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Biomedical subjects

D Martin

Publications and source records attributed to D Martin.

At least 181 records · Page 10Linked to original sources

Characterization of Plasmodium falciparum CDP-diacylglycerol synthase, a proteolytically cleaved enzyme.

Cytidine diphosphate-diacylglycerol (CDP-DAG), an obligatory intermediate compound in the biosynthesis of the major anionic and zwitterionic phospholipids, is synthesized by CDP-DAG synthase (CDS). The gene encoding CDS was isolated from the human malaria parasite Plasmodium falciparum, based on sequence conservation to CDS from other organisms. The P. falciparum gene is located as a single copy on chromosome 14. The open reading frame (ORF) of PfCDS gene encodes a putative protein of 667 amino acids and 78 kDa. Only the C-terminal 422 amino acids share 40% homology with eukaryotic CDSs. The very long and non-conserved N-terminal region of 245 amino acids is hydrophilic and contains asparagine-rich and repetitive sequences. Two mRNA of 3.5 and 4 kb were detected. Transcription is developmentally regulated during the asexual intraerythrocytic cycle, being the weakest in the ring-stage. PfCDS enzyme activities in infected erythrocytes correlates with the transcription pattern, consistent with an increased synthesis of phospholipids in trophozoites and schizonts. Antisera raised against two synthetic peptides from the C-terminal region of PfCDS detected a single protein of 51 kDa in Western blot analysis, specific for parasitized erythrocytes. A protein of 28 kDa was recognized by an antiserum against an N-terminal peptide, indicating that PfCDS is proteolytically processed. Expression of 51- and 28-kDa proteins was developmentally regulated similar to regulation of the transcripts and the enzyme activity. The conserved C-terminal region of PfCDS, cloned into a eukaryote expression vector and transfected in COS-7 cells, showed a two-fold increase CDP-DAG synthase activities, indicating that the isolated gene most likely encoded the P. falciparum CDS enzyme.

Amino Acid Sequence↗

The expression of teneurin-4 in the avian embryo.

The teneurins are a family of four transmembrane proteins expressed in the developing vertebrate nervous system, though the Drosophila teneurin ten-m is also a pair-rule gene. Whole-mount in situ hybridization was used to localize teneurin-4 transcripts in the chicken embryo. The earliest signal is detected at stage 19 in the somites and limb buds. By stage 20 teneurin-4 transcripts are detected in temporal periocular mesenchyme, branchial arches, diencephalon and somites. Teneurin-4 expression in the limbs changes dramatically during development. Between stages 19 and 21 teneurin-4 expression is concentrated proximally in the zone of polarizing activity. Between stages 24 and 26 teneurin-4 is expressed in the mesenchyme of the anterodistal part of the limb. As in Drosophila, vertebrate teneurins are expressed not only in the nervous system, but also in non-neuronal tissues during pattern formation.

Amino Acid Sequence↗

Nucleosides and nucleotides: natural bioactive substances in milk and colostrum.

Nucleotides, nucleosides and nucleobases belong to the non-protein-nitrogen (NPN) fraction of milk. The largest amounts of ribonucleosides and ribonucleotides--ribose forms only were considered in this review--were measured directly after parturition in bovine milk and other ruminants as well as in the milk of humans. Generally, concentrations of most of the nucleos(t)ides tend to decrease gradually with advancing lactation period or nursing time. The species-specific pattern of these minor constituents in milk from different mammals is a remarkable property and confirms, at least, the specific physiological impact of these minor compounds in early life. The physiological capacity of these compounds in milk is given by the total potentially available nucleosides. The main dietary sources of nucleos(t)ides are nucleoproteins and nucleic acids which are converted in the course of intestinal digestion into nucleosides and nucleobases the preferred forms for absorption in the intestine. Thus, nucleosides and nucleobases are suggested to be the acting components of dietary and/or supplemented nucleic acid-related compounds in the gut. They are used by the body as exogenous trophochemical sources and can be important for optimal metabolic functions. Up to 15 % of the total daily need for a breast-fed infant was calculated to come from this dietary source. Concerning their biological role they not only act as metabolites but are also involved as bioactive substances in the regulation of body functions. Dietary nucleotides affect immune modulation, e.g. they enhance antibody responses of infants as shown by a study with more than 300 full-term healthy infants. Dietary nucleos(t)ides are found to contribute to iron absorption in the gut and to influence desaturation and elongation rates in fatty acid synthesis, in particular long-chain polyunsaturated fatty acids in early stages of life. The in vitro modulation of cell proliferation and apoptosis has been described by ribonucleosides, in particular by modified components using human cell culture models. Due to the bio- and trophochemical properties of dietary nucleos(t)ides, the European Commission has allowed the use of supplementation with specific ribonucleotides in the manufacture of infant and follow-on formula. From the technochemical point of view, the ribonucleoside pattern is influenced by thermal treatment of milk. In addition ribonucleosides are useful indicators for quantifying adulterations of milk and milk products.

