[Correlational verification of drug-induced liver injury with HLA-B*35:01 allele due to Polygonum multiflorum].

In order to verify the correlation between Polygonum multiflorum-induced liver injury and HLA-B*35 : 01 alleles, six hospitalized patients diagnosed with Polygonum multiflorum-induced liver injury (PM-DILI) were selected, and their clinicopathological data were collected. Simultaneously, blood HLA-B* 35 : 01 allele detection was performed. Among the six PM-DILI cases, 4 were male, aged 38.83 ± 10.13 years old. The types of liver injury were hepatocellular injury types in all, and the severity of liver injury in five cases was grade 3. The histological presentations were acute hepatitis and acute cholestatic hepatitis. PM-DILI cases were all HLA-B*35:01 carriers, with a carrier rate of 100%. This finding indicates that PM-DILI is significantly correlated with HLA-B*35:01 alleles. Therefore, HLA-B*35 : 01 alleles can be used as an important predictive indicator for PM-DILI.

[Research progress in the application of implantoplasty in the treatment of peri-implantitis].

Peri-implantitis is a kind of serious complication after tooth implantation. The absorption of alveolar bone lead to the exposure of rough implant surface, which would result in poor long-term therapeutic effect. Implantoplasty promises a better long-term therapeutic effect than bone augmentation technique. This article will introduce implantoplasty from two aspects: therapeutic effect and its influencing factor, safety and effectiveness.

Anesthetic practice for Caesarean section and factors influencing anesthesiologists' choice of anesthesia: a population-based study.

BACKGROUND: We examined the change in anesthetic practice for Caesarean section (CS) during the past decade and determined factors influencing anethesiologists' decisions. METHODS: The cases were identified from data retrieved from Longitudinal Health Insurance Database released by the Taiwan National Health Research Institute in 2008. Trend analysis was performed using logistic regression models. The decision tree analysis was performed using the chi-squared automatic interaction detector method and multivariable logistic regression analysis was performed to identify predictors of general anesthesia. RESULTS: A total of 25,606 women undergoing CS were studied. Logistic regression analyses revealed an upward trend of spinal anesthesia from 2000 to 2008 [57.8-67.5%, adjusted odds ratio (OR) = 1.06, 95% confidence interval (CI) = 1.05-1.07, P < 0.001] and a decreasing trend across time for both general and epidural anesthesia (5.5-3.9% and 36.7-28.6%; both OR < 1, both P < 0.001). Patterns of change in anesthetic practice across time for emergency and non-emergency CS were similar (all P < 0.05). Our data further demonstrated that early or threatened labor, a history of preeclampsia, antepartum hemorrhage, emergency CS, and previous CS were important predictors that influenced the anesthesiologists' choice of general anesthesia versus neuraxial anesthesia for women undergoing CS. CONCLUSIONS: Spinal anesthesia was the most common mode of anesthesia for CS deliveries in Taiwan during the past decade. Early or threatened labor, antepartum hemorrhage, emergency CS, previous CS, and preeclampsia are significant determinants of general anesthesia in CS deliveries.

Midazolam vs ondansetron for preventing postoperative nausea and vomiting: a randomised controlled trial.

We compared the prophylactic anti-emetic efficacy of midazolam and ondansetron in 90 patients scheduled for minor gynaecological (hysteroscopy) or urological (ureteroscopy) procedures planned to last 1-2 h under sevoflurane anaesthesia with spontaneous ventilation of the lungs via a laryngeal mask airway. Midazolam 2 mg or ondansetron 4 mg were administered intravenously 30 min before the end of surgery. The proportions of patients who experienced postoperative nausea and vomiting in the first 24 h (30% and 27% for the midazolam and ondansetron groups, respectively) were similar in the two groups. The incidence of postoperative nausea and vomiting was significantly smaller in both groups than predicted according to the patients' underlying risks (midazolam group: p = 0.018; ondansetron group: p = 0.017). There were no significant differences in average sedation scores or pain scores. Treatment using ondansetron for anti-emetic prophylaxis did not provide a superior benefit compared to midazolam in the present study.

Dynamic CT perfusion imaging with acetazolamide challenge for evaluation of patients with unilateral cerebrovascular steno-occlusive disease.

