Coronavirus Infections in Animals: Risks of Direct and Reverse Zoonoses.

The publications on animal coronavirus infections that have the greatest emerging potential, as well as official data from the World Organization for Animal Health (OIE) on cases of animal infection with COVID-19, are analyzed. Like most infectious diseases common to humans, coronavirus infections were first discovered in animals. Due to the increased rate of replication and recombination activity compared to other viruses, mutations occur more often in the genome of coronaviruses, which contribute to the acquisition of new qualities in order to consolidate in the host organism. Examples of cross-species transmission are not only SARS-CoV, MERS-CoV, and SARS-CoV-2, which are dangerous to humans, but also coronaviruses of agricultural and domestic animals, between which there is a genetic relationship. There are several known cases of zoo, wild, domestic, and farm animals displaying symptoms characteristic of COVID-19 and identification of the genome of the SARS-CoV-2 virus in them. The issue of cross-species transmission of coronavirus infections, in particular the reverse zoonosis of SARS-CoV-2 from animals to humans, is widely discussed. According to the conclusions of many researchers, including OIE experts, there is no direct evidence base for infection of humans with COVID-19 from animals. However, people with suspected COVID-19 and with a confirmed diagnosis are still advised to isolate not only from people but also from animals. A number of methods for specific prevention, diagnosis, and immunization against a wide range of coronavirus infections are being developed at the All-Russia Research Institute for Animal Protection.

Persistent organic pollutants in plastic marine debris found on beaches in San Diego, California.

Plastic debris were collected from eight beaches around San Diego County, California. Debris collected include: pre-production pellets and post-consumer plastics including fragments, polystyrene (PS) foam, and rubber. A total of n = 2453 pieces were collected ranging from <5 mm to 50 mm in size. The plastic pieces were separated by type, location, and appearance and analyzed for polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its breakdown products, and chlordanes. PAH concentrations ranged from 30 ng g(-1) to 1900 ng g(-1), PCBs from non-detect to 47 ng g(-1), chlordanes from 1.8 ng g(-1) to 60 ng g(-1), and DDTs from non-detect to 76 ng g(-1). Consistently higher PAH concentrations found in PS foam samples (300-1900 ng g(-1)) led us to examine unexposed PS foam packaging materials and PS virgin pellets. Unexposed PS foam contained higher concentrations of PAHs (240-1700 ng g(-1)) than PS virgin pellets (12-15 ng g(-1)), suggesting that PAHs may be produced during manufacturing. Temporal trends of debris were investigated at one site, Ocean Beach, where storm events and beach maintenance were found to be important variables influencing debris present at a given time.

The histologic spectrum of epithelial neoplasms induced by sorafenib.

Sorafenib is a multikinase inhibitor newly approved for the treatment of renal cell carcinoma and hepatocellular carcinoma. Multiple cutaneous adverse effects of sorafenib have been described. We present a 68-year-old patient with renal cell carcinoma who developed multiple tender hyperkeratotic papules within weeks of starting sorafenib. The degree of symptoms and size of lesions corresponded directly with his sorafenib dosing. Four biopsy specimens of representative lesions were taken. Three lesions showed keratin-filled endophytic epithelial-lined invaginations, one with a coexistent actinic keratosis. The fourth biopsy specimen revealed an invasive squamous cell carcinoma with keratoacanthoma-like features. To our knowledge, diffuse eruptions of epidermal invaginations, ectatic follicular infundibula, and follicular infundibular cysts have not been reported previously with sorafenib, although they are well known to occur with epidermal growth factor receptor inhibitor therapy. Keratoacanthoma and squamous cell carcinoma as a result of sorafenib use are only beginning to be reported in the literature. At the time of acceptance of our manuscript, sorafenib-induced keratoacanthoma was noted only once in the literature, and deeply invasive squamous cell carcinoma has been reported once in the setting of sorafenib and tipifarnib combination therapy. We review the spectrum of dermatologic side effects of sorafenib to facilitate their recognition.

Gonadectomy and androgen replacement alter cardiac performance in conscious adult male rats.

