SARS-CoV-2 Distribution in Residential Housing Suggests Contact Deposition and Correlates with Rothia sp.

Monitoring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on surfaces is emerging as an important tool for identifying past exposure to individuals shedding viral RNA. Our past work demonstrated that SARS-CoV-2 reverse transcription-quantitative PCR (RT-qPCR) signals from surfaces can identify when infected individuals have touched surfaces and when they have been present in hospital rooms or schools. However, the sensitivity and specificity of surface sampling as a method for detecting the presence of a SARS-CoV-2 positive individual, as well as guidance about where to sample, has not been established. To address these questions and to test whether our past observations linking SARS-CoV-2 abundance to Rothia sp. in hospitals also hold in a residential setting, we performed a detailed spatial sampling of three isolation housing units, assessing each sample for SARS-CoV-2 abundance by RT-qPCR, linking the results to 16S rRNA gene amplicon sequences (to assess the bacterial community at each location), and to the Cq value of the contemporaneous clinical test. Our results showed that the highest SARS-CoV-2 load in this setting is on touched surfaces, such as light switches and faucets, but a detectable signal was present in many untouched surfaces (e.g., floors) that may be more relevant in settings, such as schools where mask-wearing is enforced. As in past studies, the bacterial community predicts which samples are positive for SARS-CoV-2, with Rothia sp. showing a positive association. IMPORTANCE Surface sampling for detecting SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19), is increasingly being used to locate infected individuals. We tested which indoor surfaces had high versus low viral loads by collecting 381 samples from three residential units where infected individuals resided, and interpreted the results in terms of whether SARS-CoV-2 was likely transmitted directly (e.g., touching a light switch) or indirectly (e.g., by droplets or aerosols settling). We found the highest loads where the subject touched the surface directly, although enough virus was detected on indirectly contacted surfaces to make such locations useful for sampling (e.g., in schools, where students did not touch the light switches and also wore masks such that they had no opportunity to touch their face and then the object). We also documented links between the bacteria present in a sample and the SARS-CoV-2 virus, consistent with earlier studies.

SARS-CoV-2 Distribution in Residential Housing Suggests Contact Deposition and Correlates with Rothia sp.

Monitoring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on surfaces is emerging as an important tool for identifying past exposure to individuals shedding viral RNA. Our past work has demonstrated that SARS-CoV-2 reverse transcription-quantitative PCR (RT-qPCR) signals from surfaces can identify when infected individuals have touched surfaces such as Halloween candy, and when they have been present in hospital rooms or schools. However, the sensitivity and specificity of surface sampling as a method for detecting the presence of a SARS-CoV-2 positive individual, as well as guidance about where to sample, has not been established. To address these questions, and to test whether our past observations linking SARS-CoV-2 abundance to Rothia spp. in hospitals also hold in a residential setting, we performed detailed spatial sampling of three isolation housing units, assessing each sample for SARS-CoV-2 abundance by RT-qPCR, linking the results to 16S rRNA gene amplicon sequences to assess the bacterial community at each location and to the Cq value of the contemporaneous clinical test. Our results show that the highest SARS-CoV-2 load in this setting is on touched surfaces such as light switches and faucets, but detectable signal is present in many non-touched surfaces that may be more relevant in settings such as schools where mask wearing is enforced. As in past studies, the bacterial community predicts which samples are positive for SARS-CoV-2, with Rothia sp. showing a positive association. IMPORTANCE: Surface sampling for detecting SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19), is increasingly being used to locate infected individuals. We tested which indoor surfaces had high versus low viral loads by collecting 381 samples from three residential units where infected individuals resided, and interpreted the results in terms of whether SARS-CoV-2 was likely transmitted directly (e.g. touching a light switch) or indirectly (e.g. by droplets or aerosols settling). We found highest loads where the subject touched the surface directly, although enough virus was detected on indirectly contacted surfaces to make such locations useful for sampling (e.g. in schools, where students do not touch the light switches and also wear masks so they have no opportunity to touch their face and then the object). We also documented links between the bacteria present in a sample and the SARS-CoV-2 virus, consistent with earlier studies.

Analysis of SARS-CoV-2 RNA Persistence across Indoor Surface Materials Reveals Best Practices for Environmental Monitoring Programs.