Animals↗

Carriage of N. lactamica in a population at high risk of meningococcal disease.

Carriage of Neisseria lactamica among household contacts of meningococcal disease (MCD) cases was investigated during an epidemic in Auckland, New Zealand. The overall carriage rate for N. lactamica was 10.5% (95% CI 7.4-13.5%) with a peak carriage rate in 2-year-olds of 61.5% (95% CI 26.6-88.1%). Factors associated with a significant (P < 0.05) increase in the likelihood of carriage included runny nose, the number of people per bedroom and youth. Genetic analysis of isolates revealed a striking correlation of strains within the same household but a high level of diversity between households, suggesting that household contact is an important factor in acquisition. For household contacts aged less than 5 years, there was a higher rate of carriage amongst those in contact with MCD cases under 8 years old than for contacts of cases aged 8 years and over. It is likely that development of MCD is a reflection of the nature and intensity of the exposure to a virulent strain of N. meningitidis, coupled with an absence of host resistance among those individuals not carrying N. lactamica.

Adolescent↗

Multi-variate analysis predicts clinical outcome 30 days after middle cerebral artery infarction.

BACKGROUND AND PURPOSE: To evaluate the functional prognostic value of proton magnetic resonance spectroscopy performed within the 5 days of an infarction of the middle cerebral artery territory, compared with previously demonstrated prognostic factors. METHODS: Proton magnetic resonance spectroscopy was performed on 77 consecutive non-comatosed patients during the acute stage of middle cerebral artery infarction. The functional status was determined for each patient via the Orgogozo score. Proton magnetic resonance spectroscopic data were acquired in the infarction and in contra-lateral normal tissue and the results were expressed as metabolite ratios. Correlations were evaluated between the Orgogozo score at day 1 and day 30, the age, the sex, the volume of the infarction, and the metabolic ratios. RESULTS: In a monovariate analysis, the decrease of the NAA/choline ratio was correlated with a low Orgogozo score at days 1 and 30 (P<0.05) and with a large infarction (P<0.05). A stepwise analysis showed a significant relationship between the Orgogozo score at day 30 and the Orgogozo score at day 1, the sex, the volume of infarction, and the NAA/Cho ratio within the infarction. CONCLUSIONS: Our work demonstrates that a good clinical outcome at day 30 depends on a good initial clinical score at day 1, a small volume of infarction, a small decrease of NAA/Cho, and being of the female gender.

Adult↗

Effects of intrapartum administration of invert sugar and D5LR on neonatal blood glucose levels.

OBJECTIVE: To compare the effects of D5LR and invert sugar administered intrapartum on neonatal blood glucose concentrations. STUDY DESIGN: This is a prospective, randomized, double-blind study. A total of 32 insulin-requiring diabetic patients were randomized to receive either intravenous 10% invert sugar or lactated Ringer's solution with 5% dextrose (D5LR). Regular insulin was given intravenously with an infusion pump to maintain the plasma glucose concentration between 60 and 90 mg/dl. Neonatal blood glucoses were measured at 30 minutes after birth, four times every hour, and thereafter as indicated. Student's t-test was used for continuous variables and Fisher's exact test was used for categorical data. RESULTS: There were no differences in neonatal blood glucose levels, incidence of neonatal hypoglycemia, or length of neonatal hospital stay between the two groups. CONCLUSION: Intrapartum administration of invert sugar is not associated with better neonatal homeostasis of glucose compared with using D5LR. Thus, given the expense of invert sugar, D5LR appears to be preferable for intrapartum control of maternal blood glucose.

