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Biomedical subjects

A Kang

Publications and source records attributed to A Kang.

16 recordsLinked to original sources

Complete genome sequence of the genetically tractable hydrogenotrophic methanogen Methanococcus maripaludis.

The genome sequence of the genetically tractable, mesophilic, hydrogenotrophic methanogen Methanococcus maripaludis contains 1,722 protein-coding genes in a single circular chromosome of 1,661,137 bp. Of the protein-coding genes (open reading frames [ORFs]), 44% were assigned a function, 48% were conserved but had unknown or uncertain functions, and 7.5% (129 ORFs) were unique to M. maripaludis. Of the unique ORFs, 27 were confirmed to encode proteins by the mass spectrometric identification of unique peptides. Genes for most known functions and pathways were identified. For example, a full complement of hydrogenases and methanogenesis enzymes was identified, including eight selenocysteine-containing proteins, with each being paralogous to a cysteine-containing counterpart. At least 59 proteins were predicted to contain iron-sulfur centers, including ferredoxins, polyferredoxins, and subunits of enzymes with various redox functions. Unusual features included the absence of a Cdc6 homolog, implying a variation in replication initiation, and the presence of a bacterial-like RNase HI as well as an RNase HII typical of the Archaea. The presence of alanine dehydrogenase and alanine racemase, which are uniquely present among the Archaea, explained the ability of the organism to use L- and D-alanine as nitrogen sources. Features that contrasted with the related organism Methanocaldococcus jannaschii included the absence of inteins, even though close homologs of most intein-containing proteins were encoded. Although two-thirds of the ORFs had their highest Blastp hits in Methanocaldococcus jannaschii, lateral gene transfer or gene loss has apparently resulted in genes, which are often clustered, with top Blastp hits in more distantly related groups.

Archaeal Proteins↗

Endothelial apoptosis as the primary lesion initiating intestinal radiation damage in mice.

Gastrointestinal (GI) tract damage by chemotherapy or radiation limits their efficacy in cancer treatment. Radiation has been postulated to target epithelial stem cells within the crypts of Lieberkühn to initiate the lethal GI syndrome. Here, we show in mouse models that microvascular endothelial apoptosis is the primary lesion leading to stem cell dysfunction. Radiation-induced crypt damage, organ failure, and death from the GI syndrome were prevented when endothelial apoptosis was inhibited pharmacologically by intravenous basic fibroblast growth factor (bFGF) or genetically by deletion of the acid sphingomyelinase gene. Endothelial, but not crypt, cells express FGF receptor transcripts, suggesting that the endothelial lesion occurs before crypt stem cell damage in the evolution of the GI syndrome. This study provides a basis for new approaches to prevent radiation damage to the bowel.

Animals↗

Quantitative trait loci affecting the behavior of A/J and CBA/J intercross mice in the elevated plus maze.

How allelic diversity affects neural mechanisms to produce behavioral variation is largely unknown. The elevated plus maze, consisting of open and closed arms, has been used as a model of behavioral variation in rodent exploration. Under dim illumination the nature of the sensory stimuli that influence arm choice is uncertain. Two inbred mouse strains, A/J (Tyrc/Tyrc, the albino phenotype, mutation in tyrosinase) with a strong preference for closed arm entry, and CBA/J (Pdebrdl/Pdebrdl, the retinal degeneration phenotype, mutation in the beta-subunit of rod cGMP phosphodiesterase), with a weak preference for open arm entry, were studied under varying light. Because behavioral differences persist under red light, variation in light perception is not likely to fully account for variation in arm choice. To identify genetic factors influencing arm choice (100 x Open arm entries/Total arm entries) quantitative trait loci analyses (QTL) were performed on (A/J x CBA/J)F2 mice. Two QTLs, one of which includes PDEB, were identified on Chr 5 (LOD > 10) and account for > 30% of the behavioral variation in arm preference. Tyr (Chr 7, 44 cM) was linked to closed arm entries but not arm preference, and is unlikely to be acting through a direct effect on light perception, because A/J arm entries were not affected by red light and there was no interaction with PDEB in the (A/J x CBA/J)F2 mice. Whether the candidate QTLs on Chr 5 affect arm choice through an effect on light perception is unknown, but phenotypic differences between F2 mice with retinal degeneration and CBA/J mice and F2 mice with albinism and A/J mice suggest that factors other than light sensitivity contribute to arm preference in these two strains.