Environmental monitoring in public spaces can be used to identify surfaces contaminated by persons with coronavirus disease 2019 (COVID-19) and inform appropriate infection mitigation responses. Research groups have reported detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on surfaces days or weeks after the virus has been deposited, making it difficult to estimate when an infected individual may have shed virus onto a SARS-CoV-2-positive surface, which in turn complicates the process of establishing effective quarantine measures. In this study, we determined that reverse transcription-quantitative PCR (RT-qPCR) detection of viral RNA from heat-inactivated particles experiences minimal decay over 7 days of monitoring on eight out of nine surfaces tested. The properties of the studied surfaces result in RT-qPCR signatures that can be segregated into two material categories, rough and smooth, where smooth surfaces have a lower limit of detection. RT-qPCR signal intensity (average quantification cycle [Cq]) can be correlated with surface viral load using only one linear regression model per material category. The same experiment was performed with untreated viral particles on one surface from each category, with essentially identical results. The stability of RT-qPCR viral signal demonstrates the need to clean monitored surfaces after sampling to establish temporal resolution. Additionally, these findings can be used to minimize the number of materials and time points tested and allow for the use of heat-inactivated viral particles when optimizing environmental monitoring methods. IMPORTANCE Environmental monitoring is an important tool for public health surveillance, particularly in settings with low rates of diagnostic testing. Time between sampling public environments, such as hospitals or schools, and notifying stakeholders of the results should be minimal, allowing decisions to be made toward containing outbreaks of coronavirus disease 2019 (COVID-19). The Safer At School Early Alert program (SASEA) (https://saseasystem.org/), a large-scale environmental monitoring effort in elementary school and child care settings, has processed >13,000 surface samples for SARS-CoV-2, detecting viral signals from 574 samples. However, consecutive detection events necessitated the present study to establish appropriate response practices around persistent viral signals on classroom surfaces. Other research groups and clinical labs developing environmental monitoring methods may need to establish their own correlation between RT-qPCR results and viral load, but this work provides evidence justifying simplified experimental designs, like reduced testing materials and the use of heat-inactivated viral particles.

Comparison of heat-inactivated and infectious SARS-CoV-2 across indoor surface materials shows comparable RT-qPCR viral signal intensity and persistence.

Environmental monitoring in public spaces can be used to identify surfaces contaminated by persons with COVID-19 and inform appropriate infection mitigation responses. Research groups have reported detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) on surfaces days or weeks after the virus has been deposited, making it difficult to estimate when an infected individual may have shed virus onto a SARS-CoV-2 positive surface, which in turn complicates the process of establishing effective quarantine measures. In this study, we determined that reverse transcription-quantitative polymerase chain reaction (RT-qPCR) detection of viral RNA from heat-inactivated particles experiences minimal decay over seven days of monitoring on eight out of nine surfaces tested. The properties of the studied surfaces result in RT-qPCR signatures that can be segregated into two material categories, rough and smooth, where smooth surfaces have a lower limit of detection. RT-qPCR signal intensity (average quantification cycle (Cq)) can be correlated to surface viral load using only one linear regression model per material category. The same experiment was performed with infectious viral particles on one surface from each category, with essentially identical results. The stability of RT-qPCR viral signal demonstrates the need to clean monitored surfaces after sampling to establish temporal resolution. Additionally, these findings can be used to minimize the number of materials and time points tested and allow for the use of heat-inactivated viral particles when optimizing environmental monitoring methods.

Matrices for cartilage repair.

Techniques for repairing focal articular cartilage defects are evolving from methods that induce a local stimulation of fibrocartilaginous repair to methods that will lead to a hyaline articular cartilage repair. Mosaicplasty and autologous chondrocyte implantation are examples of the latter. A tissue engineered hyaline cartilage implant that could be used off the self would minimize the morbidity of these techniques. However, there are significant questions that still need to be resolved before such tissue-engineered implants will be practical. Principally among these is the question of what is the ideal matrix for such an implant, particularly from the standpoint of the best material and architecture. Second, what is the ideal cell source to use with these implants. A third major unknown is what is the most ideal way to use growth factors to enhance the repair. As these issues are resolved, the prospects of a tissue engineered cartilage replacement will advance from theory to practice.

Donor cell fate in tissue engineering for articular cartilage repair.

