Understanding the genetic relationships between Indonesian bambara groundnut landraces and investigating their origins.

A total of 170 bambara groundnut (Vigna subterranea) accessions were evaluated using both simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers generated using genotyping-by-sequencing (GbS), of which 56 accessions were collected from West and East Java. Principal coordinate analysis (PCoA), population structure, and cluster analysis suggest that the East Java accessions could be a result of the introduction of selected West Java accessions. In addition, the current Indonesian accessions were likely introduced from Southern Africa, which would have produced a very marked founding effect such that these accessions present only a fraction of the genetic variability that exists within this species.

Ethyl acetate fraction of the root of Rubia cordifolia L. inhibits keratinocyte proliferation in vitro and promotes keratinocyte differentiation in vivo: potential application for psoriasis treatment.

Psoriasis is a skin disease associated with hyperproliferation and aberrant differentiation of keratinocytes. Our previous studies have identified the root of Rubia cordifolia L. as a potent antiproliferative and apoptogenic agent in cultured HaCaT cells (IC(50) 1.4 microg/ml). In the present study, ethanolic extract of Radix Rubiae was fractioned sequentially with hexane, ethyl acetate (EA), n-butanol and water. EA fraction was found to possess most potent antiproliferative action on HaCaT cells (IC(50) 0.9 microg/ml). Mechanistic study revealed that EA fraction induced apoptosis on HaCaT cells, as it was capable of inducing apoptotic morphological changes. Annexin V-PI staining assay also demonstrated that EA fraction significantly augmented HaCaT apoptosis. In addition, EA fraction decreased mitochondrial membrane potential in a concentration- and time-dependent manner. The standardized EA fraction was formulated into topical gel and its keratinocyte-modulating action was tested on mouse tail model. EA fraction dose-dependently increased the number and thickness of granular layer and epidermal thickness on mouse tail skin, indicative of the keratinocyte differentiation-inducing activity. Taking the in vitro and in vivo findings together, the present preclinical study confirms that EA fraction is a promising antipsoriatic agent warranting further development for psoriasis treatment.

A binding protein for fatty acids in cytosol of intestinal mucosa, liver, myocardium, and other tissues.

A protein of molecular weight approximately 12,000 which binds long-chain fatty acids and certain other lipids has been identified in cytosol of intestinal mucosa, liver, myocardium, adipose tissue, and kidney. Binding is noncovalent and is greater for unsaturated than for saturated and medium-chain fatty acids. This protein appears to be identical with the smaller of two previously described cytoplasmic anion-binding proteins. Binding of long-chain fatty acids by this protein is greater than that of other anions tested, including sulfobromophthalein, and does not depend on negative charge alone. The presence of this binding protein may explain previously observed differences in intestinal absorption among fatty acids, and the protein may participate in the utilization of long-chain fatty acids by many mammalian tissues.

Medialization thyroplasty for cancer-related unilateral vocal fold paralysis.

OBJECTIVE: To justify the application of medialization thyroplasty in Chinese patients with symptomatic cancer-related unilateral vocal fold paralysis (UVFP). STUDY DESIGN AND SETTING: Retrospective chart review from February 2000 to March 2006. RESULTS: Eighty-seven Chinese patients undergoing medialization thyroplasty for UVFP were included; there were no significant differences between the cancer-related and benign groups in terms of the speech and swallowing rehabilitation outcome and the perioperative complication rate (P > 0.05). The median survival time of cancer-related UVFP patients from the date of medialization to death was 129 days. Age more than 65 years was identified as the only factor for a shorter survival period after medialization (P = 0.040). CONCLUSION: Medialization thyroplasty restores satisfactory speech and swallowing and has a low perioperative complication rate in Chinese patients with cancer-related UVFP. Postmedialization survival period was also reasonable. SIGNIFICANCE: Medialization thyroplasty is a justifiable treatment option for cancer-related UVFP.

The skin prick test results of 977 patients suffering from chronic rhinitis in Hong Kong.

