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Biomedical subjects

K C Cheng

Publications and source records attributed to K C Cheng.

At least 37 records · Page 2Linked to original sources
PubMed1997

Patient-ventilator flow dyssynchrony: flow-limited versus pressure-limited breaths.

OBJECTIVES: Patient-ventilator flow dyssynchrony occurs when ventilator flow delivery is insufficient to meet patient demands. If sufficiently severe, flow dyssynchrony can produce significant imposed loads on ventilatory muscles. Flow dyssynchrony can be improved by increasing ventilator flow delivery. We hypothesized that the variable flow pressure-limited breath would be a better approach for matching patient flow demands than adjusting a set flow on a conventional volume-cycled breath. DESIGN: Clinical interventional study. SETTING: Medical intensive care unit. PATIENTS: Sixteen stable, mechanically ventilated patients receiving volume-cycled assist-control ventilation. INTERVENTIONS: Flow dyssynchrony was produced by reducing the set flow by 50%. Dyssynchrony was quantified by measuring the esophageal pressure time product during the assisted breath. Two strategies were then employed in an attempt to reduce the dyssynchrony. One strategy was to increase flow back to the initial set flow and then further increase flow by an additional 25% (VI strategy). The other strategy was to use a pressure-limited breath feature coupled to a volume assist breath (the P strategy). With the P strategy, the pressure limit was set at 75% and 100% of the static elastic recoil pressure at end-inspiration. MEASUREMENTS AND MAIN RESULTS: Pressure time product, intrinsic positive end-expiratory pressure, and the ventilatory pattern were measured with each strategy and were analyzed by analysis of variance. Induced baseline flow dyssynchrony, as measured by the pressure time product, was > 5 cm H2O/sec in ten of 16 patients. This dyssynchrony was significantly reduced by both the VI strategy and the P strategy, although the P strategy appeared to be more effective in those patients with the greatest baseline dyssynchrony. Baseline inspiratory time was also shortened by both the VI strategy and the P strategy; the VI strategy shortened baseline inspiratory time more than the P strategy. Baseline tidal volume, frequency, and intrinsic positive end-expiratory pressure were only minimally affected by either strategy. CONCLUSION: The pressure-limited, variable-flow approach to ventilator gas delivery appears to be more responsive to a vigorous patient effort than a fixed-flow approach.

Adult↗
PubMed1997

Applied PEEP during pressure support reduces the inspiratory threshold load of intrinsic PEEP.

OBJECTIVES: Mechanical ventilation in patients with obstructive airway disease (OAD) is associated with the development of dynamic hyperinflation and intrinsic positive end-expiratory pressure (PEEPi). One of the effects of this form of PEEPi is to act as an inspiratory threshold load that can produce ineffective breath triggering, dyspnea, and muscle fatigue. Recently it has been shown that applying PEEP in the ventilator circuit can reduce this imposed triggering load. We wished to investigate this further by studying patients with OAD being weaned with pressure support (PS) ventilation. Our first objective was to determine the prevalence and magnitude of this form of PEEPi in OAD patients who were clinically judged to be capable of triggering mechanical ventilatory breaths. Our second objective was to attempt to reduce the triggering load by applying circuit PEEP and then observe the response of patient-ventilator interactions during the patient-triggered, pressure-limited PS breath. DESIGN: Thirteen random patients with OAD who were receiving PS ventilation were studied by measuring airway pressures, airway gas flow, baseline esophageal pressure, esophageal pressure time products (PTP), and esophageal pressure changes before ventilator gas delivery began (delta Pes taken to represent PEEPi). Measurements were made at baseline and after stepwise increases in circuit PEEP up to the PEEPi. RESULTS: We found measurable PEEPi in all patients (average +/- SD of 9.54 +/- 4.3 cm H2O) and it was > 10 cm H2O in seven patients. As would be predicted, we observed progressive reductions in PEEPi as applied PEEP was given. We also observed that the component of patient effort (PTP) related to overcoming PEEPi also decreased, but the PTP related to tidal volume (VT) did not. The VT associated with the set PS thus did not change with application of PEEP, nor did the breathing frequency. CONCLUSION: PEEPi is common in OAD patients receiving mechanical ventilatory support. The imposed triggering load from PEEPi can be offset to large extent by circuit PEEP approaching the baseline PEEPi. Although total patient effort substantially falls with applied PEEP, the patient effort that combine with PS to effect VT does not.