3',5'-Cyclic-GMP Phosphodiesterases↗

Both maternal and fetal cell-free DNA in plasma fluctuate.

Elevations in the concentration of cell-free fetal DNA in maternal plasma have recently been determined in various pregnancy-related disorders, including preeclampsia, preterm labor, and polyhydramnios. In addition, almost 2-fold increments in cell-free fetal DNA levels have been recorded in pregnancies with certain aneuploid fetuses, in particular trisomy 21. These findings have led to the speculation that quantitative assessment of circulatory fetal DNA may be useful in the noninvasive prenatal diagnosis of certain fetal genetic constellations or pregnancy-related disorders. A premise for any quantitative analysis is that the quantity of the analyte being assayed does not vary greatly over time. As this aspect has not been examined for circulatory DNA levels, we examined these in normal healthy individuals as well as in pregnant women. Our data indicate that severalfold alterations in circulatory DNA amounts do occur over short periods of time. Of particular note is that we observed almost 2-fold variations in free fetal DNA levels over a period of 3 days, which are in a similar range to the elevations noted in aneuploid pregnancies. Our results, therefore, imply that caution should be used when using small increments in circulatory fetal DNA concentrations for potential diagnostic applications.

Cell-Free System↗

Fluctuation of maternal and fetal free extracellular circulatory DNA in maternal plasma.

OBJECTIVE: To examine whether concentrations of free extracellular fetal circulatory DNA in maternal plasma are stable or fluctuate. METHODS: Consecutive blood samples were drawn from 13 healthy nonpregnant volunteers and from 16 healthy pregnant women over 3 days. DNA was isolated from the plasma fraction and quantified by real-time polymerase chain reaction (PCR). RESULTS: In nonpregnant controls the total amount of cell free DNA fluctuated by an average of 13.5-fold. In samples obtained from pregnant women the amount of maternal cell free DNA varied by an average of 21.5-fold. Because ten of those women were pregnant with male fetuses, the concentration of free fetal DNA in these cases was determined by a real-time PCR assay for the Y chromosome. The mean variation in free fetal DNA levels in male fetuses was 2.2-fold. CONCLUSION: The degree of variation in free fetal DNA concentrations observed in this study was similar to published values, so these results imply that care should be exercised when considering quantitation of this fetal material for potential diagnostic or screening purposes.

Adult↗

[Fetal cells in maternal blood--their significance in non-invasive prenatal diagnosis and in etiologically determined diseases].

The first report of the occurrence of fetal cells in maternal circulation dates back to 1893 when the German pathologist Schmorl identified trophoblast cells in the lungs of women who had died from eclampsia. Till recently, however, the existence of fetal cells in maternal blood was a matter of considerable debate. The main reason for this dispute was the inability to successfully and reliably enrich for these cells, which has also hindered their clinical use. In the meantime, this issue has been addressed and the both fetal aneuploidies and single gene defects can now be detected in a non-invasive manner using fetal cells enriched from maternal blood.

Blood Cells↗

Lps(d)/Ran of endotoxin-resistant C3H/HeJ mice is defective in mediating lipopolysaccharide endotoxin responses.

C3H/HeJ inbred mice are defective in that they are highly resistant to endotoxic shock as compared with normal responder mice. Their B cells and macrophages do not respond significantly when exposed to lipopolysaccharide (LPS), whereas cells from the responder mice do. Using a functional assay, we previously isolated a cDNA, which encodes for Ran/TC4 GTPase. We now show that this gene is mutated in C3H/HeJ mice, which accounts for their resistance to endotoxin stimulation. Sequence analysis of independent mutant Lps(d)/Ran cDNAs isolated from splenic B cells of C3H/HeJ mice reveals a consistent single base substitution at position 870, where a thymidine is replaced with a cytidine. In situ hybridization maps the Lps(d)/Ran cDNA to mouse chromosome 4. By retroviral gene transfer, the wild-type Lps(n)/Ran cDNA but not the mutant Lps(d)/Ran cDNA can restore LPS responsiveness of C3H/HeJ cells. Adenoviral gene transfer in vivo with the mutant Lps(d)/Ran cDNA but not the wild-type Lps(n)/Ran cDNA rescues endotoxin-sensitive mice from septic shock. Thus Lps/Ran is an important target for LPS-mediated signal transduction, and the Lps(d)/Ran gene may be useful as a therapeutic sequence in gene therapy for endotoxemia and septic shock.