OBJECTIVES: To evaluate the prevalence of aetiological allergens identifiable by skin prick test alone in patients suffering from chronic rhinitis in Hong Kong, and also compare the clinical history and symptoms of skin prick test-positive versus skin prick test-negative patients. DESIGN: Prospective study. SETTING: Otorhinolaryngology clinic in Queen Mary Hospital of Hong Kong. PATIENTS: A total of 977 patients suffering from chronic rhinitis were recruited into the study. Skin prick test was performed with a panel of allergens including house dust mites, cockroach, cat, dog, moulds, and pollens. MAIN OUTCOME MEASURES: Skin prick test results and their correlation with symptoms. RESULTS: Of the 977 patients, 651 (67%) had positive skin prick test reactions. The commonest allergen was house dust mite which was positive in 63% of the 977 patients and 95% of those 651 skin prick test-positive patients. The other allergens were in order of cockroach (23%), cat (14%), dog (5%), pollen (4%), and mould (3%). Compared with skin prick test-negative patients, skin prick test-positive patients were more likely to have earlier age of onset of the chronic rhinitis, association with asthma, more severe symptom in the morning, more severe symptoms of itchy nose, sneezing, nasal discharge, itchy eye, and watery eye. CONCLUSIONS: Identifiable aeroallergens could be detected in 67% chronic rhinitis patients by skin prick test alone. House dust mites were the most prevalent causative allergen. There were significant differences of patterns of clinical history and symptoms severity between skin prick test-positive and skin prick test-negative patients.

A multiobjective memetic algorithm based on particle swarm optimization.

In this paper, a new memetic algorithm (MA) for multiobjective (MO) optimization is proposed, which combines the global search ability of particle swarm optimization with a synchronous local search heuristic for directed local fine-tuning. A new particle updating strategy is proposed based upon the concept of fuzzy global-best to deal with the problem of premature convergence and diversity maintenance within the swarm. The proposed features are examined to show their individual and combined effects in MO optimization. The comparative study shows the effectiveness of the proposed MA, which produces solution sets that are highly competitive in terms of convergence, diversity, and distribution.

Cross-cultural adaptation and validation of the Chinese Voice Handicap Index-10.

OBJECTIVES/HYPOTHESIS: Recent developments in voice assessment propose the use of quality of life measurements. The Voice Handicap Index (VHI) is one of the most psychometrically robust and well-studied instruments among the various instruments for measuring quality of life. Two versions of VHI (VHI-30 and VHI-10) have been shown to be valid instruments for distinguishing dysphonic from nondysphonic individuals and also for documenting treatment effect for dysphonic patients. The VHI has been used worldwide; however, the psychometric properties of the Chinese version of VHI remains untested. This study aimed to investigate such properties of the Chinese VHI-30 and VHI-10 in the Hong Kong Chinese population. STUDY DESIGN: Psychometric analysis of the Chinese VHI-30 and VHI-10 in dysphonic patients and control subjects. METHODS: The original VHI-30 was translated into Chinese and was completed by 131 dysphonic patients and 54 nondysphonic individuals. The dysphonic patients also self-rated their dysphonic severity. RESULTS: Results showed high test-retest reliability and high item-total correlation for both Chinese VHI-30 and VHI-10. Both Chinese versions could be used to distinguish different dysphonic groups and between dysphonic and nondysphonic groups. Significant correlation was found between the VHI scores and the patients' self-rated dysphonic severity. CONCLUSION: The present study supported the original three-factorial structures of the VHI-30 and the use of the VHI for the Chinese population. It is shown that the Chinese VHI-10 is a strong representation of VHI-30 and is recommended for use in clinics because of its validity and ease of use by patients.

Reduction of opioid binding in neuroblastoma x glioma cells grown in medium containing unsaturated fatty acids.

Neuroblastoma x glioma cells NG108-15 were cultured in lipid-free medium supplemented with fatty acids of various chain length and unsaturation. Binding of 3H-labelled [DAla2]-[DLeu5]-enkephalin by membranes of cells grown in saturation fatty acids of different chain length was not significantly different from that of the control On the other hand, a proportional decrease of binding capacity with no change in residual receptor affinity was noticed when cells were cultured in medium containing fatty acids of increasing unsaturation. This decrease was time dependent and reached a maximum at about 48 h. Binding of [3H]dihydromorphine and [3H]naloxone was similarly affected. In contrast, when membranes of cells grown in normal medium were preincubated up to 3 h with unsaturated fatty acid and tested for opioid binding, no significant reduction was observed. Examination of the fatty acid composition of phospholipid from cells grown in linolenate indicated that a significant alteration of the acyl composition has occurred. To evaluate the underlying cause of this type of inhibition, the effect of linolenic acid on cell growth and protein synthesis was examined. When cells were cultured in 100 microM of this fatty acid, both growth and protein synthesis were retarded by 28% and 19%, respectively. Since opiate receptors are proteineous in nature, a reduction of protein synthesis may partially account for the loss of opioid binding activity. On the other hand, an increase of membrane fluidity is known to affect a number of cellular functions, including ligand-receptor recognition. Whether this can offer a satisfactory explanation for our observations remains to be established.