BACKGROUND AND PURPOSE: Perfusion CT (PCT) has the ability to measure quantitative values and produce maps of cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT). We assessed cerebral hemodynamics by using these parameters and acetazolamide challenge in patients with cerebrovascular steno-occlusive disease. METHODS: Fifteen patients underwent PCT with acetazolamide challenge. Comparison of mean CBF, CBV, and MTT was determined between hemispheres and before and after acetazolamide challenge. Hemispheric ratio and percent change due to acetazolamide administration were also calculated. Absolute values and percent changes 2 SDs outside the mean from the nonstenotic hemispheres were defined as abnormal. RESULTS: Significant decreases in CBF (-25.1%, P = .003) and significant increases in MTT (47.1%, P < .001) were found in stenotic hemispheres. After acetazolamide challenge, significant changes in CBF (-39.5%, P < .001) and MTT (92.9%, P < .001) were also seen. The acetazolamide test significantly decreased CBF hemispheric ratio (-20.3%, P < .001) and increased MTT hemispheric ratio (30.8%, P = .002), making both maps more asymmetric. Significance in CBF and MTT percent changes (P < .001 and P = .005, respectively) was found between hemispheres. When CBF percent changes were assumed to represent the true determinant of hemodynamic impairment, normal ranges of baseline MTT value and MTT percent changes demonstrated sensitivities of 66.7% and 100% and specificities of 58.3% and 75%, respectively, for detecting patients with hemodynamic impairment. CONCLUSION: Parameters obtained from PCT with acetazolamide are promising for the evaluation of cerebral hemodynamics in patients with cerebrovascular steno-occlusive disease.

The use of the GlideScope for tracheal intubation in patients with ankylosing spondylitis.

BACKGROUND: The GlideScopeVideo Laryngoscope is a new intubating device. The aim of the study was to investigate the use of the GlideScopefor tracheal intubation in patients with ankylosing spondylitis (AS) undergoing general anaesthesia. METHODS: Twenty AS patients were chosen to undergo tracheal intubation by the GlideScope. Preoperative airway assessments were carried out to predict the difficulty of tracheal intubation. Before intubation all patients were given a modified Cormack and Lehane (MCLS) grade and percentage of glottic opening (POGO) score by a separate anaesthetist using a Macintosh size 3 blade. The patients were then intubated, using the GlideScope, by a different anaesthetist during which the larynx was inspected and given another MCLS grade and POGO score. RESULTS: Twelve of the AS patients were judged to have had difficult intubation by preoperative airway assessment. Eleven of the twelve patients had MCLS grades III or IV by direct laryngoscopy and were considered to have had a difficult laryngoscopy. Naso-tracheal intubations by the GlideScope were successful on 17/20 occasions, including 8 of the 11 difficult laryngoscopy. The GlideScope improved the MCLS grade and POGO score in the majority of AS patients compared with direct laryngoscopy (P<0.01). CONCLUSIONS: The GlideScope provides a better laryngoscopic view than that of direct laryngoscopy. Most of the AS patients presenting with MCLS grade III or IV by direct laryngoscopy can be intubated successfully by the GlideScope. In elective patients with AS, awake fibreoptic intubation offers a higher level of security because it can be applied while maintaining spontaneous breathing. The use of GlideScope for tracheal intubation may be an alternative option in these patients who prefer their airway management under anaesthesia.

Molecular and structural analysis of immunoglobulin E-binding epitopes of Pen ch 13, an alkaline serine protease major allergen from Penicillium chrysogenum.

BACKGROUND: Through proteomic and genomic approaches we have previously identified and characterized an alkaline serine protease that is a major allergen (88% frequency of IgE binding) of Penicillium chrysogenum (Pen ch 13). OBJECTIVE: The aim of the present study is to identify the linear IgE-binding epitopes of Pen ch 13. METHODS: IgE-binding regions were identified by dot-blot immunoassay using 11 phage-displayed peptide fragments spanning the whole molecule of Pen ch 13. The minimal epitope requirements for IgE binding were further defined with overlapping peptides synthesized on derivatized cellulose membranes using SPOTs technology. The critical residues on the immunodominant epitopes were mapped through site-directed mutagenesis. The locations of the IgE epitopes identified were correlated with a three-dimensional structure of Pen ch 13. RESULTS: IgE antibodies in 35 serum samples reacted with at least one of the 11 peptide fragments of Pen ch 13. Peptide f-2n (residues 31-61) showed a high-intensity and the highest frequency (77%) of IgE binding. The frequencies of IgE binding to peptide f-4 (residues 93-133), f-1 (residues 1-37) and f-7 (residues 168-206) were 51%, 34% and 31%, respectively. SPOTs assay narrowed down the region of IgE binding of f-2n to residues 48-55 (GHADFGGR). Three, two and one epitope(s) that are four to nine amino acids in length, within f-4, f-1 and f-7, respectively, were found. Site-directed mutagenesis of Pen ch 13 revealed that substitution of His49 and/or Phe52 on Pen ch 13 with methionine resulted in proteins with drastic loss of IgE binding in seven sera tested. Proteins with amino acid replacements at residues 15-18 (RISS), or at residues 112 (I) and 116 (D) have lower IgE-binding reactivity in one of the two patient's sera tested. Substituting residues 117 (W), 119 (V) and 120 (K) also block most of the IgE binding in one of the two patient's sera tested. In addition, replacing residues 203 (V) and 204 (D) along with a deletion at residue 206 (Y) diminished the IgE binding in two serum samples tested. A model was constructed based on the structure of P. cyclopium subtilisin protease that has >90% (256 out of 283 amino acids) sequence identity with Pen ch 13. The major epitope (GHADFGGR) on Pen ch 13 formed a loop-like structure and was located at the surface of the allergen. CONCLUSIONS: Several linear IgE-reactive epitopes and their critical core amino acid residues were identified for the Pen ch 13 allergen. The major linear IgE-binding epitope, 48GHADFGGR55, formed a loop-like structure at the surface of the allergen. Substitution of His49 and/or Phe52 with methionine significantly reduced IgE-binding to Pen ch 13. Mapping of these results on a 3D model of the allergen provides valuable information about the molecular basis of allergenicity for Pen ch 13 and for designing specific immunotherapeutics.