3' Untranslated Regions↗

Anatomic relation between the cervical pedicle and the adjacent neural structures.

STUDY DESIGN: An evaluation of the anatomic relation between the cervical pedicles and the adjacent neural structures. OBJECTIVES: To determine quantitatively the antomic relation of the cervical pedicles to the adjacent nerve roots and dural sac. SUMMARY OF BACKGROUND DATA: Transpedicular screw fixation in the cervical spine has been reported, but little quantitative data regarding the anatomic relation between the cervical pedicles and the surrounding neural structures are known. METHODS: Twenty cadavers were used for dissection to observe the relations of the cervical pedicles to the adjacent dural sac and nerve roots. After removal of whole posterior bony elements including the spinous processes, laminas, and lateral masses, the isthmus of the pedicles, the dural sac, and the nerve roots of C3-C7 were exposed. Direct measurements included the distance from the pedicle to the superior and inferior nerve roots and the dura. Also, the pedicle height and width were measured at its isthmus. RESULTS: No distance was found between the pedicle and the superior nerve root, nor between the pedicle and the dural sac at C3-C7 for all specimens. The mean distances between the pedicle and the inferior nerve roots for all specimens ranged from 1.4 to 1.6 mm. The mean pedicle heights and widths for all specimens at C3-C7 ranged from 6.0 to 6.5 mm and 4.7 to 5.3 mm, respectively. A significant difference between male and female specimens was noted in the pedicle heights for all levels measured (P = 0.001), and in the pedicle widths for the levels of C4 and C6 (P = 0.05). CONCLUSIONS: This study suggests that the incidence of neurologic injuries may be higher in screw penetration of the medial or superior cortex of the pedicle than in screw penetration of the inferior cortex of the pedicle.

Aged↗

Submaximal responses in calcium-triggered exocytosis are explained by differences in the calcium sensitivity of individual secretory vesicles.

A graded response to calcium is the defining feature of calcium-regulated exocytosis. That is, there exist calcium concentrations that elicit submaximal exocytotic responses in which only a fraction of the available population of secretory vesicles fuse. The role of calcium-dependent inactivation in defining the calcium sensitivity of sea urchin egg secretory vesicle exocytosis in vitro was examined. The cessation of fusion in the continued presence of calcium was not due to calcium-dependent inactivation. Rather, the calcium sensitivity of individual vesicles within a population of exocytotic vesicles is heterogeneous. Any specific calcium concentration above threshold triggered subpopulations of vesicles to fuse and the size of the subpopulations was dependent upon the magnitude of the calcium stimulus. The existence of multiple, stable subpopulations of vesicles is consistent with a fusion process that requires the action of an even greater number of calcium ions than the numbers suggested by models based on the assumption of a homogeneous vesicle population.

Animals↗

Catch-up growth in children treated with home enteral nutrition.

OBJECTIVE: This study was designed to determine the effect of home enteral nutrition on the outcomes of growth and the relationship between growth and entrance anthropometric criteria. METHODS: We reviewed the medical records of 78 consecutive children (median age, 20 months) who were enrolled in the home enteral feeding program at the Alberta Children's Hospital (Calgary, Alberta, Canada) between 1993 and 1995. Weights, heights, and weight-for-heights were expressed as Z scores, using the Centers for Disease Control and Prevention anthropometric growth curve software. To evaluate growth outcome, the total group was further subdivided using anthropometric criteria into appropriate, wasted, or stunted at the time of entry to the program. In a subgroup of 36 children on whom anthropometric data was available for a median length of 5.7 months, Z scores were compared at 3 points in time: before entry, at time of entry, and last follow-up. RESULTS: Patients were classified into five main groups: 11 (14%) had pulmonary disease, 26 (33%) had a gastrointestinal disorder, 21 (27%) had congenital defects, 10 (13%) had a neurologic disorder, and the remaining 10 (13%) had a variety of other illnesses, including malignancies and metabolic disorders. Patients were on the program for a median duration of 8.9 months. It was found that during the period of support within the program, enteral feeding was successful in improving weight-for-age Z scores by 0.42 standard deviations but the effect on height-for-age Z scores and weight-for-height Z scores did not reach significance for this population. The subgroup of 36 children on whom longitudinal anthropometric data was available before entering the program was found to have had a significant drop in weight Z scores between the time before program entry (median length of time, 5.7 months) and the time of program entry, which indicates that these children were falling off the growth curve before commencing enteral feeding. To evaluate growth outcome, the total group was further subdivided using anthropometric criteria into appropriate, wasted, or stunted at the time of entry to the program. In the group of appropriate growth patients, while in the program, 50% had catch-up growth for weight (positive change in Z scores) and 33% for height. In the wasted patients, 92% improved their weight percentile and 75% their height percentile. In the stunted group, 71% had catch-up growth for weight and 74% for height. CONCLUSION: We concluded that the enteral feeding program was able to promote catch-up growth or maintain growth along percentiles in the majority of children.

