Prospective monitoring of adverse events following vaccination with Modified vaccinia Ankara - Bavarian Nordic (MVA-BN) administered to a Canadian population at risk of Mpox: A Canadian Immunization Research Network study.

MVA-BN is an orthopoxvirus vaccine that provides protection against both smallpox and mpox. In June 2022, Canada launched a publicly-funded vaccination campaign to offer MVA-BN to at-risk populations including men who have sex with men (MSM) and sex workers. The safety of MVA-BN has not been assessed in this context. To address this, the Canadian National Vaccine Safety Network (CANVAS) conducted prospective safety surveillance during public health vaccination campaigns in Toronto, Ontario and in Vancouver, British Columbia. Vaccinated participants received a survey 7 and 30 days after each MVA-BN dose to elicit adverse health events. Unvaccinated individuals from a concurrent vaccine safety project evaluating COVID-19 vaccine safety were used as controls. Vaccinated and unvaccinated participants that reported a medically attended visit on their 7-day survey were interviewed. Vaccinated participants and unvaccinated controls were matched 1:1 based on age group, gender, sex and provincial study site. Overall, 1,173 vaccinated participants completed a 7-day survey, of whom 75 % (n = 878) also completed a 30-day survey. Mild to moderate injection site pain was reported by 60 % of vaccinated participants. Among vaccinated participants 8.4 % were HIV positive and when compared to HIV negative vaccinated individuals, local injection sites were less frequent in those with HIV (48 % vs 61 %, p = 0.021), but health events preventing work/school or requiring medical assessment were more frequent (7.1 % vs 3.1 %, p = 0.040). Health events interfering with work/school, or requiring medical assessment were less common in the vaccinated group than controls (3.3 % vs. 7.1 %, p < 0.010). No participants were hospitalized within 7 or 30 days of vaccination. No cases of severe neurological disease, skin disease, or myocarditis were identified. Our results demonstrate that the MVA-BN vaccine appears safe when used for mpox prevention, with a low frequency of severe adverse events and no hospitalizations observed.

Metabolic Engineering of a Pentose Metabolism Pathway in Ethanologenic Zymomonas mobilis.

The ethanol-producing bacterium Zymomonas mobilis was metabolically engineered to broaden its range of fermentable substrates to include the pentose sugar xylose. Two operons encoding xylose assimilation and pentose phosphate pathway enzymes were constructed and transformed into Z. mobilis in order to generate a strain that grew on xylose and efficiently fermented it to ethanol. Thus, anaerobic fermentation of a pentose sugar to ethanol was achieved through a combination of the pentose phosphate and Entner-Doudoroff pathways. Furthermore, this strain efficiently fermented both glucose and xylose, which is essential for economical conversion of lignocellulosic biomass to ethanol.

Interferon inducers in vitro: difference in sensitivity to inhbitiros of RNA and protein synthesis.

Interferon can be induced by diverse agents in a variety of mammalian cell cultures through apparently two mechanisms. One results in an early (2 to 10 hours) appearance of interferon and is relatively resistant to inhibition by actinomycin, puromycin, or fluorophenylalanine. A second mechanism results in a late (18 to 24 hours) appearance of interferon and is more sensitive to inhibition by these inhibitors. The molecular basis for each mechanism is unclear. Since each interferon inducer may have multiple effects on the cell, the differences observed may not necessarily reflect a fundamental difference in the mechanism of interferon stimulation.

Modeling the intraurban variability of ambient traffic pollution in Toronto, Canada.

The objective of this paper is to model determinants of intraurban variation in ambient concentrations of nitrogen dioxide (NO2) in Toronto, Canada, with a land use regression (LUR) model. Although researchers have conducted similar studies in Europe, this work represents the first attempt in a North American setting to characterize variation in traffic pollution through the LUR method. NO2 samples were collected over 2 wk using duplicate two-sided Ogawa passive diffusion samplers at 95 locations across Toronto. Independent variables employed in subsequent regression models as predictors of NO2 were derived by the Arc 8 geographic information system (GIS). Some 85 indicators of land use, traffic, population density, and physical geography were tested. The final regression model yielded a coefficient of determination (R2) of .69. For the traffic variables, density of 24-h traffic counts and road measures display positive associations. For the land use variables, industrial land use and counts of dwellings within 2000 m of the monitoring location were positively associated with NO2. Locations up to 1500 m downwind of major expressways had elevated NO2 levels. The results suggest that a good predictive surface can be derived for North American cities with the LUR method. The predictive maps from the LUR appear to capture small-area variation in NO2 concentrations. These small-area variations in traffic pollution are probably important to the exposure experience of the population and may detect health effects that would have gone unnoticed with other exposure estimates.