Articular cartilage repair is a clinical challenge because of its limited intrinsic healing potential. Considerable research has focused on tissue engineering and transplantation of viable chondrogenic cells to enhance cartilage regeneration. However, the question remains: do transplanted allogenic cells survive in the repair with time? This study assessed donor cell fate after transplantation of male New Zealand White rabbit perichondrium cell and polylactic acid constructs into osteochondral defects created in the medial femoral condyles of female New Zealand White rabbits. Repair tissue was harvested at 0, 1, 2, 3, 7, and 28 days after implantation and was evaluated for cell viability and total cell number using confocal microscopic analysis. The number of donor cells in each sample was estimated using quantitative polymerase chain reaction targeting a gender-specific gene present on the Y-chromosome, the sex-determining region Y gene, and a control deoxyribonucleic acid present in male and female cell deoxyribonucleic acid, the matrix metalloproteinase-1 gene promoter. Average cell viability was found to be 87% or more at all times. Donor cells were present in repair tissue for 28 days after implantation. However, the number of donor cells declined from approximately 1 million at Time 0 to approximately 140,000 at 28 days. This decline in donor cells was accompanied by a significant influx of host cells into the repair tissue. This study shows that the sex-determining region Y gene is a valuable marker for tracking the fate of transplanted allogenic cells in tissue engineering.

Multimethod assessment of psychopathology among DSM-IV subtypes of children with attention-deficit/hyperactivity disorder: self-, parent, and teacher reports.

Using data based on self-, parent, and teacher reports, we assessed various aspects of psychopathology in a large sample of control children and those with ADHD. Confirmatory factor analysis was employed to extract response bias from latent constructs of aggression, anxiety, attention problems, depression, conduct disorder, and hyperactivity. These latent constructs were then entered into logistic regression equations to predict membership in control versus ADHD groups, and to discriminate between ADHD subtypes. Results of the regression equations showed that higher levels of attention problems and aggression were the best predictors of membership in the ADHD group relative to controls. Logistic regression also indicated that a higher degree of aggression was the only significant predictor of membership in the ADHD-Combined group compared to the ADHD-Inattentive group. However, when comorbid diagnoses of Oppositional Defiant Disorder and Conduct Disorder were controlled for in the logistic regression, greater hyperactivity rather than aggression was the sole variable with which to distinguish the ADHD-Combined from the ADHD-Inattentive subtype. Results are discussed in the context of the DSM-IV ADHD nosology and the role of instrument and source bias in the diagnosis of ADHD.

PT-12, a putative ras-activated proliferation-dependent gene, is expressed in patellar tendon and not in anterior cruciate ligament.

We describe a gene (PT-12) that is expressed in the patellar tendon and not in the anterior cruciate ligament. We used a recently developed polymerase chain reaction-based subtractive cDNA analysis to discover genes that are overexpressed in the patellar tendon but not expressed in the anterior cruciate ligament; the long-term objective was to find genes that are central to the self-repair of the patellar tendon, in contrast with the inability of the anterior cruciate ligament to launch a repair response following injury. PT-12 is a homologue of human S2 or mouse LLRep3 ribosomal genes, which are known to be overexpressed in highly proliferating cells. This study opens a new vista to the development of techniques and reagents to study the differences between two periarticular tissues (i.e., the patellar tendon and anterior cruciate ligament) that differ primarily in their ability to self-repair.

Convergent and criterion-related validity of the Behavior Assessment System for Children-Parent Rating Scale.

Examined aspects of the validity of the Behavior Assessment System for Children (BASC; Reynolds & Kamphaus, 1992) Parent Rating Scale (BASC-PRS) in 156 children with cross-setting disruptive behavior. The BASC-PRS is a recently published multidimensional measure composed of conceptually derived scales created for use in conjunction with psychiatric and educational classification systems. Convergent validity was assessed through correlations of BASC-PRS scale scores with scale scores on the Child Behavior Checklist/4-18 (CBCL/4-18; Achenbach, 1991b). Criterion-related validity was evaluated as the ability of BASC-PRS and CBCL/4-18 scales to predict membership in diagnostic groupings (no diagnosis, attention deficit hyperactivity disorder [ADHD] only, and ADHD with a comorbid externalizing disorder) derived via structured interviews based on the third, revised edition of the Diagnostic and Statistical Manual of Mental Disorders (American Psychiatric Association, 1987). Results showed the validity of the BASC-PRS to be comparable to that of the CBCL/4-18. Given its conceptually derived scales, the BASC-PRS may prove to be a useful-tool for assessing childhood disruptive behavior.

Specific competitive inhibitor of secreted phospholipase A2 from berries of Schinus terebinthifolius.