Female↗
PubMed1997

Genetic dissection of vertebrate processes in the zebrafish: a comparison of uniparental and two-generation screens.

Genetic dissection is used to identify important genes in biological processes and is accomplished through the generation and study of mutations in model organisms. In diploid organisms, recessive mutations must be rendered hemizygous or homozygous for the mutant phenotype to be detected. In zebrafish (Danio rerio), this can be accomplished in two ways: (i) crosses between siblings who are potential carriers (in two-generation screens) and (ii) the generation of uniparental progeny. Two-generation screens have been the most productive to date but require substantial resources. However, uniparental screens involving haploids and half-tetrads (products of meiosis I) have also been productive, and require more modest resources. In genetic screens, the fraction of an average genome that is heterozygous (heterozygosity index) is inversely proportional to the likelihood that separate recessive mutations will be homozygous at the same time. Heterozygosity indices for haploid, half-tetrad, and two-generation screens are 0, 66, and 87.5%, respectively. Family sizes required to minimize bias in half-tetrad screens are also calculated. We conclude that gynogenetic half-tetrad screens are genetically robust and technically accessible to the independent researcher. The increasingly powerful genetic and experimental tools available for work with zebrafish can be used to address a broad range of questions in vertebrate biology.

Animals↗
PubMed1996

Factors affecting fidelity of DNA synthesis during PCR amplification of d(C-A)n.d(G-T)n microsatellite repeats.

The susceptibility of microsatellite DNA sequences to insertions and deletions in vivo makes them useful for genetic mapping and for detecting genomic instability in tumors. An in vitro manifestation of this instability is the production of undesirable frameshift products during amplification of (dC-dA)n x (dG-dT)n microsatellites in the polymerase chain reaction (PCR). These products differ from the primary product by multiples of 2 nucleotides. We have tested the hypothesis that factors known to affect the fidelity of DNA synthesis may affect (dC-dA)n x (dG-dT)n frameshifting during the PCR. Neither modifications of pH, dNTP concentration, and Mg++ concentration using Amplitaq, nor the use of thermophilic DNA polymerases including UITma, Pfu, Vent and Deep Vent significantly decreased the production of frameshift products during amplification. However, 3'-->5' exonuclease activity in thermophilic DNA polymerases inhibited the accumulation of PCR products containing non-templated 3' terminal nucleotides. Most interestingly, extension temperatures of 37 degrees C during amplification using the thermolabile DNA polymerases Sequenase 1.0, Sequenase 2.0, and 3'-->5' exonuclease-deficient Klenow fragment greatly decreased the production of frameshift products. This method can improve the resolution of heterozygous or mutant (dC-dA)n x (dG-dT)n alleles differing in size by one or two repeat units.

Base Sequence↗
PubMed1996

Brain stem auditory-evoked potentials of mice with experimental allergic encephalomyelitis.

Experimental allergic encephalomyelitis (EAE) is an experimental model for multiple sclerosis. In order to study autoimmune retrocochlear hearing loss, we performed brain stem auditory-evoked potential (BAEP) studies on EAE mice. The EAE was induced in B10.PL and (PL/J x SJL)F1 mice. In the BAEP study, all of the peak and interpeak latencies were prolonged significantly in the diseased mice. Hearing thresholds were slightly elevated in the immunized mice during the acute phase. Inflammatory and phagocytic cell infiltration, demyelination, and V beta 8.1, 8.2 T-cell receptor-positive cells were observed in the cochlear nerves or their proximity by histologic study. It is suggested that immunologic reactions identified with EAE also occurred in the cochlear nerve, and that these reactions were responsible for the hearing problems. It appears that EAE is a useful model system for studying autoimmune insults on the neural portion of the auditory system.