Adolescent↗

Cloning and expression of a single-chain antibody fragment specific for foot-and-mouth disease virus.

The gene for a single-chain antibody (VHK) to a conformational epitope on the type A12 foot-and-mouth disease virus (FMDV) particle was assembled and expressed in Escherichia coli. The VHK, purified from periplasmic extracts immunoprecipitated virus as efficiently as its parental monoclonal antibody (MAb) and exhibited the same binding specificity when tested against panel of natural and genetically engineered virus particles. The VHK neutralized type A12 virus in the presence of goat anti-mouse IgG; however, in the absence of the second antibody, only weak neutralizing activity was detected. Preliminary analysis of the mechanism of viral neutralization indicated that both the MAb and the VHK neutralize by the same mechanism. Small amounts of the VHK allowed infection of cells via Fc receptor-mediated adsorption in the presence of the second antibody. These data represent the first report of a single-chain neutralizing antibody for a picornavirus and provide insights into the mechanisms of viral neutralization and virus uptake.

Amino Acid Sequence↗

Propagation of an attenuated virus by design: engineering a novel receptor for a noninfectious foot-and-mouth disease virus.

To gain entry into cells, viruses utilize a variety of different cell-surface molecules. Foot-and-mouth disease virus (FMDV) binds to cell-surface integrin molecules via an arginine-glycine-aspartic acid (RGD) sequence in capsid protein VP1. Binding to this particular cell-surface molecule influences FMDV tropism, and virus/receptor interactions appear to be responsible, in part, for selection of antigenic variants. To study early events of virus-cell interaction, we engineered an alternative and novel receptor for FMDV. Specifically, we generated a new receptor by fusing a virus-binding, single-chain antibody (scAb) to intracellular adhesion molecule 1 (ICAM1). Cells that are normally not susceptible to FMDV infection became susceptible after being transfected with DNA encoding the scAb/ICAM1 protein. An escape mutant (B2PD.3), derived with the mAb used to generate the genetically engineered receptor, was restricted for growth on the scAb/ICAM1 cells, but a variant of B2PD.3 selected by propagation on scAb/ICAM1 cells grew well on these cells. This variant partially regained wild-type sequence in the epitope recognized by the mAb and also regained the ability to be neutralize by the mAb. Moreover, RGD-deleted virions that are noninfectious in animals and other cell types grew to high titers and were able to form plaques on scAb/ ICAM1 cells. These studies demonstrate the first production of a totally synthetic cell-surface receptor for a virus. This novel approach will be useful for studying virus reception and for the development of safer vaccines against viral pathogens of animals and humans.

Amino Acid Sequence↗

Effects of tetrandrine on lung-lipid contents of rats with experimental silicosis.

The wet weight, lipid contents and the percentages of PL components of lung tissues, SAM and AM of normal, Silicotic and TT treated rats were analysed. It was indicated that the wet weight, lipid contents of lung tissues, SAM and AM of Silicotic rats all were increased, and with the lapse of experimental period. In lung tissues the percentages of PC, PG and PI accounted for the total PL were increased and PS, S were decreased in comparison with those of normal rats. The PL percentages in SAM of Silicotic rats were not significantly different from those of normal rats. The wet weight, lipid contents of lung tissues, SAM and AM, percentages of PL components in lung tissues of TT treated rats were higher than that of normal rats and lower than that of silicotic rats. The percentages of PC, USPC accounted for the total PL in SAM of TT treated rats were lower while PI, PS,S were higher than that of normal and Silicotic rats. The other percentages of PL components were not significantly different from that of normal and silicotic rats.

Alkaloids↗

Celiac sprue and ulcerative colitis in three South Asian women.

The association of celiac disease and inflammatory bowel disease has not been described in females of South Asian descent. We report three women, aged 33, 43 and 46 years, of South Asian origin, with both ulcerative colitis and celiac disease.

Adult↗