Characterization of a novel Candida albicans 29 kDa IgE-binding protein--purification, cDNA isolation and heterologous expression of Cand a 3.

BACKGROUND: Candida albicans has been implicated in human allergic disorders. However, many of its immunoglobulin E (IgE)-reacting components have not yet been identified. The purpose of the present study is to characterize a novel 29 kDa IgE-binding protein from C. albicans. METHODS: The 29 kDa protein was partially purified and its tryptic digests subjected to mass spectrometric analysis. The cDNA encoding this protein was isolated and heterologously expressed in Escherichia coli. Monoclonal antibodies (MoAbs) were raised against the 29 kDa protein purified from C. abicans extracts. RESULTS: We isolated a 29 kDa IgE-reacting component from C. albicans. The protein was digested on-gel with trypsin and the masses of the resulting fragments were determined in a MALDI-TOF mass spectrometer. The data were searched against protein sequences deduced from the C. albicans genome. An open reading frame that possibly encodes the 29 kDa IgE-reacting component was identified. The cDNA corresponding to the open reading frame was isolated. It encodes a 236 residues protein that has 62% sequence identity to that of a hypothetical protein (YDR533c) from Saccharomyces cerevisiae. Conserved domain search suggests that the encoded protein belongs to the ThiJ/PfpI family. The cDNA isolated was inserted into a pQE-30 vector for protein expression in Escherichia coli. The recombinant protein can react with IgE antibodies in sera from asthmatic patients and two MoAbs that were generated against the purified native 29 kDa protein from C. albicans. CONCLUSIONS: We identified and cloned a novel 29 kDa IgE-reacting component (Cand a 3) from C. albicans. The recombinant proteins produced from this clone and the MoAbs prepared may be useful in the standardization of diagnostic extracts. They are also instrumental in elucidating the role of C. albicans in clinical allergy.

Molecular and immunological characterization of Pen ch 18, the vacuolar serine protease major allergen of Penicillium chrysogenum.