Gender disparities in cardiac function have been described. Yet the extent to which gender related differences in cardiac performance are due to the presence of sex-specific biological factors are unclear. We used a longitudinal study aimed at examining whether castration and androgen replacement affects cardiac performance in conscious adult male rats. Adult male rats were implanted with Piezoelectric transit-time gauges and radio telemetry devices to measure regional myocardial segment length and hemodynamic variables before and after castration and after androgen replacement. Androgen withdrawal accelerated average heart rates by 7% (p=0.010). Heart rate was lowered to intact values when androgens were restored to normal physiological levels (p=0.004). Mean arterial pressure was not affected by androgen deprivation and androgen replacement. However, androgen withdrawal produced a 40% decrease in the velocity of circumferential shortening and a 46% reduction in the rate of myocardial relaxation. Androgen supplementation completely restored contractile function. These results provide the first evidence that androgen withdrawal and androgen replacement produces dramatic alterations on cardiac performance in conscious animals and demonstrates the significance of androgens as a cardio-regulatory hormone in males. Sex steroids are likely contributors to gender-related differences in cardiac function.

Acute actions of testosterone on contractile function of isolated rat ventricular myocytes.

Variation between the sexes in cardiac function have been established. The extent to which sex hormones are responsible for these differences is unclear. The current study was designed to determine whether testosterone acts acutely to enhance contractility of cultured rat ventricular myocytes. Following a 24-h treatment with testosterone (1 microM), isolated rat ventricular myocytes display a 21% increase (P < 0.01) in peak shortening and an 18% decrease (P < 0.02) in time to peak shortening. In accordance with this change, testosterone treatment produced an 18% decline (P < 0.002) in the time to relengthening when compared to vehicle-treated controls. These results provide the first evidence that short-term androgen exposure acts directly to stimulate contractility of isolated rat ventricular myocytes and thus may play a role in regulating cardiac performance in males and thereby contribute to sex differences in cardiac function.

Inhibition of sarco(endo)plasmic reticulum Ca2+-ATPase differentially regulates contractile function in cardiac myocytes from normotensive and spontaneously hypertensive rats: role of Ca2+ regulatory proteins.

Hypertension leads to impaired contractile function. This study examined the impact of inhibition of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) by thapsigargin or cyclopiazonic acid (CPA) on cardiac contractile function in ventricular myocytes from Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). Mechanical properties were examined including peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR90), and maximal velocity of shortening/relengthening (+/-dL/dt). Intracellular Ca2+ transients were evaluated as fura-2 fluorescent intensity (FFI), excitation-induced change in FFI (DeltaFFI = peak-basal), and fluorescence decay rate (tau). Expression of Ca2+ regulatory proteins SERCA2a, Na+-Ca2+ exchanger (NCX), and phospholamban (PLB) were assessed by reverse transcriptase polymerase chain reaction and Western blot. SHR rats exhibited elevated blood pressure. SHR myocytes displayed decreased PS +/- dL/dt, peak FFI, and DeltaFFI; shortened TPS; prolonged tau with normal TR90; and basal FFI compared with WKY myocytes. Inhibition of SERCA with thapsigargin (5 microM) or CPA (10 microM) significantly depressed PS +/- dL/dt, baseline FFI, and DeltaFFI, and prolonged TPS, TR90, and tau in WKY myocytes. However, SHR myocytes were relatively insensitive to thapsigargin or CPA with only TPS and TR90 prolonged. Both mRNA and protein expressions of NCX and PLB were significantly enhanced, whereas SERCA2a protein abundance was reduced in SHR rats compared with the WKY group. Our data suggest that inhibition of SERCA function differentially affected cardiac contractile function in ventricular myocytes from normotensive and hypertensive rats possibly through reduced SERCA2a, elevated PLB, and NCX expression under hypertension.

Gonadectomy alters myosin heavy chain composition in isolated cardiac myocytes.