Enzyme replacement via liposomes. Variations in lipid compositions determine liposomal integrity in biological fluids.

Liposomes survive exposure to biological fluids poorly, extruding trapped enzymes, drugs, or solutes upon interaction with serum or plasma constituents. We have quantified the disruptive effects of human serum on liposomes and have studied whether various modifications in their phospholipid composition might produce liposomes with an increased carrier potential for application in vivo. Multilamellar liposomes (phosphatidycholine 70:dicetyl phosphate 20:cholesterol 10) were prepared with 3H-labeled phosphatidylcholine as the lipid phase marker and [14C]inulin and horseradish peroxidase as aqueous phase markers. Gel exclusion chromatography showed that 32 +/- 3% of [14C]inulin and 27 +/- 7% of horseradish peroxidase were lost after 1 h incubation with 10% (v/v) human serum. Loss of aqueous solutes was reduced to 20 +/- 5%/h and 17 +/- 2%/h, respectively, after treatment with decomplemented serum (56 degrees C, 30 min). Loss induced by serum was concentration and time dependent: to 57 +/- 2% at 1 h and 67 +/- 14% at 24 h, with 50% serum; plasma was slightly less perturbing whereas human serum albumin was not at all disruptive. By incorporating sphingomyelin (35 mol%) into multilamellar liposomes, the leakage of [14c]-inulin in the presence of 10% serum was reduced to 12 +/- 4%/h; increasing the molar percentage of cholesterol to 35% also stabilized the lipid bilayers, reducing leakage to 20 +/- 7%/h. Both small and large unilamellar vesicles could not be stablilized against serum-mediated leakage by the incorporation of sphingomyelin. The data suggest that cholesterol and sphingomyelin enhance liposomal integrity in the presence of serum or plasma and promise to yield enhanced survival of drug-laden lipid vesicles in biological fluids in vivo.

Liposome uptake by human leukocytes. Enhancement of entry mediated by human serum and aggregated immunoglobulins.

The entry of immunoglobulin-coated liposomes into human leukocytes bearing Fc receptors was evaluated using two methods: (i) the cellular association of liposomal markers (3H-labelled phosphatidylcholine, lipid phase; [14C]inulin, aqueous phase), and (ii) the ultrastructural cytochemistry of cells following incubation of cells with liposomes containing a cytochemical marker (horseradish peroxidase) in the aqueous spaces. The entry of liposomes into a cell population composed predominantly of neutrophils was linear for 10--15 min and was mediated by an active process that appeared to be both energy- and surface-dependent. This uptake could be largely inhibited by incubation at 0 degrees C, and by exposure to glutaraldehyde, iodoacetamide, N-ethylmaleimide, and an excess of aggregated immunoglobulins. Entry into cells of multilamellar liposomes was saturable, displaying affinity constants of 1.1 and 1.7 mM. Ultrastructural analysis of the heterogeneous leukocyte population showed that monocytes took up liposomes more actively than neutrophils and lymphocytes. Moreover, liposomes were almost always found within the leukocytes, rather than adherent to the outer plasma membrane. The relative avidity of monocytes was confirmed by comparing the uptake of radiolabelled liposomes by a 'pure' neutrophil population, a 'mixed' neutrophil population, and a 'mononuclear cell' population. Precoating liposomes with high molecular weight aggregates of human immunoglobulin G resulted in enhanced serum-independent uptake. The fraction of aggregated immunoglobulin G which was most effective in provoking uptake of coated liposomes also stimulated the greatest amount of lysosomal enzyme secretion. These data suggest that the interaction (precoating) of liposomes with either high molecular weight aggregates of immunoglobulin G or with serum enhances their subsequent uptake by human leukocytes.

Comparison of large unilamellar vesicles prepared by a petroleum ether vaporization method with multilamellar vesicles: ESR, diffusion and entrapment analyses.

Large unilamellar vesicles, prepared by a petroleum ether vaporization method, were compared to multilamellar vesicles with respect to a number of physical and functional properties. Rotational correlation time approximations, derived from ESR spectra of both hydrophilic (3-doxyl cholestane) and hydrophobic (3-doxyl androstanol) steroid spin probes, indicated similar molecular packing of lipids in bilayers of multilamellar and large unilamellar liposomes. Light scattering measurements demonstrated a reduction in apparent absorbance of large unilamellar vesicles, suggesting loss of multilamellar structure which was confirmed by electron microscopy. Furthermore, large unilamellar vesicles exhibited enhanced passive diffusion rates of small solutes, releasing a greater percentage of their contents within 90 min than multilamellar vesicles, and reflecting the less restricted diffusion of a unilamellar system. The volume trapping capacity of large unilamellar vesicles far exceeded that of multilamellar liposomes, except in the presence of a trapped protein, soy bean trypsin inhibitor, which reduced the volume of the aqueous compartments of large unilamellar vesicles. Finally, measurement of vesicle diameters from electron micrographs of large unilamellar vesicles showed a vesicle size distribution predominantly in the range of 0.1--0.4 micron with a mean diameter of 0.21 micron.

