Automation for clinical CD4 T-cell enumeration, a desirable tool in the hands of skilled operators.

BACKGROUND: Automation in HIV clinical flow cytometry when appropriately applied brings considerable standardisation benefits. The Canadian Immunology Quality Assessment Program (CIQAP) detected situations where operators did not manually override automated software in the event of improper output on the Epics XL and FC500 CD4 immunophenotyping platforms. The automated gating algorithm identifies lymphocytes using a double gate strategy based on CD45 × side scatter (SS) gating and a light scatter FS × SS gate known to fail with sub optimal specimens. METHOD: To generate correct interpretation and results CIQAP introduced a simple protocol modification, bypassing the light scatter gate to include all cells characterized by the CD45 gate. Seventeen problem cases were reanalysed for both absolute and relative T-cell subsets accuracy and compared to the CIQAP group mean values. Results were found to be associated with the percentage of lymphocytes excluded by the automated light scatter gate. RESULTS: The modified manual protocol resolved poor performance in 14 instances out of 17 problem cases. It was found to improve accuracy when the light scatter gate excluded greater than 5% of the cells. The remaining three cases had a lymphocyte recovery of greater than 94.6% in the original automated analysis. CONCLUSION: There is a risk in relying solely on automated gating procedures when using the Epics XL and FC500 CD4 immunophenotyping platforms. Laboratory managers have the responsibility to intervene when required. EQA providers are equally responsible to alert the clinical laboratories of the need to update operator training to deal with stressed specimens. © 2016 International Clinical Cytometry Society.

Liver development in a rat model of fetal alcohol syndrome.

The inhibitory effects of alcohol on hepatic growth in adults raises the possibility that the liver may be involved in fetal alcohol syndrome (FAS) in infants. To test this hypothesis, pregnant Sprague-Dawley rats were fed liquid diets containing either ethanol as 36% of the total calories, or were allowed ad libitum feeding of a control liquid diet (controls) throughout pregnancy. Other dams were exposed to the ethanol diet only during the first or last half of pregnancy. Pups delivered of dams exposed to the various diets (N = 40-45/group) were killed at 1, 3, 7, and 14 days of age. In addition to brain weights, crown-rump lengths, and facial features, the following parameters of liver development were documented; liver weight, liver/body weight ratio, liver histology, hepatic ornithine decarboxylase activity (ODC), hepatic protein content, and rate of hepatic DNA synthesis (as determined by [3H]thymidine incorporation). The results revealed that pups exposed to ethanol throughout pregnancy but not ad libitum control diet pups had brain weights, crown-rump lengths, and facial features in keeping with FAS. With respect to liver development, the livers in FAS pups were consistently smaller than in the control group. However, total body weights were decreased to a greater extent, such that when corrected for body weights, the smaller livers in FAS pups only became significant on day 14 of life. Liver histology was similar in the two groups with no signs of active inflammation or fibrosis. Hepatic ODC activity was also similar, indicating no impairment in polyamine synthesis. Hepatic DNA synthesis rates were decreased in FAS pups at all time intervals. Pups delivered of dams exposed to ethanol during either the first or last half of pregnancy had results comparable to those of controls. To identify the mechanism(s) responsible for these findings, a second series of experiments was performed wherein the hepatic expression of the following factors associated with liver development were documented by northern-blot analyses; growth hormone receptor (GHr), insulin-like growth factor-I (IGF-I) and -II (IGF-II) and IGF binding proteins (IGFBPs) 1, 2, 3, and 4 mRNA on gestational days 16 and 20 and postpartum days 1 and 7. In this series, a third group of pups derived from dams in whom caloric consumption was matched to that of the ethanol-fed dams (isocaloric controls) were also studied. The results revealed no consistent differences in GHr, IGF, or IGFBP mRNA expression in the three groups. In conclusion, liver development and hepatic DNA synthesis were significantly impaired in this animal model of FAS. That impairment, however, was not associated with decreases in either polyamine synthesis or disturbances in the hepatic component of the GH/IGF/IGFBP axis.

Vaccinia virus infection disarms the mitochondrion-mediated pathway of the apoptotic cascade by modulating the permeability transition pore.

