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.

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.

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.

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.

[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.

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.

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.

Comparison of three methods for obtaining principal components from family data in genetic analysis of complex disease.

Three multivariate techniques used to derive principal components (PCs) from family data were compared for their ability to model family data and power to detect linkage. Using the simulated data from Genetic Analysis Workshop 12, the five quantitative traits were first adjusted for age, sex, and environmental factors 1 and 2. Then, standard PCs, PCs obtained from between-family covariance, and PCs obtained from within-family genetic covariance were derived and subjected to multivariate sib pair linkage analysis. The standard PCs obtained from the overall correlation matrix allowed identification of key features of the true genetic model more readily than did the other methods. For detection of linkage, standard PCs and PCs obtained from the between-family genetic covariance performed similarly in terms of both power and type 1 error, and both methods performed better than the PCs obtained from within-family genetic covariance.

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.

The effect of multiple neuroimaging studies on classification, treatment, and outcome of acute ischemic stroke.

OBJECTIVE: To examine the effect of serial neuroimaging studies on the diagnosis, therapy, and outcome of patients with acute stroke. DESIGN: Retrospective case series. SETTING: Tertiary care teaching hospital. PATIENTS: 206 adult patients (mean age +/- SD, 66.0 +/- 10.8 years) hospitalized with a diagnosis of acute stroke between 1990 and 1993. MEASUREMENTS: Strokes were retrospectively assigned to five categories (large-vessel, small-vessel, cardioembolic, other, or unknown) using standardized criteria based on the history, physical examination, ancillary test results, and first computed tomographic (CT) or magnetic resonance imaging (MRI) study of the head. Strokes were reclassified after the results of further neuroimaging studies, if any, were reviewed. The type and timing of therapy and the patient outcome at hospital discharge were documented. RESULTS: The additional studies changed stroke classification in only 20.0% of the 140 patients who had two or more neuroimaging studies. All classification changes were from the unknown cause category to a category with a specific cause. In most patients receiving treatment (93.2%), therapy began before an additional CT or MRI study was obtained. In patients who had one neuroimaging study, 70.1% went home, 24.0% went to a skilled nursing facility, and 5.9% died; the corresponding percentages in persons who had multiple studies were 73.3%, 24.4%, and 2.2% (P > 0.1). CONCLUSIONS: Serial neuroimaging studies did not alter the classification of strokes for which an initial diagnosis had already been made. However, they were useful in determining the cause of strokes initially classified as having an unknown cause. Therapy was almost always begun immediately after the first CT or MRI study was obtained. Outcome at hospital discharge was not significantly related to the number of neuroimaging studies obtained.

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.

Molecular cloning and DNA sequence analysis of pepL, a leucyl aminopeptidase gene from Lactobacillus delbrueckii subsp. lactis DSM7290.

A genomic library of Lactobacillus delbrueckii subsp. lactis DSM7290 DNA fragments from a Sau3A partial digestion in the low-copy-number vector pLG339, was used to screen Escherichia coli for the presence of peptidases. Using the chromogenic substrate leucine-beta-naphthylamide (Leu-NH-Nap) and E. coli strain CM89 lacking the corresponding enzyme activity in an enzymic plate assay, allowed the isolation of two peptidase genes; the newly described pepL and the recently cloned and sequenced pepN. Clones could be distinguished not only by the restriction pattern of isolated plasmids but also by the rate and intensity of their colour reaction with Leu-NH-Nap. Three out of five clones were identified to express the Lactobacillus pepN gene; the others were shown to express a second aminopeptidase gene, designated pepL. This gene, together with 200 bp upstream of the proposed AUG initiation codon, was further subcloned and sequenced. The corresponding open reading frame of 897 nucleotides is predicted to encode a protein of 299 amino acids (34,541 Da). Searching the EMBL database revealed similarity to the prolinase of Lactobacillus helveticus (45.8% identity), to the iminopeptidases of Lb. delbrueckii subsp. lactis and Lb. delbrueckii subsp. bulgaricus (25.5%), and to the Bacillus coagulans prolinase (21.5%). Minor similarities were detected for hydrolytic enzymes with serine active sites. The product encoded by the pepL gene was functional but could not be visualized on Coomassie-blue-stained polyacrylamide gels. High level expression of peptidase L in E. coli was achieved by placing the gene under the control of the T7 promoter.

Enhancement of neonatal neutrophil motility (chemotaxis) with adult fresh frozen plasma.

We examined the effect of adult fresh frozen plasma (FFP) on neonatal neutrophil (PMN) motility (chemotaxis) using a micropore filter assay. Adult FFP was transfused into 13 neonates receiving FFP transfusion for suspected life-threatening sepsis. Blood was obtained from neonates before and after FFP transfusion for assessment of PMN chemotaxis. An increase in PMN chemotaxis was noted in 12 of the 13 neonates following FFP transfusion, with a mean percentage increase of 12 +/- 3% (p less than 0.01). PMN chemotaxis increased 13 +/- 2% (p less than 0.01) in four bacteremic infants and 11 +/- 5% (p = 0.06) in nine infants without bacteremia. Adult FFP transfusion may enhance impaired neonatal PMN motility and improve outcome from infection in newborn infants.

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.

Two distinct adaptive responses in the synthesis of exocrine pancreatic enzymes to inverse changes in protein and carbohydrate in the diet.

Rates of synthesis of 16 individual pancreatic exocrine proteins; tissue concentrations of amylase, trypsinogen, and chymotrypsinogen; and morphological assessment of pancreatic acinar cells were studied in the exocrine pancreas in response to inverse changes in protein and carbohydrate in the diet, administered for 12 days. Two distinct patterns of response were observed. During adaptation to diets containing normal protein (22%) or increased levels of protein (30, 45, 64, and 82% protein) and correspondingly decreased levels of carbohydrate, amylase and the majority of protease zymogens were synthesized in direct proportion to the nutritional substrates carbohydrate and protein, respectively, in the diet. With increases in dietary protein, anticoordinate patterns of response in the synthesis of exocrine isoenzymes were observed: 0.4- to 2.0-fold increases in trypsinogen forms 1 and 2, chymotrypsinogen forms 1 and 2, proelastase 1, and procarboxypeptidases A and B; 5- to 7-fold decreases in amylase forms 1 and 2; and insignificant changes in trypsinogen 3, proelastase 2, lipase, and ribonuclease. During adaptation to diets containing normal protein (22%) or decreased levels of protein (0 or 10% protein) and correspondingly increased levels of carbohydrate, amylase and the majority of protease zymogens were synthesized in inverse proportion to nutritional substrates in the diet.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

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.