Heparin-mediated PCR interference in SARS-CoV-2 assays and subsequent reversal with heparinase I.

Heparin is postulated to block the interaction of SARS-CoV-2 with highly glycosylated proteins which are critical for binding the angiotensin-converting enzyme 2 (ACE2), an essential mechanism for host-cell entry and viral replication. Intranasal heparin is under investigation for use as a SARS-CoV-2 preventative in the IntraNasal Heparin Trial (INHERIT, NCT05204550). Heparin directly interferes with real-time quantitative polymerase chain reaction (RT-qPCR), the gold standard for SARS-CoV-2 detection. This study aimed to investigate the magnitude of heparin interference across various clinical laboratory testing platforms, and the reversal of any interference by degradation of heparin using the heparinase I enzyme in nasopharyngeal swab (NP) samples for SARS-CoV-2 analysis by RT-qPCR. Heparin-mediated PCR interference was evident at heparin concentrations as low as 10 IU/mL across all platforms tested, with the exclusion of the Hologic Panther Aptima SARS-CoV-2 assay. Rates of false negative or invalid results increased with increasing heparin concentrations on all platforms, except the Hologic Panther Aptima and Roche Cobas LIAT. Heparinase I reversed heparin-mediated PCR inhibition across in all samples tested, except those with initial Ct values >35. Our study shows that the use of heparin-containing nasal sprays interferes with the detection of SARS-CoV-2 in NP swab samples by RT-qPCR, a phenomenon that is not well recognised in the literature. Furthermore, this study has also demonstrated that heparin-mediated PCR inhibition can be prevented through heparinase I treatment, demonstrating restoration of clinically significant results with Ct values <35.

A placental model of SARS-CoV-2 infection reveals ACE2-dependent susceptibility and differentiation impairment in syncytiotrophoblasts.

SARS-CoV-2 infection causes COVID-19. Several clinical reports have linked COVID-19 during pregnancy to negative birth outcomes and placentitis. However, the pathophysiological mechanisms underpinning SARS-CoV-2 infection during placentation and early pregnancy are not clear. Here, to shed light on this, we used induced trophoblast stem cells to generate an in vitro early placenta infection model. We identified that syncytiotrophoblasts could be infected through angiotensin-converting enzyme 2 (ACE2). Using a co-culture model of vertical transmission, we confirmed the ability of the virus to infect syncytiotrophoblasts through a previous endometrial cell infection. We further demonstrated transcriptional changes in infected syncytiotrophoblasts that led to impairment of cellular processes, reduced secretion of HCG hormone and morphological changes vital for syncytiotrophoblast function. Furthermore, different antibody strategies and antiviral drugs restore these impairments. In summary, we have established a scalable and tractable platform to study early placental cell types and highlighted its use in studying strategies to protect the placenta.

Federal prison residential drug treatment reduces substance use and arrests after release.

OBJECTIVE: The effectiveness of federal prison-based residential drug and alcohol treatment programs was evaluated using event history procedures that addressed the problem of selection bias and included a wide range of control variables. METHODS: The sample comprised 760 treatment subjects and 809 comparison subjects. Treatment subjects were from 20 different prisons of medium, low, and minimum security levels. Comparison subjects were drawn from over 30 prisons. RESULTS: The results indicated that individuals who entered and completed in-prison residential treatment were less likely to experience the critical postrelease outcomes of new arrests and substance use during the first 6 months following release. CONCLUSIONS: Without controlling for selection bias, the effects of treatment would most likely have been attenuated. The results have greater generalizability than other studies of prison-based treatment. This study occurred within a multisite context of 20 programs serving both male and female inmates and operating within different security levels and different geographic regions.

The stigma scale: measuring body image and the skin.

People with chronic wounds or skin conditions often face not only physiological challenges, but also emotional ones as well. Measuring these emotional challenges and, more broadly, the emotional challenges of having a skin defect was the focus of this instrument development study.

Are ethics committee members competent to consult?

The authors describe research conducted to assess the skills and knowledge base of individuals who perform ethics consultations in Maryland hospitals. The findings indicate a lack of formal educational preparation on the part of those who perform consults, and a general lack of institutional support for ethics committees.

Living with a chronic wound: the voices of sufferers.