Many viruses have evolved strategies that target crucial components within the apoptotic cascade. One of the best studied is the caspase 8 inhibitor, crmA/Spi-2, encoded by members of the poxvirus family. Since many proapoptotic stimuli induce apoptosis through a mitochondrion-dependent, caspase 8-independent pathway, we hypothesized that vaccinia virus would encode a mechanism to directly modulate the mitochondrial apoptotic pathway. In support of this, we observed that Jurkat cells, which undergo Fas-mediated apoptosis exclusively through the mitochondrial route, were resistant to Fas-induced death following infection with a crmA/Spi-2-deficient strain of vaccinia virus. In addition, vaccinia virus-infected cells subjected to the proapoptotic stimulus staurosporine exhibited decreased levels of both cytochrome c released from the mitochondria and caspase 3 activation. In all cases we found that the loss of the mitochondrial membrane potential, which occurs as a result of opening the multimeric permeability transition pore complex, was prevented in vaccinia virus-infected cells. Moreover, vaccinia virus infection specifically inhibited opening of the permeability transition pore following treatment with the permeability transition pore ligand atractyloside and t-butylhydroperoxide. These studies indicate that vaccinia virus infection directly impacts the mitochondrial apoptotic cascade by influencing the permeability transition pore.

Growth status and related medical conditions among refugee children in Massachusetts, 1995-1998.

OBJECTIVES: This study analyzed growth and identified related medical conditions among refugee children in Massachusetts. METHODS: Between July 1995 and June 1998, 1825 refugee children were screened. Variables included positive tuberculin (purified protein derivative; PPD) test; dental abnormalities; pathogenic parasites; weight-for-age, height-for-age, and weight-for-height z scores; and body mass index greater than the 84th percentile. RESULTS: Of all the children, 21% had parasites, 62% had caries, and 25% had a positive PPD reaction. Twelve percent overall and 28% younger than 2 years had anemia. Eight percent had height-for-age z scores less than -2, and 6% had weight-for-age z scores greater than +2. Of those aged 1 to 9 years, 7% had weight-for-height z scores greater than +2. Weight-for-height z scores less than -2 were concentrated among Africans and East Asians (both 8%). Height-for-age z scores less than -2 were seen among African (13%), Near Eastern (19%), and East Asian (30%) children. Weight-for-height z scores greater than +2 and body mass index greater than the 84th percentile were concentrated among children from the former Yugoslavia (8% and 15%) and the former Soviet Union (8% and 14%). CONCLUSIONS: Recently arrived refugee children have significant growth abnormalities. European refugees were overweight; those from developing countries had growth retardation.

Daily multivitamins with iron to prevent anemia in infancy: a randomized clinical trial.

This study assessed the effectiveness of multivitamins (MV) with iron as prophylaxis against iron deficiency (ID) and anemia in infancy. The study was a double-blind, randomized trial at 2 urban primary care clinics. Subjects included healthy, full-term infants enrolled at their 6-month well-child visit. Parents administered MV, either with iron or without iron, by mouth daily for 3 months. At 9 months of age, 28.3% of 310 had either anemia or ID without anemia. Among infants with any adherence, anemia was found in 11.1% of the iron group and 21.7% in the noniron group (RR=0.5, 95% CI=0.3-1.0). Iron deficiency without anemia was found in 18.5% of the iron group; 14.4% of the noniron group (p=0.46). When administered daily starting at age 6 months, standard-dose multivitamins with iron appear to reduce anemia prevalence at 9 months of age.

Hunger in legal immigrants in California, Texas, and Illinois.

OBJECTIVES: This study determined the prevalence of food insecurity and hunger in low-income legal immigrants. METHODS: We conducted a cross-sectional survey of Latino and Asian legal immigrants attending urban clinics and community centers in California, Texas, and Illinois with a food security questionnaire. RESULTS: Among 630 respondents, 40% of the households were food insecure without hunger and 41% were food insecure with hunger. Independent predictors of hunger were income below federal poverty level (odds ratio [OR] = 2.72, 95% confidence interval [CI] = 1.72, 4.30), receipt of food stamps (OR = 2.53, 95% CI = 1.57, 4.09), Latino ethnicity (OR = 2.39, 95% CI = 1.49, 3.82), and poor English (OR = 1.76, 95% CI = 1.10, 2.82). CONCLUSIONS: The prevalence of hunger among low-income legal immigrants is unacceptably high. Access to food assistance programs is important for the health and well-being of this population.

Immigration legal status and use of public programs and prenatal care.

This study assessed impacts of immigrants' legal status on utilization of prenatal care and public programs by non-citizen women. Interviews and chart reviews of 171 women were conducted less than 48 hours after delivery at an urban hospital. Among non-citizens, 67% were legally documented. No differences in prenatal care adequacy by legal status were detected among non-citizens, who as a group had less health insurance, worse socioeconomics, and less program use compared to U.S. citizens. Non-citizens also were less aware of newborn's citizenship and eligibility for public programs and benefits. Non-citizens utilized fewer programs while reporting greater economic hardship than citizens; however, non-citizens selectively used programs important for pregnancy. Given changes in eligibility for federal programs and high rates of reported food insecurity and socioeconomic hardship among non-citizens, monitoring for adverse effects on utilization of prenatal care and birth outcomes is needed.