Sex differences in cardiac function have been identified. Studies suggest that the presence of testosterone in males may contribute to the observed differences in cardiac function. Our laboratory has shown previously that testosterone treatment of gonadectomized adult male rats enhances contractility of isolated rat ventricular myocytes. In this study we tested the hypothesis that gonadectomy and hormone replacement influences contractility by altering myosin heavy chain (MHC) composition. To test this hypothesis we analyzed myosin isoform expression in ventricular myocytes isolated from castrated rats displaying a decrease in myocyte contractile velocity and compared them to castrates treated with testosterone that displayed normal myocyte shortening velocity. Sixteen weeks after castration isolated rat ventricular myocytes displayed a 90% (p < 0.001) decline in MHC-alpha mRNA levels and over a twofold (p < 0.01) increase in MHC-beta transcripts when compared to sham-operated controls. Consistent with these changes we also observed a substantial decline in the ratio of MHC-alpha to MHC-beta protein expression. A reversal in myosin heavy chain composition was achieved following testosterone replacement. These studies provide the first direct evidence that testosterone replacement in gonadectomized animals enhances contractility via transcriptional and translational control of myosin heavy chain composition in isolated rat ventricular myocytes. The influence of testosterone on MHC composition in males may underlie some of the observed sex differences in cardiac function.

Gonadectomy of adult male rats reduces contractility of isolated cardiac myocytes.

Sex-related differences in cardiac function have been well documented. The extent to which sex hormones are responsible for these differences is unclear. The current study was designed to determine whether castration and androgen replacement resulted in changes in functional expression of genes encoding the L-type calcium channel and Na/Ca exchanger in isolated rat ventricular myocytes. Sixteen weeks of castration produced a 50% decline in dihydropyridine receptor expression levels and a 16% (P < 0.05) increase in time to peak shortening. Furthermore, cardiac myocytes isolated from castrated animals also displayed an 18% (P < 0.001) increase in time to relengthening and an 80% decrease in Na/Ca exchanger gene expression when compared with intact controls. Testosterone treatment of castrated animals completely reversed these effects. These results provide the first evidence that androgens regulate functional expression of the L-type calcium channel and the Na/Ca exchanger in isolated rat ventricular myocytes and thus may play a role in modulating cardiac performance in males and thereby contribute to the observed gender differences in cardiac function.

Norepinephrine regulates the in vivo expression of the L-type calcium channel.

The alpha1c subunit (DHP receptor) of the L-type Ca2+ channel is important for calcium homeostasis in cardiac muscle. The DHPr provides the primary mechanism for calcium influx during contraction. Published results demonstrate three in vitro signaling pathways that are important in the regulation of DHPr gene expression in neonatal cardiac myocytes, the protein kinase A (PKA), protein kinase C (PKC) pathways, and intracellular calcium. To determine whether these pathways are important in vivo, we treated adult rats with infusions of isoproterenol, or norepinephrine at 200 microg/kg/h and assessed DHPr mRNA and protein levels. Following a 3-day infusion isoproterenol (ISO) and norepinephrine (NE) produced a small but insignificant reduction in DHPr mRNA levels. When the infusions were continued for 7 days isoproterenol increased DHPr mRNA accumulation to control levels while NE stimulated a 35% increase in DHPr mRNA levels and a 35% increase in protein abundance when compared to controls (p < 0.05). Furthermore, contractility and Ca2+ transient measurements of isolated cardiac myocytes from NE infused animals also display shortened duration of contraction/relaxation and increased intracellular free Ca2+ (DFFI) in response to electrical stimulation (p < 0.01). We conclude norepinephrine treatment alters DHPr mRNA and protein levels, and augments excitation-contraction coupling, and thus may be important for modulating cardiac calcium homeostasis in vivo.

Effect of insulin on iodide uptake in mouse mammary gland explants.