As the American population continues to age and people live longer, a growing number of individuals have chronic health problems. Of these health problems, chronic wounds are among the costliest. The focus of this research was to understand the meaning of having a nonhealing wound. Heideggerian hermeneutics--one of several approaches to interpretive scholarship--was used in this study. Through interpreting language or texts, Heideggerian hermeneutics seeks to reveal meanings. In this study, the researchers interviewed 10 participants with chronic wounds and performed data analysis of the transcripts using qualitative software, and then Diekelmann's framework of Heideggerian hermeneutical analysis. Two constitutive patterns with six themes emerged from the data--"contending with the wound" and "staying home; staying back." Profound themes of oozing and smelling, losing sleep, being in pain, and noticing are described in the participants own words in the text. By listening to the participants' stories and understanding what it is like to live with a chronic wound, healthcare providers can create caring contexts.

Antisocial personality and depression among incarcerated drug treatment participants.

PURPOSE: The co-occurrence of antisocial personality (ASP) and depression among drug-dependent individuals was examined in a sample of federal inmates participating in residential or outpatient drug and alcohol treatment to increase our understanding of the co-occurrence of these disorders. METHODS: We examined drug dependence patterns both by the number of drugs of dependence as well as by type of drug, and separately studied men and women. The Diagnostic Interview Schedule (DIS) was used to obtain DSM-III-R diagnostic information on a sample of 609 men and women participating in a multi-site drug treatment evaluation project. Logistic regression results are reported that compared lifetime rates of ASP and major depression by number of drugs of dependency for men and women. ASP and major depression diagnostic rates were also examined by type of drug dependency pattern among men. RESULTS: We found that both the number of drugs as well as the type of drug(s) are related to prevalence patterns for both diagnoses. The high rates of ASP and major depression among specific subgroups of drug-dependent inmates highlight the need for thorough psychiatric assessment and the tailoring of treatment programs to the issues associated with these diagnoses. IMPLICATIONS: Our results suggest that although there are similarities in co-morbidity between men and women, the differences point to the need to study men and women separately.

The Stigma Scale: measuring body image and the skin.

People with chronic wounds or skin conditions often face not only physiological challenges, but also emotional ones as well. Measuring these emotional challenges and more broadly, the emotional challenges of having a skin defect, was the focus of this instrument development study.

Morphometric comparison of Chinese-origin and Indian-derived rhesus monkeys (Macaca mulatta).

Chinese-origin and Indian-derived rhesus monkeys (Macaca mulatta), ranging in age from 6 months to 15 years, were weighed and measured during the 1995 inventory of the SAIDS breeding colony at the Tulane Regional Primate Research Center. Data were analyzed separately for males (n=279) and females (n=554), and an analysis of variance was done for five measurements and two indices, with age and country of origin as independent variables. All measurements increased significantly with age, but Chinese-origin males were heavier, longer, and taller than Indian-derived males. Chinese-origin juvenile females were heavier and longer than Indian-derived juvenile females, but this pattern was reversed for adults. Chinese-origin rhesus monkeys exhibited more adult sexual dimorphism than did Indian-derived rhesus monkeys. An analysis of Chinese-origin adults that were Louisiana-born vs. China-born indicated that Louisiana-born adults were lighter, longer, and taller than their China-born parental generation. These differences were statistically significant for adult females and descriptively suggestive for adult males, and both suggest body and appendage adaptation to a warmer, wetter climate.

Transition theory and its relevance to patients with chronic wounds.

A wound, in the broadest sense, is a disruption of normal anatomic structure and function. Acute wounds progress through a timely and orderly sequence of repair that leads to the restoration of functional integrity. In chronic wounds, this timely and orderly sequence goes awry. As a result, people with chronic wounds often face not only physiological difficulties but emotional ones as well. The study of body image and its damage as a result of a chronic wound fits well with Selder's transition theory. This article describes interviews with seven patients with chronic wounds. The themes that emerged from those interviews were compared with Selder's theory to describe patients' experience with chronic wounds as a transition process that can be identified and better understood by healthcare providers.

A comparison of two culturing methods for chronic wounds.

Bacterial infection has always been a potential complication of any wound. Controversy exists regarding the significance of bacteria in chronic wounds. It is important to accurately diagnose wound infection by bacterial identification and quantification in order to prevent unnecessary and/or inappropriate treatments and to minimize patient complications. The primary function of culturing is to identify infection in a wound. The tissue culture is an accepted standard for measuring infection, although swab cultures are commonplace in the clinical setting. The purpose of this study was to test the differences in bacterial counts and identification in swab and tissue cultures taken from the same wound site of 10 chronic wounds. It was hypothesized that if swab and tissue cultures are equally effective in identifying and quantifying the organisms in a chronic wound, they are equally effective methods in determining infection in the chronic wound. The reliability, validity and limitations of the study are discussed, as well as the statistical analyses and results.