Antigenic determination fragments of alpha-momorcharin.

Alpha momorcharin is a protein isolated from the bitter gourd. It has a number of biological activities including induction of abortion, inhibition of tumor growth and anti-HIV. All these activities may be related to the ribosome-inhibiting activity of the protein. Repeated use of alphaMMC can elicit an antigenic response which may neutralize its biological activity. To overcome this problem, we need to know which part of the molecule is the antigenic determinant. In this study, we constructed a random fragment expression library from the alphaMMC cDNA and screened it with three anti-alphaMMC sera. A total of 9 positive clones were picked and sequenced. Based on the sequence information obtained, we were able to deduce three regions at which antibodies raised against native alphaMMC seem to interact. These regions are residues 1-14, residues 71-136 and residues 195-222. Mapping of these regions against a 3D model of alphaMMC indicates that they all are located on the surface of the molecule. As residues 71-136 are found to be in close proximity to the active site involved in ribosome inactivation, treatment with a monoclonal antibody directed to this area was shown to be effective in inactivating the inhibitory effect of alphaMMC on in vitro protein synthesis.

Expression of an immunologically reactive merozoite surface protein (MSP-1(42)) in E. coli.

The 42-kDa carboxyl-terminal processing fragment of Plasmodium falciparum merozoite surface protein-1 (PfMSP-1(42)) is one of the anti-malarial vaccine candidate antigens. In the present study, recombinant MSP-1(42) was expressed as a fusion protein in a novel E. coli host. The average yield of the recombinant protein was 48 mg/l of bacterial culture. The antigenicity and immunogenicity of the purified protein were evaluated by comparing the results with those obtained from a well-characterized recombinant MSP-1(42) (Bmp42) expressed in the baculovirus expression system previously described from our laboratory. We observed that there is a high degree of similarities between the two recombinant proteins. Based on the results from T and B cell response, in vitro parasite growth inhibition, as well as cross-reactivities with several well-characterized MSP-1 specific Mabs, the bacterial expressed protein is apparently comparable to Bmp42 in terms of immunoreactivities. Our results suggest that the bacterial expression system could be employed to express immunologically active recombinant MSP-1(42) at elevated levels. This system may be an attractive alternative for producing a protective vaccine for human use at lower cost.

Activation and inhibition of human alcohol dehydrogenase by monoclonal antibodies.

The human class I alcohol dehydrogenase isozyme beta 2 beta 2 was used as the antigen to raise monoclonal antibodies. Altogether seven lines of hybridomas secreting monoclonal antibodies were obtained. None of the antibodies was isozyme specific and all of them exhibited a similar affinity against all isozymes of the human class I ADH. Five out of the seven monoclonal antibodies had no effect on beta 2 beta 2 activity. Antibody G3 acted as a non-competitive inhibitor with a KI of 3 micrograms/ml at pH 7.5. Increasing pH was effective in reducing the level of inhibition. On the other hand, antibody 1D4 exhibited a pH-dependent activation of ADH activity. In the presence of this antibody, the pH optimum of beta 2 beta 2 was shifted from 9 to 8.5 and total activity was increased by 70% at this optimal pH. Kinetic analysis indicates that 1D4 probably acts as a non-competitive activator and may exert its action by interacting with the coenzyme binding site.

Cloning and sequencing of the grass carp (Ctenopharyngodon idellus) growth hormone gene.

An enriched lambda gt11 library for screening the grass carp (Ctenopharyngodon idellus) growth hormone gene was constructed using HindIII digested genomic DNA extracted from the spleen of the fish. Probing this library with a homologous cDNA, a clone carrying the growth hormone gene was obtained. The gene is 2501 bp long and consists of five exons and four introns. The sequence of nucleotide in the exons is almost identical to that of the cDNA except for ten positions. Analysis of the 5' sequence up to 1220 bp from the +1 position reveals the presence of a TATA box as well as a number of consensus regulatory sequences. Comparison of the grass carp growth hormone gene with those of other fish indicates that there is a high degree of homology with the gene from the common carp but not with those from the rainbow trout and Atlantic salmon.

Cloning of the cDNA of alpha-momorcharin: a ribosome inactivating protein.