BACKGROUND: We have suggested previously that the 32 and 34 kDa major allergens of Penicillium chrysogenum (also known as P. notatum) are the vacuolar (Pen ch 18) and the alkaline (Pen ch 13) serine proteases, respectively, of P. chrysogenum. The purpose of this study is to characterize the 32 kDa allergen of P. chrysogenum and its immunoglobulin E (IgE)cross-reactivity with Pen ch 13 allergen. METHODS: The full-length cDNA of Pen ch 18 was isolated by reverse transcriptase-polymerase chain reaction and the 5'-rapid amplification cDNA end reaction. Recombinant Pen ch 18 was expressed as his-tagged proteins in Escherichia coli. Its reactivity with IgE and monoclonal antibodies against fungal serine protease allergens was analyzed by immunoblotting. The IgE cross-reactivity between Pen ch 18 and Pen ch 13 was analyzed by immunoblot inhibition. Overlapping recombinant fragments and synthetic peptides were used to map the B cell epitopes on Pen ch 18. RESULTS: In this study, we isolated a 1857 bp cDNA fragment containing an open reading frame of 494 amino acids that encodes the preproenzyme of Pen ch 18. Similar to other vacuolar serine proteases, this precursor appears to undergo N- and possibly C-terminal cleavage upon maturation. The his-tagged recombinant Pen ch 18 containing the putative sequence of the mature protein reacted with IgE antibodies in serum samples from asthmatic patients. In addition, IgE-binding to the 32 kDa major allergen of P. chrysogenum was inhibited when a positive serum sample was absorbed with recombinant Pen ch 18 before immunoblotting. Both inhibition and almost no inhibition of IgE-binding to the 32 kDa major allergen of Pen ch 18 were detected when eight positive serum samples were preabsorbed individually with purified Pen ch 13 before immunoblotting. The major IgE binding region was located in a fragment (PN1) encompassing the N-terminal 102 amino acid residues of the recombinant Pen ch 18. A dominant linear IgE epitope was further mapped within residues 73-95 (peptide PN1-e) of the N-terminally processed allergen. Monoclonal antibody FUM20 that reacts with Pen ch 18 but not with Pen ch 13 binds a synthetic peptide with sequence encompassing the N-terminal 23 residues of the recombinant Pen ch 18. Monoclonal antibody PCM39 that reacts with both Pen ch 13 and Pen ch 18 recognizes a peptide containing residues 132-154 of the allergen. CONCLUSIONS: Our results confirm that the Pen ch 18 allergen is a vacuolar serine protease of P. chrysogenum that matures through N- and possibly C-terminal processing. The finding that there are cross-reactive and allergen-specific IgE epitopes for Pen ch 18 and Pen ch 13 suggests that both major allergens should be included in clinically diagnostic P. chrysogenum extracts.

Novel mutation and prenatal sonographic findings of glutaric aciduria (type I) in two Taiwanese families.

Glutaric aciduria type I (GA I) is an autosomal recessively inherited inborn error with a defect of the enzyme glutaryl-CoA dehydrogenase (GCDH), which has never been diagnosed prenatally in Taiwanese patients. We present the prenatal sonographic findings and mutational analysis data of three children in two Taiwanese families. One patient from each family was diagnosed postnatally due to macrocephaly and neurological deterioration at 4 months and 10 months, respectively. The third child, sister of the first patient, was diagnosed prenatally at 11 weeks' gestation through chorionic villus sampling (CVS). Molecular analysis revealed that the fetus and child in Family 1 were homozygous for a common mutation, IVS10 -2A>C, which has not been reported in the Caucasian population. The patient in Family 2 was a compound heterozygote for IVS10 -2A>C and a novel mutation 749T>C (L238P). After genetic counseling, the couple decided to continue the second pregnancy. However, dilatation of quadrigeminal cistern (QC) and suspicious macrocephaly were noted at 30 weeks. Progressive dilatation of the QC associated with macrocephaly, fronto-temporal atrophy and wide space of perisylvian fissure were found in the follow-up scans. The affected girl was delivered at 37 weeks' gestation by cesarean section. Postnatal magnetic resonance imaging (MRI) studies confirmed the prenatal sonographic findings. With prenatal sonographic findings and mutational analysis presented in the present cases, the feasibility of prenatal diagnosis of GA I in high-risk pregnancy can not be overlooked.

Identification of a hepatic factor capable of supporting hepatitis C virus replication in a nonpermissive cell line.

Although hepatitis C virus E2 protein can bind to human cells by interacting with a putative viral receptor, CD81, the interaction alone is not sufficient to establish permissiveness for hepatitis C virus infection. Using an Epstein-Barr virus-based extrachromosomal replication system, we have screened through a human liver cDNA library and successfully identified a cDNA capable of supporting hepatitis C virus replication in an otherwise nonpermissive cell line. This cDNA encodes a protein exhibiting homology to a group of proteins derived from various evolutionarily distant species, including Oryza sativa submergence-induced protein 2A. The mRNAs encoding this factor are heterogeneous at the 5' ends and are ubiquitously expressed in multiple tissues, albeit in a very small amount. The longest mRNA contains an in-frame and upstream initiation codon and codes for a larger protein. This 5'-extended form of mRNA was detected in hepatocellular carcinoma, but not in normal liver tissue. Immunofluorescence analysis demonstrated that the hepatic factor was distributed evenly in cells, but occasionally formed aggregations in the peri- or intranuclear areas. In summary, we have identified a hepatic factor capable of supporting hepatitis C virus replication in an otherwise nonpermissive cell line. This factor belongs to a previously uncharacterized protein family. The physiological function of this protein awaits further study.

Low dose fentanyl and propofol improve the speed and quality of tidal-breathing induction techniques in sevoflurane anesthesia for adults.