Adult↗

Tight junction proteins ZO-1, ZO-2, and occludin along isolated renal tubules.

BACKGROUND: Tight junctions play a critical role in tubular function. In mammalian kidney, the transepithelial electrical resistance and the complexity of the tight junction increase from the proximal to the collecting tubule. The differential expression of three tight junction proteins, ZO-1, ZO-2, and occludin, along isolated rabbit renal tubules is examined in this article. METHODS: Microdissected rabbit renal tubules were processed for immunofluorescence detection of ZO-1, ZO-2, and occludin. The quantitation of these proteins was done by Western blot determinations in Percoll isolated tubules. RESULTS: ZO-1 stained cell boundaries independently of the identity of the tubule. However, the amount found in distal segments was significantly higher than that expressed in proximal regions. ZO-2 in the proximal region was found diffusely distributed in the cytoplasm, with faint staining at cell borders, while a clear signal at cell perimeters was detectable from the Henle's loop to collecting tubules. Nuclear staining of ZO-2 was found along the whole nephron. The presence of occludin at the proximal region was faint and discontinuous, while its expression in the more distant portions was conspicuous. The quantity of ZO-2 and occludin present at the distal region was significantly higher compared with the proximal segment. CONCLUSIONS: The distribution of ZO-1, ZO-2, and occludin follows the increase in junction complexity encountered in renal tubules. The amount of the three proteins found in proximal and distal segments is significantly higher in the latter.

Animals↗

A novel mechanism for skeletal resistance in uremia.

BACKGROUND: In treating secondary hyperparathyroidism, the target level of serum intact parathyroid hormone (I-PTH) should be three to five times normal to prevent adynamic bone disease. In circulation, there is a non-(1-84) PTH-truncated fragment, likely 7-84, which, in addition to PTH 1-84, is measured by most I-PTH immunoradiometric (IRMA) assays, giving erroneously high I-PTH values. We have developed a new IRMA assay in which the labeled antibody recognizes only the first six amino acids of the PTH molecule. Thus, this new IRMA assay (Whole PTH) measures only the biologically active 1-84 PTH molecule. METHODS: Using this new IRMA assay (Whole PTH) and the Nichols "intact" PTH assay, we compared the ability of each assay to recognize human PTH (hPTH) 1-84 and hPTH 7-84 and examined the percentage of non-1-84 PTH in circulation and in parathyroid glands. Possible antagonistic effects of the 7-84 PTH fragment on the biological activity of 1-84 PTH in rats were also tested. RESULTS: In 28 uremic patients, PTH values measured with the Nichols assay, representing a combined measurement of both hPTH 1-84 and hPTH 7-84, were 34% higher than with the Whole assay (hPTH 1-84 only); the median PTH was 523 versus 318 pg/mL (P < 0.001). Similar results were found in 14 renal transplant patients. In osteoblast-like cells, ROS 17.2, 1-84 PTH (10-8 mol/L) increased cAMP from 18.1 +/- 1.25 to 738 +/- 4.13 mmol/well. Conversely, the same concentration of 7-84 PTH had no effect. In parathyroidectomized rats fed a calcium-deficient diet, 7-84 PTH was not only biologically inactive, but had antagonistic effects on 1-84 PTH in bone. Plasma calcium was increased (0.65 mg/dL) two hours after 1-84 PTH treatment, while 7-84 PTH had no effect. When 1-84 PTH and 7-84 PTH were given simultaneously in a 1:1 molar ratio, the calcemic response to 1-84 PTH was decreased by 94%. In normal rats, the administration of 1-84 PTH increased renal fractional excretion of phosphate (11.9 to 27.7%, P < 0.001). However, when 1-84 PTH and 7-84 PTH were given simultaneously, the 7-84 PTH decreased the phosphaturic response by 50.2% (P < 0.005). Finally, in surgically excised parathyroid glands from six uremic patients, we found that 44.1% of the total intracellular PTH was the non-PTH (1-84), most likely PTH 7-84. CONCLUSION: In patients with chronic renal failure, the presence of high circulating levels of non-1-84 PTH fragments (most likely 7-84 PTH) detected by the "intact" assay and the antagonistic effects of 7-84 PTH on the biological activity of 1-84 PTH explain the need of higher levels of "intact" PTH to prevent adynamic bone disease.