An A/G-rich motif in the rat fibroblast growth factor-2 gene confers enhancer activity on a heterologous promoter in neonatal rat cardiac myocytes.

We have cloned the rat fibroblast growth factor-2 (FGF-2) promoter region including 1058 base pairs (bp) of 5'-flanking DNA. Complete sequencing of this promoter region revealed a 74 bp domain between nucleotides-793 and-720 that was greater than 97% A/G-rich. A repeat of the sequence 5'-AGGGAGGG-3' separated by 11 bp was located at the core of this domain. A 37 bp A/G-rich oligonucleotide containing these AGGG-repeat sequences was synthesised, and tested for function on a minimal herpes simplex virus thymidine kinase (TK) promoter, fused to the firefly luciferase gene (TKp.luc), in transiently transfected neonatal rat cardiac myocytes. Promoter activity was stimulated approximately 3 fold in the presence of AGGG-repeat sequences. This effect was neither tissue or species-specific since TK promoter activity was increased approximately 11 fold in both rat and human glial tumor cells. Four specific complexes (Cl-4) were detected between neonatal rat heart nuclear proteins and the 37 bp A/G-rich oligonucleotide by gel mobility shift assay. Competition with excess unlabelled 37 bp A/G-rich oligonucleotide revealed that two complexes represented very high affinity/specificity interactions (C2 > C4) while Cl and C3 were of lower affinity. As a result, competition with up to a 25 fold molar excess of 37 bp A/G-rich oligonucleotide led to the loss of C2 and C4, and a corresponding and transient increase in the levels of Cl and C3, which themselves were reduced with more competitor oligonucleotide. The AGGG-repeat resembles the 5'-gGGGAGGG-3' sequence previously implicated in the response of the atrial natriuretic factor promoter to the alpha-adrenergic agonist, phenylephrine. Although an additional 1.5 fold increase in TK promoter activity was detected in the presence of the 37 bp A/G-rich oligonucleotide with phenylephrine treatment of transfected myocytes, this effect was not statistically significant. Furthermore, there was no difference in the gel mobility shift (Cl-4) pattern obtained with the 37 bp A/G-rich oligonucleotide and nuclear protein isolated from neonatal rat cardiac myocytes grown in the presence or absence of norepinephrine. These data suggest that the A/G rich sequences in the rat FGF-2 gene 5'-flanking DNA, including the AGGG-repeat, are able to confer stimulatory activity on a promoter in a tissue- and species-independent manner, but alone are not able to induce a significant phenylephrine response in neonatal rat cardiac myocytes.

The UDHR and the physician's role.

PIP: Upon the 50th anniversary of the Universal Declaration of Human Rights (UDHR), medical professionals should pay attention to the widening gap between the rich and the poor. The rights outlined in the UDHR include the traditional province of civil and political rights, as well as the right to a standard of living adequate for the health and well-being of an individual and his or her family, including food, clothing, housing, medical care, and necessary social services. The health profession should work to improve standards of living for those who are disenfranchised. Socioeconomic status is directly related to health status, with inequities in the distribution of wealth affecting morbidity and mortality. The global polarization of wealth and its effect upon health are best reflected in the US, the world's wealthiest country. Until societies confront the disparity of wealth both between and within countries, basic human needs will remain unmet and health for all will not be attained.^ieng

Immigration reporting laws: ethical dilemmas in pediatric practice.

Objectives. This study assessed the potential impact of immigration reporting requirements on pediatricians' referrals to child protective services.Methods.A random sample of 200 Massachusetts pediatricians were surveyed. Chi-square and logistic regression analyses were performed.Results.Asked whether potential deportation of the family would cause them to question or alter a decision to refer, 50% of the respondents said yes.Conclusions.Pediatricians, as mandated reporters of child abuse, will face ethical dilemmas if laws requiring reporting of immigration status are enacted. (Am J Public Health. 1998;88:967-968)

The beneficial effects of ciprofloxacin on survival and hepatic regenerative activity in a rat model of fulminant hepatic failure.