Studies were carried out primarily to assess the role of insulin in regulating iodide uptake in the mammary gland. Using cultured mammary gland explants from virgin and pregnant mice (12-14 days into gestation), insulin (1 microg/ml) was shown to stimulate iodide uptake after a 2-day exposure period. The effect of insulin was manifested by itself, as well as in the presence of cortisol and prolactin. Optimal iodide uptake was observed when tissues were treated with all three lactogenic hormones (insulin, cortisol, and prolactin). In a time-course experiment, the effect of insulin alone was initially observed after a 10-hr treatment; the effect was maintained for 30 hr. In dose-response studies, 1 ng/ml insulin elicited a significant effect after 24 hr in culture; a maximal effect was achieved with 50-100 ng/ml insulin. The optimal cortisol concentration for a maximum stimulation of iodide uptake was 10(-7)M. In a quantitative Western blot analysis employing an antibody to the sodium-iodide symporter, insulin stimulated an upregulation of the transporter protein after a 4-, 8-, or 20-hr treatment with insulin. Perchlorate and thiocyanate abolished the insulin effect on iodide uptake, further suggesting that the insulin response occurs via a stimulation of the sodium-iodide symporter. Clearly, insulin is an important and essential hormone in the lactogenic hormone complex for regulating iodide uptake in the mammary gland, but maximal expression of iodide uptake is only expressed when all three lactogenic hormones are present.

Castration reduces mRNA levels for calcium regulatory proteins in rat heart.

Sex-related differences in the cardiac phenotype have been well established. This study was designed to determine whether androgens regulate myocardial gene expression and play a role in the sex-related differences in the myocardial phenotype. Gonadectomized male rats were treated with testosterone, and myocardial gene expression was examined in whole heart using quantitative real-time PCR. Gonadectomy produced a substantial decrease in mRNA levels for the androgen receptor, Na(+)/Ca(2+) exchanger, L- type calcium channel, and beta(1)-adrenergic receptor (beta(1)AR). Supplementation of testosterone in castrates produced a fivefold increase in androgen receptor mRNA levels. Testosterone treatment of castrates produced almost a sixfold increase in Na(+)/Ca(2+) exchanger mRNA, a tenfold increase in Ltype calcium channel mRNA accumulation, and a fourfold increase in beta(1)AR mRNA levels. Increased calcium channel expression, beta(1)AR expression, and Na(+)/Ca(2+) exchanger expression together may alter cytosolic calcium. These results provide the first evidence that testosterone regulates expression of myocardial calcium regulating genes and thus may play a role in modulating the cardiac phenotype in males.

Random walk in an eddy and tube formation from fine particles.

Tubular shape formation of an ensemble of ultrafine particles, captured by microscopic eddies in a fluid or gaseous medium, is investigated. In the circulation flow of the eddy, the small particles are driven by the deterministic hydrodynamical forces and the random forces of Brownian motion. The conditions for dynamically/statistically stable tube formation and the resulting tube parameters are obtained by analytic calculations and computer simulations, respectively. The model yields striking similarities to the characteristics of nanotube formation observed in turbulent media such as the carbon arc, and throws some light on tubular formation observed in fluid media. (c) 2001 American Institute of Physics.

Multicenter evaluation of the performance and clinical utility in longitudinal monitoring of the Bayer Immuno 1 complexed PSA assay.

We conducted a multicenter evaluation of the analytical and clinical performance of the automated Bayer Immuno 1 complexed PSA (cPSA) assay, and compared assay performance to the Bayer Immuno 1 PSA assay. We sought to determine whether measurements of cPSA could be of clinical utility in the management of patients with prostate cancer. Results of the 10-day imprecision across three evaluation sites produced total CV < 2.50% and an analytical sensitivity of 0.02 microgram/L. There was an increased trend in clinical sensitivity for prostate cancer with increasing stage of disease (71-86%). Clinical specificity for patients with benign urogenital disease was 74.8%, and for other nonprostate diseases ranged from 91.1-100%. Retrospective serial monitoring of 155 patients with prostate cancer demonstrated concordance of cPSA measurements to clinical status for 97% of the patients analyzed. Results from the clinical studies using the Bayer Immuno 1 cPSA assay were comparable to results obtained with the Bayer Immuno 1 PSA assay. The Bayer Immuno 1 cPSA assay demonstrates analytical performance and clinical effectiveness in the management of prostate cancer patients during the course of disease and therapy.