Animals↗

Expression of endothelial nitric oxide synthase by extravillous trophoblast cells in the human placenta.

Previous reports have documented the expression of endothelial nitric oxide synthase (eNOS) expression by the syncytiotrophoblast layer of the villus in the human placenta. In contrast, the underlying villous cytotrophoblast cells do not express the enzyme. Because extravillous cytotrophoblasts have not been as extensively investigated, our objective was to test whether these cells express eNOS. Using both a mouse monoclonal and a rabbit polyclonal antibody, we demonstrated immunoreactive eNOS in trophoblast cell columns emanating from anchoring villi in second trimester placentae. Cytokeratin positive trophoblast cells lying beneath remnant anchoring villi, lining decidual blood vessels and scattered throughout the basal plate of normal term and pre-eclamptic placentae also expressed immunoreactive eNOS. By Western analysis, the monoclonal and polyclonal antibodies were shown to be absolutely and relatively specific for eNOS, respectively. The finding of immunoreactive eNOS expression by extravillous trophoblast cells was substantiated by in situ hybridization. Using riboprobes generated from a bovine eNOS cDNA, we demonstrated specific hybridization in the endothelium of blood vessels in the umbilical cord, thus validating the in situ hybridization methodology, as well as specific hybridization in the extravillous trophoblast cells of the basal plate in normal term placenta. In conclusion, several different populations of extravillous trophoblast cells in the basal plate of the human placenta express eNOS.

Animals↗

The influence of microwave heating on the characteristics of polyelectrolytes.

Results obtained by microwave (MW) and simultaneous microwave and electron beam (MW + EB) application in the chemistry of acrylamide and acrylic acid co-polymers (PA type polyelectrolytes) are presented. Comparative results concerning the molecular weight (Mw) and Huggins' constant for the acrylamide copolymers obtained by classical heating, MW heating, EB irradiation and simultaneous MW + EB treatment are reported. MW heating produces high PA water solubility but median Mw values; EB irradiation gives high Mw values but associated with a cross-linked structure (poor water solubility) while MW energy addition to EB energy gives simultaneously high Mw values and high PA water solubility. A MW installation of 2.45 GHz and 2.5 kW, designed to provide small-scale commercial production of PA polyelectrolytes, is described.

Journal Article↗

RDP: detection of recombination amongst aligned sequences.

SUMMARY: Recombination Detection Program (RDP) is a program that applies a pairwise scanning approach to the detection of recombination amongst a group of aligned DNA sequences. The software runs under Windows95 and combines highly automated screening of large numbers of sequences with a highly interactive interface for examining the results of the analyses.

Computational Biology↗

Logistics and technique for procurement of intestinal, pancreatic, and hepatic grafts from the same donor.

OBJECTIVE: To assess a technique for simultaneous recovery of the intestine, pancreas, and liver from the same donor. SUMMARY BACKGROUND DATA: With the more frequent use of pancreatic and intestinal transplantation, a procurement procedure is needed that permits retrieval of both organs as well as the liver from the same cadaveric donor for transplantation to different recipients. It is believed by many procurement officers and surgeons, however, that this objective is not technically feasible. METHODS: A technique for simultaneous recovery of the intestine, pancreas, and liver was used in 13 multiorgan cadaver donors during a 26-month period, with transplantation of the organs to 33 recipients. The intestine was removed from 11 donors separately and in continuity with the pancreas in the other 2. Six additional pancreases were excised and transplanted separately. Thirteen livers were retrieved, one of which was discarded because of steatorrhea. Ten of the remaining 12 livers were transplanted intact; the other 2 were split in situ and used as reduced-size hepatic allografts in four recipients. RESULTS: None of the 11 intestinal, 6 pancreatic, 2 intestinal-pancreatic, or 14 whole or partial liver allografts sustained serious ischemic injury or were lost as a result of technical complications. One liver recipient died 25 months after surgery of recurrent C virus hepatitis. The other 32 recipients had adequate allograft function with a mean follow-up of 8 months. CONCLUSION: It was possible using the described technique to retrieve intestine, pancreas, and liver allografts safely from the same donor and to transplant these organs to different recipients.