BACKGROUND: The objective of this study was to investigate whether low dose fentanyl, with or without low dose propofol, as pretreatment agent/s is capable of speeding up and improving the quality of laryngeal mask airway (LMA) insertion in tidal-breathing induction technique with high-concentration sevoflurane. METHODS: One hundred and twenty patients were assigned to one of the three groups: Group S, induction with 8% sevoflurane only; Group F + S, 1.0 microgram/kg fentanyl prior to induction; and Group F + P + S, 1.0 microgram/kg fentanyl and 0.5 mg/kg propofol prior to induction. RESULTS: It was demonstrated that the time from administration of drug (drugs) to loss of eyelash reflex (P < 0.05, Group F + P + S vs. F + S; P < 0.01, Group F + P + S vs. S), to jaw relaxation (P < 0.05, Group F + P + S vs. S) and time taken for LMA insertion (P < 0.01, Group F + P + S vs. S) were all shorter in Group F + P + S, with fewer complications (coughing and involuntary movement) during induction, however, the first time success rate with LMA insertion did not significantly differ among the comparing groups. According to a postoperative inquiry (by questionnaire), there were significantly more patients in the Group F + P + S (57.5%) who considered the induction as pleasant (P < 0.05), of whom 75% expressed that they would be willing to undergo an induction of the same form again in the future (P < 0.05). This more positive rating may be related to the mild sedative effects of the agents given and shorter induction time, which significantly helped reduce the rate of recall of the unpleasant gas. CONCLUSIONS: The results of this study of LMA insertion, for ASA I or II adult patients undergoing the tidal-breathing technique with 8% sevoflurane, suggest that pretreatment with 1 microgram/kg fentanyl plus 0.5 mg/kg propofol is superior in comparison with either pretreatment with 1 microgram/kg fentanyl or absence of pretreatment.

cDNA cloning and immunologic characterization of Pen o 18, the vacuolar serine protease major allergen of Penicillium oxalicum.

Penicillium species are prevalent indoor airborne fungi that have been identified as causative agents of human extrinsic bronchial asthma. In the preparation of standardized diagnostic reagents, it is imperative to define the allergens of these ubiquitous fungi. Results from our previous study on P. oxalicum suggest that the 34-kd major immunoglobulin E-reacting component of this prevalent Penicillium species is probably a vacuolar serine protease. The purpose of the present study was to define this major P. oxalicum allergen (Pen o 18) through cDNA cloning and immunologic characterization. The cDNA of Pen o 18 was isolated through a combination of reverse transcriptase-polymerase chain reaction and 5'- and 3'-rapid amplification cDNA ends reactions. The primers used in these reactions were constructed according to the internal amino acid sequences of Pen o 18 and the conserved amino acid sequences of fungal serine proteases. Our results showed that a 1897-bp cDNA with an open reading frame of 503 residues was isolated for the proenzyme of Pen o 18. The encoded protein has a 16-residue signal peptide and a 119-residue prosequence. On maturation, the protein has an N-terminal glutamate that is the 136th residue encoded by the cDNA. Apparently the precursor also undergoes C-terminal processing with the cleavage of about 47 amino acids. The cDNA for Pen c 18 (the vacuolar serine protease allergen from P. citrinum ) was also isolated for comparison. Contrary to a previous report, the C-terminal region of Pen c 18 is similar to that of Pen o 18. Recombinant proteins (rPen o 18 and rPen c 18) with the putative mature N-termini and a his-tag were obtained by expressing the corresponding cDNAs in Escherichia coli. Serum samples from 7 asthmatic patients with immunoglobulin E reactivity to the 34-kd component of P. oxalicum also react to his-tagged recombinant Pen o 18. The presence of immunoglobulin E cross-reactivity between rPen o 18 and rPen c 18 was detected by immunoblot inhibition. Two monoclonal antibodies (PCM39 and FUM20) against fungal serine proteases react with rPen o 18, rPen c 18, and the 35/34-kd components in the corresponding crude fungal extracts. These components also react with immunoglobulin E antibodies in serum samples from asthmatic patients. In conclusion, results obtained confirm that the 34-kd major allergen of P. oxalicum is a vacuolar serine protease. The cDNAs of Pen o 18 and Pen c 18 encode precursor molecules that appear to undergo both N-terminal and C-terminal processing. Constructs beginning with mature N-terminal can be expressed in E. coli to produce recombinant polypeptides that are reactive to monoclonal antibodies or immunoglobulin E antibodies in serum samples from asthmatic patients. Results obtained may provide useful information and materials for preparation of standardized diagnostic reagents in clinical mold allergy.