Factors influencing the glomerular filtration rate increase after cadaveric renal transplantation: a multivariate analysis.
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Cadaver↗
Biomedical subjects
M Gallo
Publications and source records attributed to M Gallo.
Implications on human fertility of the 677C-->T and 1298A-->C polymorphisms of the MTHFR gene: consequences of a possible genetic selection.
Mutant alleles with the 677C-->T and 1298A-->C polymorphisms of the MTHFR gene, and consequent lower methylentetrahydrofolate reductase enzyme activity, have been related to higher plasma homocysteine levels, which are associated with cardiovascular diseases. We assessed the genotype frequencies, degrees of fertility and homocysteine levels, and discuss a possible genetic selection for the gene polymorphisms studied. A total of 1777 subjects (897 women and 880 men), divided into four age groups, were genotyped by PCR and restriction fragment length polymorphism. The total homocysteine concentration in plasma was determined by fluorescence polarization immunoassay. Based on random pairs and linkage disequilibrium of the two polymorphisms, we estimated the rate of fetal non-viability according to the combinations of these two polymorphisms to be 4.63% for the group >24 years old and 6.31% for the group <24 years old. We detected an increased frequency of mutant alleles in the youngest age group, coincident with a generally increased folate intake by pregnant women in Spain. The genetic selection detected leads to an increase in mutated individuals, the number of whom could increase four-fold over the next 75 years. Although generally reduced in the younger age groups, the homocysteine plasma levels were shown to increase in individuals according to the number of mutations, especially those of the 677T allele.
Adult↗
Use of CMFDA and CMTMR fluorescent dyes in FACS-based antibody screening.
Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a control cell line not expressing the antigen. Antibodies binding only to the transfected cell line are then selected for further analysis. This approach can be challenging if a large number of antibodies need to be screened and the antibody quantities are limited. Here we describe a novel method that combines the above two steps of FACS screening into a single step, based on the use of two fluorochromes, CMFDA and CMTMR, to stain transfected and control cell lines, respectively. Antibodies conjugated to a thirdfluorochrome are then added to the combined cells, followed by three-color FACS analysis. The newly modified FACS method is simple, sensitive, and high throughput. It can be used for antibody screening in multiple cell lines simultaneously.
Animals↗
Taste aversion learning and aging: a comparison with the effect of dorsal hippocampal lesions in rats.
The relationship between hippocampal function and aging was explored in Wistar rats using taste aversion learning by comparing the performance of adult dorsal hippocampal lesioned and fifteen-month-old intact rats with that of adult intact rats. In experiment 1 the conditioned blocking phenomenon was absent in the hippocampal and the aging rats. Unlike the adult intact rats, the hippocampal and aging rats were not impaired in acquiring a learned aversion to a cider vinegar solution (3 %) presented as a serial compound with a previously conditioned saccharin solution (0.1 %). In experiment 2 both the hippocampal and the aging rats developed reduced aversions to a saline solution (0.5 %) followed by an i.p. injection of lithium chloride (0.15 M; 2 % b.w.) if the taste solution was previously preexposed without consequences. This latent inhibition effect was similar to that seen in intact adult rats. In both experiments, the aging rats exhibited enhanced conventional learned taste aversions. It is concluded that aging is not a unitary process but induces both hippocampal dependent and hippocampal independent complex changes in the functioning of the neural circuits, implementing taste aversion learning.
Acetic Acid↗
Effects of framing on teratogenic risk perception in pregnant women.
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Abnormalities, Drug-Induced↗
Conditioned blocking is re-established by neurotransplantation in mature rats.
Aging has been associated with a decay of hippocampal function that may begin well before senescence. Conditioned blocking is a complex learning phenomenon that requires an intact hippocampus in young-adult rats and is absent in middle-aged rats. The aim of the present study was to test the possibility of re-establishing conditioned blocking in 17-month-old Wistar rats by neurotransplantation. Solid embryonic hippocampal or nigral tissue was bilaterally transplanted in the proximity of the dorsal hippocampus (lateral ventricle and alveus). Conditioned blocking of an aversion to a cider vinegar (3%) solution presented in compound with a previously conditioned saccharin solution (0.1%) appeared 14 days after transplantation and persisted 3 months later only in the hippocampal grafted group, showing the possibility of restoring age-related cognitive deficits.
