Polymerase chain reaction--principle and application in medicine

The polymerase chain reaction, a new molecular technique, is of increasing importance in many areas of medical research and diagnosis. Within hours DNA sequences can be amplified millionfold with very high specificity, making detection and further analysis possible. Using multiple rounds of exponential amplification, even one copy of a gene of interest can be detected by agarose gel electrophoresis and other methods. Furthermore, minute amounts of DNA even from tissue damaged by embedding in paraffin can be analysed. Polymerase chain reaction methodology has already gained significance in many areas of medical research, e.g. diagnosis of inherited diseases, detection of viral DNA in clinical samples, cancer research and diagnosis, and characterization of gene expression.     

Congenital self-healing reticulohistiocytosis. Report of a case with 7-year follow-up and a review of the literature

A case of congenital self-healing reticulohistiocytosis in an otherwise healthy newborn boy is presented. Histological, immunohistochemical, and ultrastructural findings are described and the nosologic position of this entity is discussed.     

Protective effect of tissue ferritins in experimental Escherichia coli infection of mice in vivo.

The effect of ferritins from horse (FH) and bovine (FB) spleen and murine liver (FM) on the survival rate of CFW mice lethally infected with Escherichia coli (strain 8440-78 K 80/B) was evaluated. Ferritins given intravenously 24 h before intravenous inoculation of bacteria, protected mice most effectively from death due to infection. The effect was dose dependent. At 500 micrograms of ferritin per mouse, the maximum survival rates were 86% (FH), 81% (FM) and 79% (FB), while only 5% of the control mice survived up to the 30th day. The survival rates of animals injected with bovine serum albumin (BSA) and heat-inactivated FB were 8 and 25%, respectively. Intraperitoneal injection of FB was as effective as intravenous in enhancing the resistance of mice against bacteria. These data provide evidence for the beneficial role of tissue ferritins in nonspecific antibacterial resistance.     

Protein quality and quantity influence free amino acid levels in the brain and serum of rats during lactation

Diets containing 11 or 21% protein supplied by wheat, wheat + lysine + threonine or casein + methionine were fed to pregnant rats from conception until d 4 or 8 of lactation. Dams then were decapitated, serum was collected and brains and left inguinal-abdominal mammary glands were quickly excised and weighed. Serum and brains were frozen in liquid nitrogen and stored at -70 degrees C. Free amino acids in either serum or brain were not significantly different on d 8 as compared with d 4 of lactation. Increasing dietary protein quality or quantity increased concentrations of most essential amino acids in serum and brain. Changes in the concentrations of nonessential amino acids in serum or brain associated with increases in protein quality or quantity were variable. For all 16 amino acids measured, brain free amino acids were highly correlated with serum levels. In this experiment, pup growth varied from 0.3 g/d for the offspring of dams fed 11% wheat to 1.4 g/d for the offspring of dams fed 21% wheat + lysine + threonine. Concentrations of lysine and threonine in the brain were more than twice as high under the latter as compared with the former condition, but differences in all other amino acids among the dietary groups were small, ranging from -20% for glycine to +21% for leucine and isoleucine.     

Human calcium absorption from whole-wheat products

Fractional calcium absorption from wheat products and the influence of co-ingested wheat products on calcium absorption from milk were measured in a series of randomized crossover studies in healthy adult women. The wheat had been intrinsically labeled with 45Ca during growth. In the first study, fractional calcium absorption from leavened whole-wheat bread averaged 0.817 +/- 0.124. By comparison, absorption from milk, ingested at a comparable load in the same women, averaged only 0.589 +/- 0.111. When labeled bread was co-ingested with milk, at the same aggregate load as for bread alone, bread calcium absorption fell to 0.748 +/- 0.103 (P less than 0.05). In a second study, calcium absorption from an extruded cereal prepared from intrinsically labeled wheat bran was compared with milk. Calcium absorption from the cereal (0.223 +/- 0.046) was significantly less than from milk (0.375 +/- 0.072) (P less than 0.001). When the two were co-fed at the same total load, milk calcium absorption fell to 0.258 +/- 0.055 (P less than 0.001). In a third study, the effect of phytate hydrolysis through yeast fermentation and of Maillard browning on calcium absorption was investigated using leavened bread and underbaked and overbaked cookies, each made with intrinsically labeled wheat flour. Calcium absorption from cookies was not affected by the extent of browning and averaged 0.652 +/- 0.087. However, calcium absorption from bread in these same women averaged 0.703 +/- 0.108. This was significantly more than from the cookies (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum 25-hydroxyvitamin D concentrations of free-living subjects consuming olestra

The effect of olestra on vitamin D status was assessed in a 6-wk, double-blind, placebo-controlled study involving 202 free-living adults. Subjects consumed a total of 20 g/d of olestra or triglycerides in cookies eaten at each meal. A 20-micrograms ergocalciferol capsule was taken with each morning meal. Serum 25-hydroxyergocalciferol (25-OHD2) concentrations rose from approximately 5.7 to 39.0 and 31.7 nmol/L in the placebo and olestra groups, respectively, at week 6. At week 6, 25-OHD2 contributed 46-54% to total serum 25-OHD concentration compared with 11% at baseline. The 19% decrease in serum 25-OHD2 concentrations produced by olestra in this study is equivalent to a decrease of approximately 1.2 nmol/L under nonsupplemented dietary conditions. Ingesting 20 g olestra/d in the diet is thus not expected to affect vitamin D nutritional status.     

Use of the deoxyuridine suppression test to evaluate localized folate deficiency in rat colonic epithelium

In this study the deoxyuridine suppression test (dUST) was performed on isolated rat colonocytes to establish its value as an indicator of folate status in the colonic epithelium. [3H]thymidine incorporation into DNA was suppressed greater than 90% by deoxyuridine (dU) concentrations greater than 2.5 mumol/L. Preincubation of cells with 5-fluorouracil (1-100 mumol/L) but not methotrexate (10-100 mumol/L) resulted in a significant decrease in the degree of suppression. The dUST performed on colonocytes from folate-deficient animals displayed less suppression than on colonocytes from folate-replete animals (P less than 0.05). The abnormal degree of suppression was corrected by adding 100 mumol folinic acid/L. There was a negative correlation between the degree of suppression and the folate concentration of the colonic epithelium (P less than 0.001). These data indicate that the dUST is useful for detecting folate deficiency in the colonic epithelium and may therefore be valuable in assessing a deficiency state localized to that epithelium.     

Vitamin K in colostrum and mature human milk over the lactation period--a cross-sectional study

Vitamin K was quantitated in the milk of four groups of 15 mothers from 1 d to 6 mo postpartum in a cross-sectional study. Concentrations were 7.52 +/- 5.90 and 6.36 +/- 5.32 nmol/L (3.39 +/- 2.66 and 2.87 +/- 2.40 micrograms/L) in colostrum and mature milk, respectively. Differences between colostrum and mature milk or among samples of mature milk collected at 1, 3, and 6 mo were not statistically significant. Because of significantly increased volumes of milk over the lactation period, approximately twice as much vitamin K was delivered in mature milk as in colostrum. Within normal ranges, concentrations of vitamin K in milk were not predicted by dietary intake of vegetables or fat. Vitamin K was correlated with fat in colostrum and was localized in the lipid core of the milk fat globule but was not associated with membranes. Vitamin K in human milk is insufficient to meet recommended intakes for infants aged less than 6 mo. Population and clinical studies are needed to assess the vitamin K status of exclusively breast-fed infants and to evaluate current recommendations.