Gel-entrapment of perfluorocarbons: a fluorine-19 NMR spectroscopic method for monitoring oxygen concentration in cell perfusion systems

Oxygenation is a major determinant of the physiological state of cultured cells. 19F NMR can be used to determine the oxygen concentration available to cells immobilized in a gel matrix by measuring the relaxation rate (1/T1) of perfluorocarbons (PFC) incorporated into the gel matrix. In calcium alginate gel beads without cells the relaxation rate (1/T1) of the trifluoromethyl group of perfluorotripropylamine (FTPA) varies linearly with oxygen concentration, with a slope of 1.26 +/- 0.15 x 10(-3) s-1 microM-1 and an intercept of 0.50 +/- 0.04 s-1. During perfusion with medium equilibrated with 95%/5% O2/CO2, changes in PFC T1s indicate that the average oxygen concentration was reduced from 894 +/- 102 microM in the absence of cells to 476 +/- 65 microM and 475 +/- 50 microM in the presence of 0.7 x 10(8) EMT6/Ro and RIF-1 murine tumor cells per milliliter of gel, respectively. The presence of 0.2 microliters of FTPA/ml of gel had no effect on the energy status of the cells as indicated by 31P NMR spectra. To calculate oxygen gradients within the beads from the average PFC T1 of the sample, a mathematical model was used assuming that oxygen is the limiting nutrient for cell metabolism and that the cellular oxygen consumption rate is independent of oxygen concentration. Data for EMT6/Ro cells were fit using experimentally determined perfusion parameters together with literature values for cell volume and oxygen consumption rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Association of age and body weight with periodontal disease in North American dogs

Thirteen hundred and fifty dogs were examined under anesthesia at veterinary hospitals in the USA and Canada. Periodontal health was recorded in detail. Teeth were frequently absent (particularly lower third molar, upper and lower first premolars, and incisor teeth). Calculus was most extensive on the upper fourth premolar and molar teeth. Missing teeth, mobility of remaining teeth, extent of calculus and gingival inflammation, and furcation exposure and attachment loss all were more common in small dogs compared with larger dogs, and in order dogs compared with younger dogs.     

Interpretation of chemically created periapical lesions using direct digital imaging

Perchloric acid (70%) was used to create simulated periapical lesions in tooth sockets of 15 dentate cadaver jaw specimens. Using the Trophy USA direct digital radiographic system, linear images were captured at selected time intervals after initial acid application and altered by contrast reversal, pseudocolor enhancement, and two forms of histogram equalization. The 525 total images were randomized for display on a computer monitor for evaluation by five endodontists. Images were evaluated twice by each rater, with viewings 1 to 2 wk apart. Statistical analysis determined interrater variability, intrarater reproducibility, and the relative merits of each enhancement technique. At 8, 12, 16, and 24 h after acid application, both techniques of histogram equalization yielded a statistically significant improvement over reverse contrast in perception of periapical patholais. Linear and pseudocolor-enhanced images were also significantly more diagnostic than reverse contrast at 12, 16, and 24 h. Intrarater reproducibility showed moderate agreement, but analysis showed only a fair level of interrater agreement.     

Enterocolitis associated with Clostridium perfringens infection in neonatal foals: 54 cases (1988-1997)

OBJECTIVE: To identify clinical signs, physical examination findings, results of diagnostic tests, treatments administered, and clinical outcome of neonatal foals with enterocolitis associated with Clostridium perfringens infection. DESIGN: Retrospective study. ANIMALS: 54 neonatal foals. RESULTS: Most foals had acute onset of obtunded mentation, colic, or diarrhea and developed leukopenia, neutropenia, an abnormally high number of band neutrophils, toxic WBC, and hypoproteinemia within 24 hours after admission, despite high serum IgG concentrations (> 800 mg/dl). Abdominocentesis and abdominal radiography of some foals revealed exudative peritonitis and gaseous distention of the small and large intestine, respectively. Cytologic examination of feces revealed spores or gram-positive rods in 8 of 10 foals. The most common genotypes of C perfringens isolates were type A and C, alone or in combination. Treatment did not alter mortality rate for most foals that had a positive culture for C perfringens type C. Of 54 foals, 29 (54%) that had C perfringens-associated enterocolitis died. Foals that had a culture that yielded C perfringens had higher sepsis scores, IgG concentrations, and mortality rates, compared with the overall hospital population of neonatal foals. CLINICAL IMPLICATIONS: Foals less than 7 days old that have enterocolitis associated with C perfringens infections, especially type C, have a guarded prognosis. Cytologic examination of feces to determine spore counts and detect rods may be a means for early identification of C perfringens infections. Polymerase chain reaction assays to determine genotype are important for designing preventive treatment regimens.     