Acetic Acid↗
[Effects of levofolinic acid on plasma homocysteine concentrations in healthy and young women in preconceptional care].
BACKGROUND: Increases in total plasmatic homocysteine (tHcy) represents a risk factor for neural tube defects. We studied the effects of levofolinic acid (l,5-formyl-tetrahydrofolic) on the plasmatic tHcylevels in women of child-bearing age. MATERIALS AND METHOD: Healthy women aged 18-35 years (n = 30) received levofolinic acid, 5 mg/day,orally for 30 days. Both tHcy and intraerythrocytic folate levels were measured before treatment (day 0), on days 2, 5, 10 and 30 within the treatment period and on days 30 (day 60) and 60 (day 90) after the treatment was finished. Plasmatic tHcy was measured by fluorescence polarisation immunoassay and intraerythrocyticfolates by chemiluminescent immunoassay. RESULTS: Plasmatic tHcy decreased from the second day of treatment onwards (day 0 vs. 2: mean of difference: -1.24 micromol/l; CI 95% = -0.84 to -1.63; p < 0.001). The maximum decline (32.3%) was observed after 30 days (mean of difference = -2.72 micromol/l; CI 95% = -2.20 to -3.24; p < 0.001).After finishing the treatment, the hypohomocysteinic effect persisted up to days 60 (mean of difference = -2.67 micromol/l; CI 95% = -2.07 to -3.26; p < 0.001)and 90 (mean of difference = -1.49 micromol/l; CI 95% = -0.94 to -2.03; p < 0.001). The response was greater when the plasmatic tHcy concentration was >= 9 micromol/l. CONCLUSIONS: Levofolinic acid leads to a earlier, intense and persistent drop of the plasmatictHcy levels.
Adult↗
Generation of high titer antisera in rabbits by DNA immunization.
Mesothelin is a GPI-linked, membrane-associated differentiation antigen that is over-expressed in several forms of human cancers. Intradermal injection into rabbits of plasmid DNA encoding full length mesothelin resulted in antisera with titers as high as 1:100,000. Each immunization consisted of 320 microg of DNA delivered into 4 sites. After the initial three injections antisera titers were moderate (between 10 to 30,000) and fell over the course of about 7 weeks. When the titers had fallen, an injection of a booster dose of DNA resulted in very high titers of antisera. These antisera contained IgGs that could bind to both recombinant mesothelin made in Eschericha coli and to mesothelin present on human cells in Western blots and in immunofluorescence assays. These observations indicate that simple intradermal DNA immunization of rabbits can result in high titers of antibodies that can be used for a variety of purposes.
Animals↗
Analysis of bacterial lipodepsipeptides by matrix-assisted laser desorption/ionisation time-of-flight and high-performance liquid chromatography with electrospray mass spectrometry.
Strains of certain plant pathogenic bacteria, in particular several pathovars of Pseudomonas syringae, are known to produce cyclic lipodepsipeptides (LDPs) endowed with peculiar structural features and noticeable biological activities. In this study, a mass spectrometry procedure is proposed for screening LDP-producing bacterial strains and for identifying and assessing individual LDPs. After matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) screening of thirteen P. syringae strains for LDP production, the extracts from culture filtrates of eight positive strains were subjected to electrospray mass spectrometry for the identification of LDPs. Five strains were found to produce two forms of syringomycins (SR-E and SR-G) and two forms of syringopeptin 25 (SP25A and SP25B); two strains produced SR-E, SR-G and a new form of SP22; one strain produced syringotoxin (ST) and syringostatin A (SS-A) in addition to SP25A and SP25B. The yield in culture of two major LPDs: SR-G (3.2-13.8 mg x L(-1)) and SP25A (41.6-231.5 mg x L(-1)) was assessed by and high-performance liquid chromatography with electrospray mass spectrometry (HPLC/ESI-MS) in both scan and single ion monitoring (SIM) modes. Results of this investigation showed that the mass spectrometry protocol developed here is a precise and reliable method for screening bacterial strains for LDP production and for assessing the amount of each metabolite under various culture conditions. This could be of practical value in view of potential applications, e.g. biocontrol of post-harvest fungal diseases.