Interaction of liposomes with human leukocytes in whole blood.

The uptake of multilamellar liposomes into human leukocytes in whole blood in vitro was evaluated on the basis of the cellular association of liposomal markers (3H-labelled cholesterol, lipid phase; [14C]inulin, aqueous phase). The entry of liposomes into human blood leukocytes was linear for 60 min and was mediated by a saturable mechanism displaying affinity constants of 0.28 +/- 0.17 and 0.16 +/- 0.05 mM liposomal lipid (means +/- S.E.) for liposomal lipid and aqueous phase markers, respectively. Amicon filtration analysis of incubation mixtures containing blood and liposomes (phosphatidylcholine:dicetyl phosphate:cholesterol, 70:20:10) showed that 34% of [14C]inulin was lost (neither liposome-associated nor cell-associated) after 60 min. By preincorporating sphingomyelin (35 mol%) into multilamellar liposomes, the leakage of the model aqueous phase marker inulin was reduced to 8% after 60 min, thus enhancing the drug carrier potential of liposomes in blood. As a consequence of their interaction with liposomes, the polymorphonuclear leukocytes in whole blood decreased in apparent buoyant density, while maintaining their viability. These results indicate that blood leukocytes in their natural milieu of whole blood are capable of interacting with, and taking up multilamellar liposomes.

Lifesaving explains mortality decline with time.

Mortality rates of human populations in developed countries are declining with time. We show that this effect can be explained via a 'lifesaving' methodology. Our approach is based on considering a non-homogeneous Poisson process of potentially harmful events. Each of these events can be 'cured' with a given probability or can result in a termination of the Poisson process (death) with a complementary probability. A lifesaving ratio, defining the corresponding relative increase in life expectancy for homogeneous and heterogeneous populations is analyzed. Some generalizations are discussed. Several simple examples are considered.

Difficulties in estimating glomerular filtration rate in the elderly.

Estimates of glomerular filtration rate are generally obtained by measuring or estimating endogenous creatinine clearance. However, it may sometimes be difficult to obtain the necessary urine collections. Most of 19 healthy, reliable elderly outpatients were found unable to provide satisfactory 24-hour urine collections. To judge whether formulas estimating creatinine clearance from serum creatinine levels are reliable, we also compared 24-hour creatinine clearances measured in 50 inpatients with values calculated by the Cockroft-Gault equation. Only a moderate correlation was found, which may be unacceptable in the clinical situations for which the equation is used, such as drug dosing. For reasons including uncertainties in the validity of predictive formulas and unreliability of urine collections, we conclude that no acceptable method now exists for bedside estimation of glomerular filtration rate and that drug levels should be measured whenever possible in elderly patients and in those with renal insufficiency.

Comparative studies of the aromatization of testosterone and epitestosterone by human placental aromatase.

The aromatization of epitestosterone (17 alpha-hydroxy-4-androsten-3-one) and testosterone by lyophilized human placental microsomes was studied. Upon incubation of epitestosterone, 12% was converted to 17 alpha-estradiol, 15% to 19-keto-epitestosterone (17 alpha-hydroxy-4-oxo-4-androsten-19-al), 10% to 19-hydroxyepitestosterone (17 alpha, 19-dihydroxy-4-androsten-3-one), and about 10% to several unidentified products. A similar incubation with testosterone resulted in 60% conversion to 17 beta-estradiol; 30% was unchanged. At increasing substrate concentrations (0.1-50 microM), the aromatization rate of epitestosterone increased gradually and did not reach a plateau, whereas aromatization rate of testosterone plateaued at about 3 microM. The presence of either testosterone or 17 beta-estradiol in concentrations 0.1-10 times the concentration of epitestosterone inhibited the aromatization of epitestosterone by about 70%, while the aromatization of testosterone was not inhibited by either epitestosterone or 17 alpha-estradiol. Lyophilization of fresh microsomes or storage of the lyophilized microsomes at -20 C greatly reduced the aromatizing activity upon epitestosterone but not upon testosterone. These results suggest that the aromatizing system for epitestosterone is different from that for testosterone.