Two structurally related triterpenoids 1 and 2 from pink peppercorn (berries of Schinus terebinthifolius) are identified and characterized as active site-directed specific competitive inhibitors of the three classes of secreted 14 kDa phospholipase A2. The inhibitors not only protect the active site histidine from alkylation but also inhibit the action of secreted phospholipase A2 from pig pancreas, human synovial fluid, and bee venom. Detailed X-ray crystallographic results on the structures of the inhibitors are provided. By physical methods and X-ray crystallography the triterpenoids were identified as masticadienoic acid and masticadienolic acid (schinol). Several other triterpenoids were ineffective as inhibitors of phospholipase A2; however certain ganoderic acid derivatives showed noticeable inhibition. Results show that the side chain of these acidic tetracyclic terpenoids can access the catalytic-site region of phospholipase A2, whereas the acyclic nucleus is at the interfacial recognition region. The selectivity of the assay protocol used here is demonstrated by the fact that the original screen of ethyl acetate extracts of 60 commercially available spices and herbs was carried out with phospholipase A2 from pig pancreas, and only one extract showed inhibitory action on the hydrolytic activity in the scooting mode. Under such assay conditions the enzyme remains tightly bound to the surface of the substrate vesicles. In this way, nonspecific effects of additives that promote desorption of the enzyme from the substrate vesicle surface, under conditions in which the binding of the enzyme to the vesicle is weak, are avoided. The assay protocol is useful for the kinetic characterization of the inhibitors of phospholipase A2, and it does not give false positive results with amphiphilic and hydrophobic compounds, as is the case with virtually all assay systems in use.

Developmental and symptom specificity of hopelessness, cognitive errors, and attributional bias among clinic-referred youth.

The present study examined the unique and interactive relationships between age and indices of psychopathology (i.e., anxiety, aggression, and depression), with three types of maladaptive cognitions: hopelessness, negative cognitive errors, and attributional bias. Some negative cognitions were not unique to depression and were associated with broader psychopathology. Developmental considerations also influenced some negative cognitions or qualified the association between negative cognitions and depression.

Structure of a dihydrooxazole oxa-bridged octalin.

C26H41NO7, Mr = 479.61, triclinic, P1, a = 11.894 (4), b = 12.360 (2), c = 9.969 (2) A, alpha = 106.11 (2), beta = 100.68 (2), gamma = 99.02 (2) degrees, V = 1349.5 (6) A3, Z = 2, Dx = 1.180 Mg m-3, Mo K alpha, lambda = 0.71069 A, mu = 0.079 mm-1, F(000) = 520, T = 296 K, R = 0.041, wR = 0.047 for 3196 observed unique reflections. A new approach to the total synthesis of the nargenicin macrolide system employing a [2,3] Wittig rearrangement to control the remote C16-C17 stereochemistry required characterization of a pivotal intermediate by X-ray single-crystal methods. The structure is characterized by the formation of strong intermolecular alcohol to dihydrooxazole-N hydrogen bonds, an essentially strain-free tricyclic oxa-bridged octalin nucleus, and low-energy conformations of the methoxymethyl ether side chains.

Effects of a school-based cognitive-behavioral intervention for ADHD children.

Two variations of school-based cognitive-behavioral training (CBT) program were compared to each other and to a waiting-list control condition in the treatment of children with attention-deficit hyperactivity disorder (ADHD). The experimental interventions included a multicomponent condition that provided coordinated training programs for parents, teachers, and children and a teacher-only condition that offered training for classroom teachers only. Evaluation of outcome occurred at pre-intervention, post-intervention and at 6-week followup periods. Depedent measures included classroom behavior observations, teacher ratings of child behavior, child self-report, and teacher ratings of adjustment. The multicomponent CBT condition was significantly better than the other conditions at improving observed off-task/disruptive behavior at post-test. This improvement was maintained at followup, although treatment condition differences were no longer significant. There were no treatment condition differences on any other measures at post-intervention or followup. It was concluded that the intervention had minimal short-term effects on the ADHD children. The results are discussed within the context of several methodological limitations of the study which serve as proposals for continued research in this area.

The direct observation of a psoralen-thymine UVA induced solid-state cycloaddition reaction product by single-crystal x-ray diffractometry.

Single-crystal x-ray diffraction methods have been used to directly observe and simultaneously determine the molecular structure of the UVA induced cis-syn photocycloaddition product in a partially photolyzed single crystal of a psoralen(pyrone ring side)-DNA(thymine) interaction model compound, 1'-(8-oxypsoralen)-8'(thym-1"yl)3',6'-dioxaoctane.