Amino Acid Sequence↗
Can herbal products be used safely during pregnancy? Focus on echinacea.
QUESTION: Many of my patients are now using herbal medicines; some even use them during pregnancy. As we enter the "cold and flu" season, many are inquiring about use of the herb echinacea to prevent these ailments. Is there any evidence to suggest that use of echinacea during pregnancy is safe? ANSWER: Although herbal products have been used in the past during pregnancy and delivery, there is little evidence showing they are safe. Many authoritative reviews of echinacea report that its safety for use during pregnancy has not been established. A recent Motherisk study showed that use of echinacea during the first trimester of pregnancy was not associated with increased risk of major malformations.
Adult↗
Pregnancy outcome following gestational exposure to echinacea: a prospective controlled study.
BACKGROUND: Echinacea products are among the most popular phytomedicines on the North American market. Since at least half of all pregnancies are unplanned, many women inadvertently use echinacea in their first trimester. Presently, there is a paucity of information regarding the gestational safety of this herb. The primary objective of this study was to evaluate the safety of echinacea in pregnancy when used for upper respiratory tract ailments. PATIENTS AND METHODS: The study group consisted of women who were prospectively followed up after contacting the Motherisk Program regarding the gestational use of echinacea. This cohort was disease-matched to women exposed to nonteratogenic agents by maternal age, alcohol, and cigarette use. Rates of major and minor malformations between the groups were compared. RESULTS: A total of 206 women were enrolled in the study group after using echinacea products during pregnancy; 112 women used the herb in the first trimester. There were a total of 195 live births, including 3 sets of twins, 13 spontaneous abortions, and 1 therapeutic abortion. Six major malformations were reported, including 1 chromosomal abnormality, and 4 of these malformations occurred with echinacea exposure in the first trimester. In the control group, there were 206 women with 198 live births, 7 spontaneous abortions, and 1 therapeutic abortion. Seven major malformations were reported. There were no statistical differences between the study and control groups for any of the end points analyzed. CONCLUSIONS: This first prospective study suggests that gestational use of echinacea during organogenesis is not associated with an increased risk for major malformations.
Congenital Abnormalities↗
Bilateral tetrodotoxin blockade of the rat vestibular nuclei substitutes the natural unconditioned stimulus in taste aversion learning.
The aversive effects of bilateral transient blockade of the lateral vestibular nucleus caused by tetrodotoxin microinjections were tested using conditioned taste aversion in the first experiment. Male Wistar rats received tetrodotoxin injections (10 ng) after drinking a coffee solution (0.5%), either in the lateral vestibular nucleus (LVN), the parabrachial nucleus or the dopaminergic area A8. Two days later they drank a cider vinegar solution (3%) not followed by injections. In a later choice test, only the group receiving the injection in the lateral vestibular nucleus displayed a coffee aversion. In a second experiment the role of the peripheral vestibular symptoms induced by LVN inactivation on substituting the aversive stimulus was explored in the same behavioral task. Rats anesthetized (Pentobarbital, 25 mg/kg) before tetrodoxin LVN blockade, that did not show peripheral symptoms, did not develop learned aversions. The coffee preference ratios did not differ to those animals receiving only anesthesia or those that remained undisturbed. These results showed that the bilateral blockade of the vestibular nuclei may induce peripheral vestibular symptoms that that may substitute the aversive stimulus in taste aversion learning.