Using a lambda gt11 cDNA library constructed from the seeds of the bitter melon (Momordica charantia), we have obtained a full length cDNA containing the entire sequence of alpha-momorcharin by immunoscreening. The length of this cDNA is 1044 basepairs long and it consists of an open reading frame coding for a polypeptide of 286 amino acids. The first 23 residues of this polypeptide probably code for a signal sequence. The N-terminal sequence of the deduced protein is exactly identical to that determined by peptide sequencing. The sequence identity between alpha-momorcharin and other ribosome inactivating proteins, such as trichosanthin and ricin A chain, is high, i.e., 34-63%. Examination of the predicted secondary structure of alpha-momorcharin and trichosanthin indicates that these proteins have regions of high structural similarities and this may account for the common biological activities that they share, viz., abortificant, immunosuppressive, antitumor and inhibition of HIV-1.

Expression of grass carp growth hormone by baculovirus in silkworm larvae.

A total of five recombinant Bombyx mori nuclear polyhedrosis viruses (BMNPV) carrying the grass carp (Ctenopharyngodon idellus) growth hormone (GH) cDNA were constructed in this study. Two of them were able to express the hormone up to a level of 12 microgram/ml medium when cultured B. mori cells were infected for 4 days. Inoculation of the viruses into silkworm (B. mori) host significantly increased the level of GH achievable. The amount of hormone produced per larva was estimated to be around 1 mg. The recombinant grass carp GH had immunological and biological activities similar to the native hormone. The N-terminal sequence of the recombinant hormone was the same as the native one, indicating that the fish signal peptide was correctly processed by the insect cells. Silkworm powder prepared from larvae infected with the recombinant virus was used as food supplement for fish. Compared with the control, this dietary supplement was effective in increasing the growth rate of juvenile carp.

Ethnic variation in sinonasal anatomy on CT-scanning.

Computed tomography (CT) scanning of the paranasal sinuses provides valuable information in assessing extent of disease and fine detailed anatomy prior to endoscopic sinus surgery. Awareness of the different anatomic variants of the bony sinonasal anatomy will help the rhinologic surgeon's orientation during the procedure. We conducted a study to look at the incidence of the anatomic variation within the lateral wall of the nose and sinuses and to determine if there is any ethnic difference in these variants between a cohort of 100 Caucasian patients undergoing endoscopic sinus surgery in London and 100 Chinese patients treated in Hong Kong. We compared the two groups with chi-square test and the significant areas are those with p value of less than 0.05. The results show a higher incidence of pneumatisation of the middle turbinate (concha bullosa) and paradoxical bending of the middle turbinate in the Caucasian population. The infraorbital and suprabullar cell development was greater in the Caucasian population though the incidence of sphenoethmoidal cells was much greater in the Chinese population. When asymmetry of the anterior ethmoidal roof was considered, the left was consistently the highest in both groups, though there was no difference in the depth of the cribiform niche between right and left or between Caucasian and Chinese. The incidence of bent uncinate process and of complete absence of a sinus was higher in the Chinese population. There was no difference in the presence of pneumatisation of the agger nasi, of the uncinate process, or of the anterior clinoid process. There was no difference in the presence of septation adjoining the carotid in the lateral wall of sphenoid or in the exposure of the optic nerve within sphenoid or posterior ethmoid sinuses. Although there is no evidence that variants of the sinonasal anatomy seen on CT Scan have a causative effect in the disease process; a knowledge of their presence is paramount in minimising the potential for surgical complications.

Cloning of trichosanthin cDNA and its expression in Escherichia coli.

Several cDNA clones coding for trichosanthin (TCS) have been isolated from a cDNA library prepared from the poly(A)+RNA of the root tuber of Trichosanthes kirilowii Maximowicz. The nucleotide sequence codes for a protein of 289 amino acids (aa) including a putative signal peptide of 23 aa and an extra 19 aa at the C terminus; the latter two have not been found in TCS obtained from the natural product [Collins et al., J. Biol. Chem. 265 (1990) 8665-8669]. Recombinant TCS (reTCS) was synthesized in Escherichia coli, in which the cDNA without the signal sequence was expressed under the control of the trc promoter; reTCS was detected by a rabbit anti-TCS antiserum.

Sensorineural hearing loss in patients treated for nasopharyngeal carcinoma: a prospective study of the effect of radiation and cisplatin treatment.