Multiple single-unit recordings in the striatum of freely moving animals: effects of apomorphine and D-amphetamine in normal and unilateral 6-hydroxydopamine-lesioned rats.

Ensembles of striatal neurons were recorded in freely moving normal and unilateral 6-hydroxydopamine (6-OHDA)-lesioned rats using chronically implanted electrode arrays. Animals received bilateral striatal implants of two 16-microwire arrays 1 week before recordings. Identified striatal neurons were categorized as medium spiny-like and large aspiny-like based on a combination of their activity autocorrelations and firing rates. Baseline firing rates of medium spiny-like neurons in the 6-OHDA-lesioned striata were significantly faster than were firing rates of the same neurons in the intact hemispheres of 6-OHDA-lesioned rats or normal animals. However, firing rates of large aspiny-like neurons were faster in both hemispheres of the 6-OHDA-lesioned rats as compared to normal animals. Interestingly, firing rates of neurons in all groups decreased by fivefold or greater under urethane anesthesia, although the relative firing rates between hemispheres were unchanged. d-Amphetamine (5.0 mg/kg, s.c.) increased the firing rates of both types of striatal neurons by twofold or greater in normal rats and in the intact hemispheres of 6-OHDA-lesioned animals. By contrast, this treatment did not alter neuron firing in the 6-OHDA-lesioned striata. Apomorphine (0.05 mg/kg, s.c.) did not affect neuronal firing rates either in normal rat striatum or in the unlesioned hemispheres of 6-OHDA-lesioned animals. However, it did significantly increase the firing rate of the medium spiny-like neurons in 6-OHDA-lesioned striata. These results demonstrate that the dopaminergic innervation of the striatum differentially influences two electrophysiologically distinct sets of striatal neurons in freely moving rats.

Multicenter evaluation of the Bayer Immuno I CA 15-3 assay.

We conducted a multicenter evaluation of the analytical and clinical features of the automated Bayer Immuno 1 CA 15-3 assay and compared assay performance to two manual tests. Results of the 10-day imprecision study of the Bayer Immuno 1 assay pooled across four evaluation sites and three lots of reagent produced total CV < or = 4%. Lot-to-lot reproducibility for 26 different lots of reagents and calibrators manufactured over a 2-year period was demonstrated (CV, 1.1%). Results for the Bayer Immuno 1 assay correlated well with the Biomira TRUQUANT BR 27.29 and Centocor CA 15-3 RIAs (r > or = 0.94). The upper limit of the reference interval for the Bayer Immuno 1 assay was 35.9 kilounits/L (35.9 units/mL); values were similar for all methods. Longitudinal monitoring of healthy women yielded assay values with an average CV of 11% and 21% for the Bayer Immuno 1 and Biomira assays, respectively. The Bayer Immuno 1 assay demonstrated the analytical features, intermethod correlation, and long-term performance characteristics that are essential for longitudinal monitoring of breast cancer patients.

Bayer Immuno 1 PSA Assay: an automated, ultrasensitive method to quantitate total PSA in serum.

The Bayer Immuno 1 PSA Assay measures total PSA in human serum and demonstrates excellent performance with an interassay CV < or = 3.4% and a biological detection limit of 0.03 microgram/L. No significant interference from common hormonal and chemotherapeutic drugs, kallikrein, prostatic acid phosphatase, and trypsin, or elevated levels of total bilirubin, hemoglobin, triglycerides, and IgG was observed. The 95th percentile values for healthy individuals increased with age from 3.0 micrograms/L for males 50-59 years and 3.3 micrograms/L for males 60-69 years, to 4.6 micrograms/L for males > or = 70 years. Clinical studies with retrospective samples demonstrated correspondence between serial measurements of PSA and clinical outcome for 98% of 159 prostate cancer patients. Clinical sensitivity for patients with clinical evidence of disease, untreated at the time of specimen draw, increased with increasing stage from 77.5-100%. Specificity of 60-70% for BPH and other benign urogenital diseases was consistent with previous findings. Bayer Immuno 1 PSA Assay values for 2131 specimens from healthy subjects and patients with prostate cancer, BPH, and other malignant and nonmalignant diseases correlated well with the Abbott IMx PSA Assay over the range 0.0-6,238 micrograms/L (Y = 1.10 x + 0.02). The Bayer Immuno 1 PSA Assay provides automated ultrasensitive, precise, and equimolar measurement of total PSA in human serum.