11 beta-Hydroxylase with affinity to C-21-Deoxysteroids from ovaries of patients with polycystic ovary syndrome.

When ovarian mitochondria from a patient with polycystic ovary syndrome (POS) were incubated with [7-3H]17alpha-hydroxypregnenolone and [4-14C]17alpha-hydroxyprogesterone, 11beta-hydroxylated metabolites were obtained. When the same mitochondrial preparation was incubated with [7-3H]17alpha-hydroxypregnenolone and [4-14C]11-deoxycortisol no 11beta-hydroxylated derivatives of 11-deoxycortisol were found. These results are compatible with and support the conclusion that the ovary of POS patients who excrete pregnanetriolone contains an 11beta-hydroxylase capable of hydroxylating C-21-deoxysteroids but not C-21-hydroxysteroids.

A cAMP-triggered release of a hormone-like peptide.

Preparations of the catalytic subunit of cAMP-dependent protein kinase from rabbit skeletal muscle, which appear to be homogeneous by SDS-polyacrylamide gel electrophoresis, were often found to contain a hormone-like factor (HLF) which causes an immediate rise, then a decline of intracellular cAMP in a B-lymphoma cell line. Active HLF is released when the fractions that contain it in an inactive form are incubated with cAMP prior to chromatography, or passed through an immobilized cAMP column. HLF seems to be a peptide: it loses its cell-stimulating capability after proteolysis and has an apparent molecular mass of 2.2-2.5 kDa.

Cost-effective imaging approach to the nonbilious vomiting infant.

OBJECTIVE: To develop a cost- and time-effective algorithm for differentiating hypertrophic pyloric stenosis (HPS) from other medical causes of emesis in infants referred from community-based pediatricians and family practitioners to the imaging department of a tertiary children's care facility. METHODS: Eighty-nine vomiting infants (22 females, 67 males) between the ages of 11 and 120 days (mean, 43.5 days) had received nothing by mouth for at least 1 hour before the study. Each child was assessed for duration of vomiting, status of body weight, time and volume of last ingestion, and time of last emesis. A #8 French (Sherwood Medical, St Louis, MO) nasogastric feeding tube was placed in the child's stomach. The contents were aspirated and measured to determine likelihood of HPS. An aspirated volume >/=5 mL implicated gastric outlet obstruction, and ultrasonography (US) was performed. If this study was positive for HPS, the patient was referred for surgery. If US was negative, an upper gastrointestinal series (UGI) was performed. An aspirated stomach contents volume <5 mL suggested a medical cause for the emesis, and UGI was performed. Pediatric surgeons with no knowledge of the volume results palpated the abdomens of 73 of 89 infants (82%). RESULTS: Twenty-three of 89 patients (25%) had HPS. The aspirate criteria for HPS had a sensitivity of 91%, a specificity of 88%, and an accuracy of 89%. Of the false-positive studies (total = 8), six were related to recent significant ingestion (within 2 hours of the study), and two were attributable to antral dysmotility. The surgeons palpated the mass in 10 of 19 patients (53%). Sensitivity and specificity were 53% and 93%, respectively. Only 6 of 89 infants (7%) required both US and UGI to determine the etiology of the nonbilious vomiting. By performing the UGI in 66 patients, it was also found that 14% had slow gastric emptying and 79% had gastroesophageal reflux. Eighty-one percent of the gastroesophageal reflux was significant. CONCLUSION: The volumetric method of determining the proper imaging study is cost- and time-effective in the evaluation of the nonbilious vomiting infant for pyloric stenosis. If US was performed initially in all patients referred for imaging, two studies would have been performed in 68 of 89 patients (76%) to define the etiology of the emesis. Because we used the volumetric method, 62 fewer imaging studies were performed, representing a savings of $4464 and 30 hours of physician time. If children are given nothing by mouth for 3 to 4 hours before gastric aspiration, the specificity of the volumetric method improves to 94%, and the accuracy improves to 96%.

A periodicity in the response of SJL/J thymocytes to isoproterenol. Simulation by cell lines.

Thymocytes from SJL/J mice exhibit a periodicity in their response to hormonal stimulation with isoproterenol. This periodicity (5-9 days) is expressed in large changes in the intensity of the response (peak levels of intracellular cAMP which vary approximately 6-fold), and in the response pattern, i.e., in the occurrence or non-occurrence of an immediate hormone-induced desensitization. In contrast, C57BL/6 thymocytes have a homogeneous response pattern (in all cases there was an immediate desensitization). Their response does change with the same periodicity, but these changes are restricted to the intensity of response (the peak of cAMP levels varies only approximately 2-fold). Using T, B and pre-B shown that these periodic changes may be due to fluctuations in T cell subpopulations in the thymus and possibly also the infiltration of B cells into it. These observations provide a useful system for studying at the cellular and molecular level the known correlation between endocrine disorders and neoplasia in mice. On the other hand, the set of cell lines with such different response patterns to a single hormone may be used for the identification and isolation of additional cellular constituents involved in the cellular response to hormones and its immediate desensitization.