Oxidation of LDL: role in atherogenesis.

Oxidation of LDL is proposed to accelerate atherogenesis by the following sequence of events. LDL accumulates in atherosclerotic plaques, presumably due to interaction with intimal proteoglycans. The LDL then undergoes oxidation, and aldehydic products of lipid peroxidation such as HNE or other aldehyde products derived from lipid peroxidation, induce blocking of lysine residues on apo B. This results in its recognition by the scavenger receptor on tissue macrophages at sites in which LDL concentrations are low. At sites in which the LDL concentration is high, modification with such products induces intermolecular cross-linking and particle aggregation. The aggregated, oxidized LDL particles are then phagocytosed by tissue macrophages to induce lipid loading of these cells and the formation of foam cells, a characteristic of the earliest atherosclerotic lesion. By these mechanisms oxidation of LDL accelerates atherogenesis.

Teaching basic ethical concepts and decision-making: a staff development application.

Academic nursing programs are increasing the amount of ethical instruction presented to undergraduate and graduate students. However, nurses who graduated several years ago may find themselves without the background and skills needed to discuss the complex bioethical dilemmas they now face. This article presents the development of a continuing education program to acquaint nurses with basic ethical concepts and bioethical decision-making processes. Presentations were piloted with a small group of nurses from the outpatient department of a pediatric tertiary care hospital.

Quantification of apo[a] and apoB in human atherosclerotic lesions.

Lipoprotein[a] or Lp[a] is a cholesterol-rich plasma lipoprotein that is associated with increased risk for cardiovascular disease. To better understand this association we determined the amount of apo[a] and apoB as possible estimates for Lp[a] and low density lipoprotein (LDL) accumulation in atherosclerotic lesions and in plasma, from patients undergoing vascular surgery, using specific radioimmunoassays for apolipoprotein[a] and apolipoprotein B. Apo[a] and apoB were operationally divided into a loosely bound fraction obtained by extracting minced samples of plaque with phosphate-buffered saline (PBS), and a tightly bound fraction obtained by extracting the residual tissue with 6 M guanidine-HCl (GuHCl). We found that 83% of all apo[a] but only 32% of all apoB in lesions was in the tightly bound fraction. When normalized for corresponding plasma levels, apo[a] accumulation in plaques was more than twice that of apoB. All fractions of tissue apo[a], loosely bound, tightly bound, and total, correlated significantly with plasma apo[a]. However, no significant correlations were found between any of the tissue fractions and plasma apoB. If all apo[a] and apoB had been associated with intact Lp[a] or LDL particles, the calculated mass of tightly bound Lp[a] would actually have exceeded that of tightly bound LDL in five cases with plasma Lp[a] levels above 5 mg apo[a] protein/dl. When PBS and GuHCl extracts of lesions were subjected to one-dimensional electrophoresis, the major band stained for lipid and immunoblotted positively for apo[a] and apoB, suggesting the presence of some intact Lp[a] in these extracts. These results suggest that Lp[a] accumulates preferentially to LDL in plaques, and that plaque apo[a] is directly associated with plasma apo[a] levels and is in a form that is less easily removable than most of the apoB. This preferential accumulation of apo[a] as a tightly bound fraction in lesions, could be responsible for the independent association of Lp[a] with cardiovascular disease in humans.

Mycotic embolism and embolomycotic aneurysms. Neglected lessons of the past.

During the past decade, nine patients with bacterial endocarditis have required management of mycotic emboli and/or aneurysms in this center. In these patients, 25 separate mycotic emboli or aneurysms were identified. Among these were four visceral, 11 lower extremity, one aortic, one hypogastric, and eight cerebral lesions. Multiple sites were involved in seven of the nine patients (78%). Presenting symptoms were secondary to acute expansion of mycotic aneurysms in three patients and secondary to rupture of aneurysms in four patients. Mycotic emboli produced cerebral infarction in two patients and acute ischemia in six patients. Asymptomatic mycotic aneurysms of the middle cerebral, hepatic, hypogastric, and profunda femoris arteries and asymptomatic emboli to the profunda femoris and tibial arteries were found during angiographic study. Management included resection alone (7 aneurysms), resection and graft replacement (2 aneurysms and 2 emboli), embolectomy (2), or observation. There was no mortality or loss of limb in these patients. This experience underscores the frequent multiplicity of mycotic emboli and/or aneurysms and stresses the importance of empiric angiographic survey to exclude silent yet potentially lethal visceral and cerebral mycotic foci in patients with bacterial endocarditis and peripheral emboli or aneurysms.