demonstrate that on alkali elution of the DNA from the filters used in the first step of the hybridization experiment, the labelled DNA re-hybridizes to cellular DNA in the second step of a sequential hybridization experiment. The order of performing the two successive hybridization experiments can be reversed and very similar results are obtained. These data provide conclusive evidence for the covalent linkage of virus and cellular DNA sequences in KB cells productively infected with adenovirus type 2.

High molecular weight virus DNA in KB cells infected with ts mutants of adenovirus type 2 under permissive and non-permissive conditions.

Temperature-sensitive (ts) mutants of adenovirus type 2 (Ad2), which are deficient in virus DNA synthesis at the non-permissive temperature, have been used to investigate whether virus DNA replication is required for the occurrence of high mol. wt. Ad2 DNA (greater than 100S, 50 to 90S) in human cells productively infected with Ad2. The high mol. wt. virus DNA has been previously shown to consist of virus and cellular DNA molecules covalently linked. The present data indicate that after infection with DNA-ts mutants, the production of high mol.wt. virus DNA is much less sensitive to restrictive conditions than the synthesis of unit length (34S) Ad2 DNA. This finding lends further support to the idea that the occurrence of high mol. wt. virus DNA is independent of the synthesis of unit length virus DNA.

Determination of cytarabine and uracil arabinoside in human plasma and cerebrospinal fluid by high-performance liquid chromatography.

A high-performance liquid chromatographic method for the determination of the antineoplastic agent cytarabine and its main metabolite uracil arabinoside in human plasma and cerebrospinal fluid is described. Complete separation from endogenous constituents was achieved by isocratic reversed-phase chromatography using phosphate buffer (0.05 M, pH 7.0) as the eluent. The limit of detection was 50 ng/ml. Day-to-day coefficients of variation were below 10%. The applicability of this rapid, simple and specific method for pharmacokinetic studies and monitoring of therapy was demonstrated.

Corticosteroid response in chronic lung disease of prematurity.

The medical records of 23 infants with stable chronic lung diseases of prematurity who were treated with corticosteroids during the second month of life were reviewed. Thirteen (56.5%) had significant improvement in lung function as evidenced by a mean decrease in delta AaPO2 of 49.5% and a mean decrease in PaCO2 of 15.4%. These proved to be long-lasting effects. Infants who responded to corticosteroids had significantly lower mean gestational age, birthweight, and percent loss of birthweight when compared with the infants who did not improve with this treatment. In addition, the number of days they required supplemental oxygen were fewer. Thus, corticosteroids may be beneficial therapeutically for some premature infants with chronic lung disease. Alternatively, corticosteroid responsiveness may differentiate types of chronic lung disease with improved prognoses. Controlled clinical trials are necessary before corticosteroids can be recommended in treating or evaluating chronic lung disease of prematurity.