Recently, we reported that ciprofloxacin, an antimicrobial agent with gamma-aminobutyric acid (GABA(A)) receptor antagonist properties, significantly increases hepatic regenerative activity in animal models of alcohol-induced liver disease and cirrhosis. In the present study, we documented the effects of ciprofloxacin on survival and hepatic regeneration in a D-galactosamine (D-gal)-induced model of acute hepatic injury in rats. One hundred nineteen adult, male Sprague-Dawley rats (n = 19-20/group) were treated with intraperitoneal D-gal (total dose: 2.5 g/kg), followed by gastric gavage with either saline, ciprofloxacin (10, 50, or 100 mg/kg), norfloxacin (250 mg/kg), or intraperitoneal putrescine (300 micromol/kg), a potent hepatic growth promoter. Mortality rates were then documented over a 4-day period. An additional 45 rats (n = 15/group) received a sublethal dose of D-gal (1.0 g/kg), followed by gastric gavage with either saline or ciprofloxacin (100 mg/kg), or intraperitoneal putrescine (300 micromol/kg). In these rats, hepatic regenerative activity was documented at 12, 24, and 60 hours post-D-gal by 3H-thymidine incorporation into hepatic DNA and proliferating cell nuclear antigen (PCNA) staining. In the survival study, a dose-response effect of ciprofloxacin on survival was observed (ciprofloxacin: 10 mg/kg, 10%; 50 mg/kg, 26%; and 100 mg/kg, 35%) with the results in the 100-mg/kg-treated group being significant when compared with the 5% survival rate in saline-treated controls (P < .05). Survival figures in the norfloxacin- and putrescine-treated groups were not significantly improved (15% and 25%, respectively). In the regeneration study, compared with the D-gal + saline-treated control group, DNA synthesis rates at 60 hours were increased in the D-gal + ciprofloxacin and D-gal + putrescine groups (10.2 +/- 3.3 vs. 18.2 +/- 5.1 and 18.8 +/- 6.8 x 10(3) dpm/mg DNA respectively; P < .05). The results of PCNA staining also supported enhanced hepatic regeneration in the ciprofloxacin-treated group at 60 hours (saline, 13.4 +/- 3.7; ciprofloxacin, 47.4 +/- 7.3; and putrescine, 8.4% +/- 2.8% hepatocytes staining positive). Ciprofloxacin at a dose of 100 mg/kg significantly improves survival and hepatic regenerative activity in this animal model of acute hepatic injury.

The nutritional impact of breakfast consumption on the diets of inner-city African-American elementary school children.

To determine the contribution of breakfast-eating behavior to dietary adequacy among low-income African-American children, 1151 children attending grades two through five at four elementary schools in East Orange, New Jersey were studied. Results of a 4-day eating behavior survey and a 24-hour dietary recall reveal that on any given day, 12% to 26% of children attend school without having eaten anything. Thirty-six percent of the children were obese, which did not vary with breakfast-eating behavior. A significantly greater proportion of the children who skipped breakfast compared to those who ate breakfast failed to achieve dietary adequacy for nearly every nutrient studied. More than one third of breakfast skippers consumed < 50% of the recommended dietary allowance for vitamins A, E, B6, and folacin, and nearly one fourth consumed < 50% of the recommended dietary allowance for calories, vitamin C, calcium, and iron. Not eating breakfast results in substantial deficits in dietary intake of a variety of essential nutrients among low-income African-American school children. Efforts to improve the nutritional status of children should include nutrition education to promote breakfast.

School Breakfast Program and school performance.

To test the hypothesis that participation in the School Breakfast Program by low-income children is associated with improvements in standardized achievement test scores and in rates of absence and tardiness, children in grades 3 through 6 were studied in the Lawrence, Mass, public schools, where the School Breakfast Program was begun at the start of the second semester 1986-1987 school year. The changes in scores on a standardized achievement test and in rates of absence and tardiness before and after the implementation of the School Breakfast Program for children participating in the program were compared with those of children who also qualified but did not participate. Controlling for other factors, participation in the School Breakfast Program contributed positively to the 1987 Comprehensive Tests of Basic Skills battery total scale score and negatively to 1987 tardiness and absence rates. These findings suggest that participation in the School Breakfast Program is associated with significant improvements in academic functioning among low-income elementary school children.

The growth and development status of homeless children entering shelters in Boston.

In order to characterize the children who enter emergency shelters in Boston, we reviewed the data collected at intake interviews by the pediatric nurse practitioner visiting 10 family shelters and one hotel in Boston as part of the Boston Health Care for the Homeless Project. Families were interviewed soon after their entry into the shelter. Children were weighed and measured, and the Denver Developmental Screening Test (DDST) was administered. From November 1986 to November 1987, 133 families with 213 children were interviewed. Ninety-four percent of the children were in the care of their mothers, and 92 percent were younger than 5 years of age. Sixty-five percent of the families were black, 20 percent were white, and 11 percent were Hispanic. Eighty-nine percent of the families were receiving Aid to Families with Dependent Children benefits, 90 percent were receiving Medicaid benefits, 72 percent were receiving food stamps, and 52 percent were receiving benefits under the Special Supplement Food Program for Women, Infants, and Children. Eighty-five percent of the children were reported to have a regular source of primary pediatric care, and 23 percent were reported to have medical problems. Weight-for-age, weight-for-height, and height-for-age measurements were similar to those reported for national samples of low income children. Ten children (4.7 percent) were found to have abnormal or questionable DDST examinations.