Enalapril versus bendroflumethiazide in type 2 diabetes complicated by hypertension

In a double-blind, double-dummy, randomized, multi-centre study, the effects of bendroflumethiazide vs. enalapril on blood pressure, glycaemic control, lipoprotein concentrations and albuminuria were compared in non-proteinuric, hypertensive type 2 diabetic patients; they were treated for 20 weeks with either bendroflumethiazide 2.5-5.0 mg (n = 59) or enalapril 10-20 mg (n = 55). Age, fasting plasma glucose, HbA1c and BMI were similar in the groups. Systolic and diastolic blood pressure were reduced in both groups. Bendroflumethiazide was accompanied by minor but significant elevations in fasting plasma glucose and serum C-peptide. HbA1c was increased during both treatments. Lipoproteins and urinary albumin/creatinine ratio were stable. Bendroflumethiazide caused a decrease in serum potassium and an increase in serum urate. No significant correlations were observed between the decline in blood pressure and changes in the metabolic risk factors. Baseline levels of age, sex, BMI, blood pressure or urinary albumin/creatinine ratio were not related to changes in blood pressure, metabolic parameters or urinary albumin/creatinine ratio.     

Effects of a behavioural rhythm on conception probability and pregnancy outcome

This paper concerns a behaviour-driven weekday rhythm in conception rates in a large natural human population. From 1978 to 1991, North Carolina normal live single births arose from menstrual cycles which began on Monday in clear excess over other weekdays. Cycles beginning on Friday were also in excess. Cycles starting on Saturday and Sunday, or Wednesday and Thursday, each represented significantly less than one in seven of weekly totals. The source of the observed synchrony was a Sunday morning peak of coital frequency. Average cycles which began on Monday had their most fertile day on the most likely day for intercourse, translating a weekly insemination rhythm into a weekly conception rate rhythm. The secondary conception peak in Friday-onset cycles increased with age, to become the major peak for mothers aged > 30 years. We interpret this to represent a previously unreported second type of cycle with a modal follicular phase length of 10 days, the frequency of which increases with age. Several large groups of anomalous human births depart significantly from the weekday rhythm of normal conceptions. These outcomes parallel results of experimental interference with fertilization timing in estruative mammals. We believe that this implicates anomalous fertilization timing in several of the most numerous anomalies of human prenatal development.     

Evaluation of Serum Volume Losses During Long-Term Storage

Aliquots of serum collected in a large case-control study of cervical cancer were stored at −70°C for up to 4 years during implementation of the study. When 500 μL serum aliquots were thawed in preparation for carotenoid and vitamin A assays, volumes were noticeably variable and fell below 500 μL in the majority of the samples. We were concerned about evaporation/sublimation during storage of the samples because loss of water would concentrate the analytes of interest. We evaluated the use of density and sodium ion concentration measurements to confirm its occurrence. We found that serum density was an unreliable indicator of extent of volume loss since the anticipated increases in density due to evaporation were of the same magnitude as inter-individual variation in serum density. In contrast, Na⁺ concentration is tightly regulated and would rise if water had been lost from the samples. In a representative sample of serum aliquots from the case-control study, 24 of 25 vials contained less than 500 μL of serum. The mean sodium ion concentration (138.1 ± 3.6 mmol/L) was within the normal range for human serum of 136–145 mmol/L, and no correlation was observed between serum volume and Na⁺ concentration. These results strongly suggest that the observed low volumes were not due to evaporative losses. Instead, the variably low volumes of serum aliquots were probably due to pipetting errors in the initial aliquotting resulting from the use of air-displacement pipettes.     

Disappearing left atrial vegetation in an intravenous drug abuser

Isolated left atrial mural endocarditis is rare. We report a case where the diagnosis was made clinically and supported by blood cultures and transoesophageal echocardiography. Appropriate intravenous antibiotics were administered and serial transoesophageal echocardiograms helped in monitoring the decrease in size and final disappearance of the vegetation, thus avoiding the need for surgical intervention.