Animals↗
PubMed1995

Microsatellite-centromere mapping in the zebrafish (Danio rerio).

Ten (CA)n microsatellite simple sequence repeat (SSR) markers, 1, 2, 12, 14, 16, 18, 20, 22, 26, and 29, were used to show high chiasma interference and to determine centromere-marker map distances in the zebrafish (Danio rerio). Of these, SSR 12 exhibited no recombinant tetratypes among 175 half-tetrad embryos, placing this marker within 1 cM of the centromere of Linkage Group XVII. Fractions of heterozygous half-tetrads for the remaining nine markers ranged from 0.64 to 0.89. Of these, six recombinant fractions were more than 0.67 (P < 0.05), indicating strong chiasma interference during female meiosis in the zebrafish. Consistent with previous mapping data, SSRs 2 and 20 of Linkage Group VI were tightly linked. Half-tetrad analysis will allow the mapping of the remaining centromeres and may be useful in the mapping of new genes and mutations in the zebrafish.

Animals↗
PubMed1995

Substrate specificity, gene structure, and tissue-specific distribution of multiple human 3 alpha-hydroxysteroid dehydrogenases.

We have expressed in Escherichia coli functionally active proteins encoded by two human cDNAs that were isolated previously by using rat 3 alpha-hydroxysteroid dehydrogenase cDNA as the probe. The expressed proteins catalyzed the interconversion between 5 alpha-dihydrotestosterone and 5 alpha-androstane-3 alpha,17 beta-diol. Therefore, we name these two enzymes type I and type II 3 alpha-hydroxysteroid dehydrogenases. The type I enzyme has a high affinity for dihydrotestosterone, whereas the type II enzyme has a low affinity for the substrate. The tissue-specific distribution of these two enzymes was determined by reverse transcription polymerase chain reaction using gene-specific oligonucleotide primers. The mRNA transcript of the type I enzyme was found only in the liver, whereas that of the type II enzyme appeared in the brain, kidney, liver, lung, placenta, and testis. The structure and sequence of the genes encoding these two 3 alpha-hydroxysteroid dehydrogenases were determined by analysis of genomic clones that were isolated from a lambda EMBL3 SP6/T7 library. The genes coding for the type I and type II enzymes were found to span approximately 20 and 16 kilobase pairs, respectively, and to consist of 9 exons of the same sizes and boundaries. The exons range in size from 77 to 223 base pairs (bp), whereas the introns range in size from 375 bp to approximately 6 kilobase pairs. The type I gene contains a TATA box that is located 27 bp upstream of multiple transcription start sites. In contrast, the type II gene contains two tandem AP2 sequences juxtaposed to a single transcription start site.

3-Hydroxysteroid Dehydrogenases↗
PubMed1995

Localization of multiple human dihydrodiol dehydrogenase (DDH1 and DDH2) and chlordecone reductase (CHDR) genes in chromosome 10 by the polymerase chain reaction and fluorescence in situ hybridization.

Multiple human dihydrodiol dehydrogenases and human chlordecone reductase belong to the aldoketo reductase superfamily. These two enzymes are involved in the metabolism of xenobiotics, such as polycyclic aromatic hydrocarbons and pesticides. Recently we have isolated three closely related genes encoding two dihydrodiol dehydrogenases (DDH1 and DDH2) and the chlordecone reductase (CHDR). Mapping of the location of the genes was performed using the polymerase chain reaction using gene-specific primers to amplify gene sequences in human/hamster hybrid DNA. All three genes were found to be located on chromosome 10. In situ hybridization using a lambda clone as the probe further confirmed regional localization at 10p14-p15.

Alcohol Oxidoreductases↗
PubMed1995

Distribution of 3 alpha-hydroxysteroid dehydrogenase in rat brain and molecular cloning of multiple cDNAs encoding structurally related proteins in humans.