Inhibition of enteropathogenic bacteria by human milk whey in vitro.

Diarrhea caused by enteropathogenic bacteria is a leading cause of childhood mortality world-wide, particularly in less developed regions. Breast-feeding has been advocated to protect infants and children from infectious illnesses. We examined the antibacterial activity of human whey in vitro against multiple strains of the following species of enteropathogenic bacteria, Campylobacter jejuni, Escherichia coli, Salmonella typhimurium, Shigella flexneri, Shigella sonnei and Vibrio cholerae, all isolated from humans with diarrheal illness. In vitro human whey inhibited the growth of most but not all of the enteropathogenic strains tested. The test strains multiplied in both bacterial growth medium and commercial infant-feeding formula controls, with the exception of the Campylobacter strains, which were markedly inhibited in formula. These results confirm and extend previous observations that human whey contains components capable of inhibiting enteropathogenic bacteria and which may be associated with the protective effects of breast feeding. The precise mechanisms of these inhibitory effects merit further study.

The relationship between parental occupation and bone cancer risk in offspring.

BACKGROUND: Bone cancers in children are serious and highly fatal conditions, yet relatively little is known about their causes or methods of prevention. METHODS: The relationship between parental occupation and bone cancer in offspring was explored in a case-control study. Cases were identified from the Ontario Cancer Registry; population-based controls were matched on sex and age. Data were collected from their parents through the use of a mailed self-administered questionnaire. RESULTS: The odds ratio estimates (OR) for bone cancer were elevated for fathers in the social sciences (OR = 2.5, 95% confidence interval [CI]: 0.7-8.4). Risk of Ewing's sarcoma was significantly high among children with fathers in social sciences (OR = 6.2, 95% CI: 1.6-24.5) and mothers in teaching (OR = 3.1, 95% CI: 1.1-8.7) or farming (OR = 7.8, 95% CI: 1.9-31.7). Osteosarcoma risk was increased for fathers in farming (OR = 2.1, 95% CI: 0.8-5.7), and mothers in managerial and administrative work (OR = 2.3, 95% CI: 0.6-8.1), and product fabricating, assembling, and repairing (OR = 2.0, 95% CI: 0.6-7.2). CONCLUSIONS: Certain methodological problems plague studies of bone cancer in children (e.g. small studies, low statistical power, analysis of multiple occupational categories, difficulty in identifying specific carcinogenic agents). These associations require further investigation, especially as elevated risks have been reported previously for agricultural occupations.

Pneumococcal bacteremia in adults: age-dependent differences in presentation and in outcome.

In order to evaluate the effect of age on the presentation of and response to acute bacterial infection, the hospital charts of 187 adult patients with community-acquired pneumococcal bacteremia admitted to Bellevue Hospital over a nine-and-a-half year period were reviewed. Compared with younger patients, older patients (aged 65 or older) more frequently had (1) a lower fever in response to the infection, (2) an unclear history of illness, (3) a delay in diagnosis and/or therapy, and (4) a higher risk of dying. On admission, their leukocyte counts and heart rates were similar to those in a group of younger patients, which was composed largely of alcoholic patients and those addicted to intravenous drugs. Response to therapy was also similar in surviving older patients. Lower temperature and an unclear history were features most commonly associated with both delayed diagnosis and higher mortality. When patients with a history of alcohol abuse and those dying shortly after admission (i.e., presenting in a moribund state) were eliminated from the analysis, many of these age-related differences in presentation and outcome became even more evident.

Ultrasound evaluation of scrotum in pediatrics.

Scrotal abnormalities are difficult to assess using clinical criteria alone. Ultrasound provides an accurate means of demonstrating the scrotal contents so that appropriate therapy may be instituted. In a retrospective study, 119 ultrasound examinations of 96 patients (aged 4 days to 23 years) have been compared with the clinical diagnosis, surgical/pathologic findings, and other imaging modalities. The gamut of disease identified included congenital anomalies, neoplasm, trauma, torsion, varicocele, hydrocele, epididymo-orchitis, epididymal cyst/spermatocele, and post-radiation fibrosis. The ultrasound findings correlated well in 93/96 patients. In inconclusive cases, sequential imaging helps differentiate traumatic and inflammatory lesions from neoplastic processes.