Animals↗
Dehydroepiandrosterone prevents lipid peroxidation and cell growth inhibition induced by high glucose concentration in cultured rat mesangial cells.
The oxidative stress induced by high glucose concentration contributes to tissue damage associated with diabetes, including renal injury. Dehydroepiandrosterone (DHEA), the major secretory product of the human adrenal gland, has been shown to possess a multi-targeted antioxidant activity which is also effective against lipid peroxidation induced by high glucose. In this study we evaluated the effect of DHEA on the growth impairment which high glucose concentration induces in cultured rat mesangial cells. Primary cultures of rat mesangial cells were grown for 10 days in media containing either normal (i.e. 5.6 mmol/l) or high (i.e. 30 mmol/l) concentrations of glucose, without or with DHEA at different concentrations. The impairment of cell growth induced by high glucose was reversed by 100 nmol/l and 500 nmol/l DHEA, which had no effect on mesangial cells cultured in media containing glucose at the normal physiological concentration (5.6 mmol/l). In high-glucose cultured mesangial cells, DHEA also attenuated the lipid peroxidation, as measured by thiobarbituric acid reactive substances (TBARS) generation and 4-hydroxynonenal (HNE) concentration, and preserved the cellular content of reduced glutathione as well as the membrane Na+/K+ ATPase activity. The data further support the protective effect of DHEA against oxidative damage induced by high glucose concentrations, and bring into focus its possible effectiveness in preventing chronic complications of diabetes.
Aldehydes↗
Attitudes and practices of physicians and naturopaths toward herbal products, including use during pregnancy and lactation.
BACKGROUND: The popularity of complementary therapies continues to grow, and physicians are asked increasingly by their patients for information regarding these modalities. PURPOSE: To assess the impact of these trends on physicians and medical students, and compare their attitudes and practices with those of the naturopaths and their students, with particular interest in the use of herbal products during pregnancy and breastfeeding. MATERIALS AND METHODS: A detailed questionnaire was distributed by a medical student and a naturopathic student to a randomly selected group of physicians, medical students, naturopaths and naturopathic students. They were asked a variety of questions about their background, attitudes and practices concerning herbal products. RESULTS: Thirty-eight per cent of the questionnaires were returned, with a total of 242 respondents. Fifty-four per cent of physicians discussed complementary therapies with their patients, whereas 100% of naturopaths discussed conventional medicines with their patients. The most popular product recommended by both medical doctors and naturopaths was echinacea, followed by St John's Wort. Eighty-six per cent of physicians, 74% of medical students, 66% of naturopaths and 50% of naturopathic students think that complementary medical education should be incorporated into the standard medical curriculum. Only one physician actually recommended a herbal product to a pregnant patient compared with 49% of the naturopaths who felt comfortable doing so. CONCLUSIONS: Complementary medicine has become a reality, and physicians are recommending herbal products to their patients, although on a smaller scale than are naturopaths. However, the two most popular herbal products are the same in each group. Physicians are less likely to recommend herbal products to pregnant and breastfeeding women than are naturopaths.
Attitude of Health Personnel↗
Longitudinal change in the treatment of nausea and vomiting of pregnancy in Ontario.
BACKGROUND: Health problems associated with untreated nausea and vomiting of pregnancy (NVP) include maternal weight loss, dehydration, and electrolyte and acid-base disturbances. Negative social consequences include the deterioration of domestic, social and occupational function of the affected women. In 1994 it was documented that most physicians in Ontario tended not to treat women with NVP pharmacologically. PURPOSE: To investigate the longitudinal change in therapeutic practice of physicians, with respect to the treatment of NVP. SUBJECTS AND METHODS: A questionnaire was distributed to a randomly selected sample of physicians that included community-based family physicians, hospital-based family physicians, obstetricians and physicians who called the Motherisk Program for information. The participants were questioned about their choices in the pharmacological treatment of NVP. The data from the survey were compared with those from a previously published survey conducted in 1994. RESULTS: In 1999, 90.2% of physicians surveyed reported to have used pharmacological means to treat NVP. In 1999, 95% of the physicians surveyed reported to have prescribed doxylamine succinate-pyridoxine hydrochloric acid (Diclectin), and 11% prescribed dimenhydrinate (Gravol) to treat NVP. These results are significantly different from those found in 1994 (90% prescribed Gravol as the first choice). CONCLUSIONS: In 1999, temporally related to various educational efforts, physicians offered treatment for NVP more readily, including the drug recommended by the regulatory agency. These changes may explain in part the recently documented decrease in hospitalizations due to NVP in Canada.