3 alpha-Hydroxysteroid dehydrogenase in the brain is responsible for production of neuroactive tetrahydrosteroids that interact with the major inhibitory gamma-aminobutyric acid receptor complexes. Distribution of 3 alpha-hydroxysteroid dehydrogenase in different regions of the brain in rats was evaluated by activity assay and by Western immunoblotting using a monoclonal antibody against liver 3 alpha-hydroxysteroid dehydrogenase as the probe. The olfactory bulb was found to contain the highest level of 3 alpha-hydroxysteroid dehydrogenase activity, while moderate levels of the enzyme activity were found in other regions such as cerebellum, cerebral cortex, hypothalamus and pituitary. Some activity was found in the rest of the brain such as amygdala, brain stem, caudate putamen, cingulate cortex, hippocampus, midbrain, and thalamus. The protein levels of 3 alpha-hydroxysteroid dehydrogenase in different regions of the brain as detected by Western immunoblotting are comparable to those of the enzyme activity. We used the rat cDNA as the probe to screen a human liver lambda gt11 cDNA library. A total of four different cDNAs were identified and sequenced. One of the cDNAs is identical to that of the human chlordecone reductase cDNA except that our clone contains a much longer 5'-coding sequence than previously reported. The other three cDNAs display high degrees of sequence homology to those of both rat 3 alpha-hydroxysteroid dehydrogenase and human chlordecone reductase. We are currently investigating the functional relationship between the enzymes encoded by these human cDNAs and 3 alpha-hydroxysteroid dehydrogenase.

3-Hydroxysteroid Dehydrogenases↗
PubMed1995

Rapid deoxyribonucleic acid analysis by allele-specific polymerase chain reaction for detection of mutations in the steroid 21-hydroxylase gene.

Rapid DNA analysis based on allele-specific polymerase chain reaction (PCR) using mutation site-specific primers was developed to detect mutations in the CYP21 gene known to cause steroid 21-hydroxylase deficiency. In contrast to the previous method, in which PCR of genomic DNA was followed by dot blot analysis with radioactive probes and multiple rounds of stripping and reprobing for each of the 8 most common mutation sites, the results using this new method were immediately visualized after the PCR run by ethidium bromide-stained agarose gel electrophoresis. Using allele-specific PCR, mutation(s) were identified on 148 affected chromosomes out of 160 tested. Although mutation(s) were identified on only one chromosome of 11 of these patients, their parents showed a consistent pattern on DNA analysis. The only exception was that in one family, in which the parents each had a detectable mutation, a mutation was detected on only one allele of the patient. Most likely there is a mutation in the patient's other allele that could have arisen de novo or was inherited from the parent and was not evident in the transmitting parent's phenotype. When compared with the dot blot procedure, allele-specific PCR is more rapid, less labor-intensive, and avoids the use of radioactivity.

Alleles↗
PubMed1995

Prenatal treatment and diagnosis of congenital adrenal hyperplasia owing to steroid 21-hydroxylase deficiency.

Since 1986, prenatal diagnosis and treatment of congenital adrenal hyperplasia due to 21-hydroxylase deficiency (21-OHD) have been carried out in 239 pregnancies. In 145, diagnoses were made by amniocentesis, whereas 77 were diagnosed using chorionic villus sampling. A newly developed, rapid allele-specific polymerase chain reaction was used for DNA analysis in some cases. Of 239 pregnancies evaluated, 37 babies were affected with classical 21-OHD. Of these, 21 were females, 13 of whom were treated prenatally with dexamethasone. Dexamethasone administered at or before 10 weeks gestation (9 affected female fetuses) was effective in reducing virilization. Seven fetuses had affected female siblings (Prader stages 1-5); 3 of these were born with entirely normal female genitalia, whereas the other 4 were significantly less virilized (Prader stages 1-2) than their siblings. The remaining 2 newborns had male siblings; 1 was born with normal genitalia, and the other was Prader stage 1. No significant or enduring side-effects were noted in either the mothers or the fetuses, indicating that dexamethasone treatment is safe. Prenatally treated newborns did not differ in weight, length, or head circumference from untreated unaffected newborns. Based on our experience, proper prenatal diagnosis and treatment of 21-OHD is effective in significantly reducing or eliminating virilization in the affected female. This spares the newborn female the consequences of genital ambiguity, i.e. genital surgery, sex misassignment, and gender confusion.