The SH3 domain of p56lck binds to proline-rich sequences in the cytoplasmic domain of CD2.

CD2, a cell surface glycoprotein expressed on T cells and natural killer cells, can couple to signaling pathways that result in T cell proliferation. An Src-like protein tyrosine kinase, p56lck, coprecipitates with CD2, and perturbation of CD2 by monoclonal antibodies results in an increase in the activity of p56lck, suggesting that an interaction with p56lck contributes to CD2-mediated signaling. Herein, we investigate the mechanism by which CD2 associates with p56lck. We demonstrate that CD2 and p56lck associate when coexpressed in nonlymphoid cells, that this association requires the cytoplasmic domain of CD2, and that the SH3 domain of p56lck mediates its interactions with CD2. Using truncation mutants of CD2, we identify two regions in the cytoplasmic domain of CD2 involved in binding p56lck. Each region contains a proline-rich sequence that, in the form of a synthetic peptide, directly binds p56lck. Thus, proline-rich sequences in the cytoplasmic domain of CD2 allow this transmembrane receptor to bind to the SH3 domain of p56lck.

Multipopulation survey designs: five types with seven shared aspects.

"Five types of multipopulation surveys are defined and described: periodic surveys; multidomain designs; multinational survey designs; cumulated and combined samples; and controlled observations or quasi- experimental designs. I emphasize the deliberate designs of these surveys, not the mere post hoc or ad hoc utilization of survey results. Most importantly, I emphasize a sharp distinction between seven listed aspects. Similarity and standardization of the survey aspects (definitions, methods, measurements) are essential to avoid biases in comparisons, though admittedly difficult. In contrast, flexibility in sampling designs and sizes, to reduce variances, is permissible, also desirable to reduce variances and costs." (SUMMARY IN FRE)

Statistical medicine.

In the Perspective by Donald G. York (8 July, p. 191), figure 1 was printed incorrectly. The correct figure and caption appear below. [See figure in the PDF file]

The effect of cycloplegia on measurement of the ocular components.

PURPOSE: The purpose of this study was to examine the effect of cycloplegic agent on the measurement of refractive error and the ocular components. METHODS: We compared two commonly used topical cycloplegic agents, 1% tropicamide and 1% cyclopentolate, for their effect on the measurement of refractive error (by Canon R-1 autorefraction), accommodative response (by Canon R-1 autorefraction and by the conventional, subjective "pushup" method), crystalline lens power (by video phakometry and by calculation), and axial ocular dimensions (by A-scan ultrasonography) in 20 emmetropic to moderately hyperopic children. RESULTS: Comparison of refractive error at each drug's reported time of maximum cycloplegia (30 minutes for tropicamide and 60 minutes for cyclopentolate) showed that distance autorefraction in the vertical meridian differed by +0.20 +/- 0.30 diopters (D) (P = 0.008). The average difference was +0.07 +/- 0.10 mm for anterior chamber depth (P = 0.004), -0.03 +/- 0.05 mm for crystalline lens thickness (P = 0.025), -0.65 +/- 0.69 D for phakometrically measured crystalline lens power (P < 0.001), +0.03 +/- 1.55 D for calculated crystalline lens power (P = 0.94), and -0.09 +/- 0.19 mm for vitreous chamber depth (P = 0.062, all paired t tests; positive signs denote greater values with cyclopentolate). Residual accommodation was 0.47 and 0.67 D greater with tropicamide when measured by autorefraction and the pushup method (P = 0.013 and 0.08 respectively, paired t test). All significant differences were consistently in the direction of poorer cycloplegia with tropicamide. CONCLUSIONS: Although tropicamide, as expected, showed poorer cycloplegia compared to cyclopentolate, the degree of difference appeared to be small, with minimal effect on the measurement of distance refractive error and the ocular optical components.