Adolescent↗

Effects of blast exposure on exercise performance in sheep.

BACKGROUND: The effects of blast on maximal exercise performance were investigated in sheep that were trained to perform maximal exercise. METHODS AND RESULTS: Sheep were fully instrumented for determination of pulmonary and systemic hemodynamics. Blast exposure was administered by using a compressed air driven shock tube that was positioned to primarily produce cardiopulmonary injury. Four levels of exposure were used that were known to produce sublethal injury ranging from little or no grossly observable cardiopulmonary injury (level 1) to confluent ecchymosis of the heart, lung, or both (level 4). We evaluated maximal exercise performance 1 hour after exposure to level 1, level 2, and level 3 and 24 hours after level 3 and level 4. VO2max was not significantly decreased 1 hour after exposure to level 1 but was decreased after exposure to level 2 (29.9%) and level 3 (49.3%). Significant improvement in exercise performance was observed in 24 hours, as VO2max was not significantly decreased 24 hour after level 3. VO2max was decreased 24 hour after level 4 injury (30.8%). CONCLUSION: Cardiovascular data collected during exercise suggested that acute cardiopulmonary injury is responsible for the exercise performance decrement observed 1 hour after exposure and that significant recovery of function is observed 24 hours after blast injury.

Animals↗

Effect of oral creatine supplementation on isokinetic torque production.

PURPOSE: This study was conducted to examine the effect of oral creatine supplementation on the decline in peak isokinetic torque of the quadriceps muscle group during an endurance test. METHODS: Twenty-three active, but untrained, male subjects performed isokinetic strength tests on a Cybex II dynamometer at 180 degrees x s(-1). The protocol consisted of pre- and post-tests with five sets of 30 maximum volitional contractions with a 1-min rest period between sets. Subjects returned to perform the posttest after 5 d of placebo (4 x 6 g glucose x d(-1), N = 12) or creatine (4 x 5 g creatine + 1 g glucose x d(-1), N = 11) supplementation. Supplements and testing were administered in a double blind fashion. Peak torque was measured during each contraction and the 30 contractions were averaged for each set. RESULTS: A three-way mixed ANOVA with one between factor (placebo vs creatine) and two within factors (pre/post supplementation and sets 1-5) revealed no significant interactions, P > 0.05. The placebo vs creatine main effect was also nonsignificant, whereas the pre/post and set effects were significant (P < 0.05). Peak torque increased (approximately 3%) from pre- to post-testing, (P = 0.04), but the absolute magnitude of the differences is unlikely to be of any practical significance. Peak torque decreased from sets 1 to 4, whereas sets 4 and 5 were not different. A priori contrasts comparing the creatine group's performance pre vs post test for the fourth and fifth sets were nonsignificant (P > 0.05). CONCLUSIONS: Based on within and between group comparisons, we were unable to detect an ergogenic effect of oral creatine supplementation on the decline in peak torque during isokinetic exercise at 180 degrees x s(-1).

Adult↗

Soleus-fibula free transfer in lower limb reconstruction.

Free-fibula transfer has been widely used since 1975. Many modifications have been described; one of them, association of the lateral part of the soleus muscle to the fibula, is reported here through a 14-case series. This composite flap is intended for extensive defects of the lower limbs involving bone and soft tissues. The flap is considered by the authors to be reliable, with a constant vascularization. A 20-cm length offibula may be harvested associated either with the lateral part of the soleus muscle or with the whole muscle. Moreover, the soleus muscle represents a vascular security inasmuch as it preserves both medullar and periosteal bone supply. Fourteen cases have been performed by the authors since 1978 and could be reviewed with a minimum 2-year follow-up. Average length of bone defect was 12 cm, and average length offibula harvested was 18.6 cm. Soft-tissue defect was always associated and ranged from 8 x 4 cm to 20 x 30 cm. The fibula was harvested with the lateral part of the soleus muscle in 10 cases and with the whole soleus muscle in 4 cases. One total treatment failure was reported and was related to intimal degenerative lesions on veins used for arteriovenous bypass. In other patients, mean time for bone healing was 11 months. Patients could walk again, on average, 17 months after reconstruction. Sequelae at the donor site were minimal.

Adolescent↗