Adrenal Hyperplasia, Congenital↗
PubMed1995

Steroid 21-hydroxylase deficiency: genotype may not predict phenotype.

Steroid 21-hydroxylase deficiency is the most frequent cause of congenital adrenal hyperplasia. We have determined the 21-hydroxylase genotype in 197 patients with congenital adrenal hyperplasia owing to 21-hydroxylase deficiency and assessed phenotypic characteristics based on 1) genital status with respect to virilization in females, 2) ACTH stimulation tests to evaluate the secretion of androgens and 17-hydroxyprogesterone, and 3) salt deprivation tests to precisely describe the phenotype with respect to aldosterone deficiency and salt wasting. After dividing our patients into 26 21-hydroxylase gene mutation-identical groups, we found that, in general, the patient's phenotype matched the severity of the genotype. However, in 13 of these groups, the genotype did not always predict the phenotype, even within families. This study, has demonstrated that the 10 most common mutations observed in the 21-hydroxylase gene result in phenotypes that are not always concordant with the genotype.

Adrenal Hyperplasia, Congenital↗
PubMed1995

Aeromonas hydrophila lung abscess in a previously healthy man.

We present a case of multilocular lung abscess caused by Aeromonas hydrophila, found in both sputum and bronchial washing. No aspiration or other significantly contributory medical conditions were found. We believe this to be the first report of lung abscess caused by this organism in a previously healthy individual.

Aeromonas↗
PubMed1994

Structure of a gene coding for human dihydrodiol dehydrogenase/bile acid-binding protein.

The structure and sequence of the gene (DD/BABP) encoding a human dihydrodiol dehydrogenase/bile acid-binding protein (DD/BABP) were determined by analysis of genomic clones. Several overlapping clones containing parts of the gene were isolated from a lambda EMBL3 SP6/T7 library and characterized by restriction mapping and DNA sequencing. The gene spans approx. 16kb and consists of nine exons. The sizes of the exons range from 77 to 167 bp, whereas the intron sizes range from 375 to 3430 bp. The transcription start point (tsp) is located at -56 bp of the first ATG codon, as determined by primer extension. Neither a TATA box nor a CAT box was found upstream from the tsp. Southern blot analysis of the human genomic DNA, using the cDNA as the probe, revealed several additional hybridizing DNA bands, suggesting the existence of multiple related genes.

Amino Acid Sequence↗
PubMed1994

Structure and tissue-specific expression of the aldo-keto reductase superfamily.

We previously identified multiple proteins structurally related to 3 alpha-hydroxysteroid dehydrogenase in rat liver, lung, kidney, and testis ((1991) Arch. Biochem. Biophys. 291, 258-262). We further used these monoclonal antibodies to screen several lambda gt11 cDNA libraries derived from male rat liver, lung, and kidney. Five additional unique cDNA clones were isolated and sequenced; the proteins encoded by these cDNAs were found to exhibit 37-62% amino acid sequence homology to rat liver 3 alpha-hydroxysteroid dehydrogenase. Because these encoded proteins belong to the aldo-keto reductase superfamily, we named these proteins RAKa to RAKf. RAK represents rat aldo-keto reductase, and RAKa is the previously described rat liver 3 alpha-HSD. Northern blot analysis and reverse transcription-polymerase chain reactions were performed to examine their expression in various tissues. Only RAKe, which resembles human aldehyde reductase, was ubiquitously expressed in liver, kidney, lung, and other tissues, while the remaining mRNAs were found to have a more tissue- and sex-specific distribution. Genomic blot analysis showed complex, yet distinctive, restriction band patterns when different cDNAs were used as probes, suggesting that these cDNA clones are products of different genes and more related gene(s) may exist.

3-Hydroxysteroid Dehydrogenases↗