Chi-Square Distribution↗
Functional blockade of the parabrachial area by tetrodotoxin disrupts the acquisition of conditioned taste aversion induced by motion-sickness in rats.
The role of the parabrachial area in conditioned taste aversion (CTA) induced by motion-sickness was studied in male Wistar rats. In the first experiment, one-trial conditioned taste aversion, to a 0.5% decaffeinated coffee solution, was induced by 30 min of vertical rotatory motion (80 rev./min) in intact rats. In the second experiment, reversible blockade of the neural activity in various brainstem sites was induced by bilateral intracerebral injections of tetrodotoxin (TTX) (10 ng/microl) after conditioning. Blockade of the parabrachial area, but neither A8 nor lateral vestibular nucleus, disrupted the acquisition of (CTA). The results are discussed in terms of an associative role of the parabrachial area in body rotation-induced taste aversion learning, as the area was intact during sensory processing and testing.
Analysis of Variance↗
Polyvariant mutant CFTR genes in patients with chronic pancreatitis.
Several authors have reported an association between mutations of the cystic fibrosis transmembrane conductance regulator gene (CFTR) and chronic pancreatitis. CFTR gene transcription and protein efficiency are influenced by two polymorphic loci, (TG)m and M470V, other than the T5 allele, whose role is already well-established. The TG11/T5 haplotype is commonly found in healthy subjects, while the TG12/T5/V470 and TG13/T5/V470 haplotypes are present in congenital bilateral absence of the vas deferens (CBAVD) patients. While the T5 allele is a mutation that is over-represented in patients with chronic pancreatitis, no data are available concerning the possible allelic preference at the other two polymorphic loci, (TG)m and M470V, in these patients. For this reason, we screened 39 patients with chronic pancreatitis for the most common CFTR mutations found so far in the Italian population; in addition, we examined the length of the polypyrimidine (poly-T) tract in intron 8, the (TG)m length and the M or V codon at position 470. CFTR mutations were found in 3 patients. Poly-T variant typing identified genotype T5/T7 in 5 patients and T5/T9 in 1 patient. Direct sequencing of intron 8 in patients with the T5 variant revealed the TG12/T5/V470//TG11/T7/V470 genotype in 5 patients and TG10/T9//TG11,T5 genotype in 1 patient. In patients with chronic pancreatitis, the T5 allele is frequently associated with TG12 and V470, a haplotype already reported in CBAVD cases and quite uncommon in healthy subjects.
Adult↗
Killer activity at different pHs against Cryptococcus neoformans var. neoformans serotype A by environmental yeast isolates.
Yeast isolates that share the same habitats as Cryptococcus neoformans var. neoformans serotype A in a restricted Mediterranean area were assayed in order to verify their killer activity against Cr. neoformans strains isolated from clinical and environmental sources. Many of the environmental yeast isolates expressed the killer phenomenon against the assayed strains of Cr. neoformans. Two species of Candida: Candida parapsilosis and Candida famata, and Pichia carsonii, were the most active killers at pH 4.6, 5.0 and 5.6 levels encountered in pigeon and canary guanos. Killer activity by C. parapsilosis is reported for the first time. The authors hypothesized that the killer phenomenon exerted by yeast species with heavy killer activity against Cr. neoformans would lend themselves for use as biological control agents against sensitive strains of Cr. neoformans when directly inoculated into the habitats of Cr. neoformans.
Animals↗