PURPOSE: The pattern of sensorineural hearing loss (SNHL) after primary treatment for nasopharyngeal carcinoma (NPC) was studied, and the effect of cisplatin, radiotherapy does, and fractionation were evaluated. METHODS AND MATERIALS: One hundred thirty-two patients, 227 ears, and 1100 audiogram reports were analyzed. Radiotherapy dose ranged from 59.5 to 76.5 Gy. Fifty-two patients received preirradiation cisplatin, total dose 100-185 mg/m(2). Serial postirradiation bone conduction thresholds at 0.5 kHz, 1 kHz, 2 kHz, and 4 kHz were compared with pretreatment thresholds at respective frequencies. Increase of at least 15 dB was considered as significant and was further grouped as transient or persistent SNHL. Univariate and multivariate analyses were performed to identify predicting factors for persistent SNHL. RESULTS: At median follow-up of 30 months, 24.2% of ears developed persistent SNHL. High frequency was more affected than low frequencies, 22 vs. 5.3%. Males were more affected than females, 29.4 vs. 15.5%, p = 0.0132. Incidence of persistent SNHL increased with age, with 0, 17.2, and 37.4% of patients aged under 30, between 30-50 and over 50 affected, respectively, p = 0.0001. High incidence was found in patient with postirradiation serous otitis media (SOM), 46.9%. Chemotherapy with cisplatin and radiation dose or fractionation had no significant effect. Multivariate analysis confirmed age, sex, and postirradiation SOM as significant prognostic factors for persistent SNHL. CONCLUSIONS: Transient and persistent SNHL occurred after radiotherapy, more commonly affecting high frequency. A low dose of preirradiation cisplatin did not increase the risk. A dose fractionation effect of radiotherapy was not confirmed in this study.

Analysis of Clonality in Kimura's Disease.

Kimura's disease is a chronic inflammatory disorder of unknown etiology. A 62-year-old man presented with asymptomatic cervical lymphadenopathy associated with eosinophilia and increased serum immunoglobulin E. Excision biopsy showed Kimura's disease. Three years later another groin lymph node appeared and showed similar pathologic features. Polymerase chain reaction for Ig heavy chain and T-cell receptor (TCR) genes on DNA extracted from the cervical lymph node showed smear patterns. However, polymerase chain reaction for TCRdelta gene showed a clonal rearrangement. Sequencing showed a complete VDJ rearrangement (Vdelta1-N-Ddelta2-N-Jdelta), confirming the presence of a clonal T cell population. The same clonal TCRdelta rearrangement was amplified by polymerase chain reaction from the groin lymph node biopsied 3 years later. These results showed that the primary and recurrent lesions were biologically related. Furthermore, the presence of identical T cell clones in different sites and at different times suggested that a clonal T cell population might have contributed to the pathogenesis in this case of Kimura's disease.

Opioid-like activity in the cerebrospinal fluid of pain patients treated by electroacupuncture.

Thirteen patients with pain from various causes were treated by electroacupuncture for 30 min. Cerebrospinal fluid (CSF) was obtained before and after treatment. Opioid-like substances in the CSF were fractionated by high pressure liquid chromatography and assayed by competitive receptor binding using a mu-specific radioligand, [D-ala2, MePhe4, gly-ol5]-enkephalin (DAGO). Opioid activity, associated with a fraction, eluted at 18-20% acetonitrile, consistently showed an increase in level after acupuncture. Two other fractions eluted at larger concentrations of acetonitrile also increased significantly after acupuncture; however the increase was not consistently observed in every patient. Measurements of beta-endorphin and dynorphin by radioimmunoassay indicated that 80 and 60% of the patients, respectively, had a higher level of these peptides after acupuncture. The nature of the opioid activity, eluted at 18-20% acetonitrile is unknown; however a small amount of it could be found in various parts of the brain of rat.

Effects of Ca(2+)-buffer concentration and stimulus interval on the voltage dependence and timecourse of calcium-release-dependent inward current in rabbit atrial myocytes.

To investigate the kinetics of the inward Na-Ca exchange tail current activated by internal calcium in rabbit atrial cells, the whole-cell patch-clamp technique with intracellular perfusion was used. We recorded the inward phase of this current during repolarizations following a brief 2-5 ms depolarizing pulse to +40 mV from a holding potential of -70 mV. Peak activation of the current occurs about 10 ms from the beginning of the depolarizing pulse, and it decays spontaneously with a slow timecourse. The voltage dependence of the process that activates the inward current from -40 mV to +40 mV has a very steep slope between -40 and -20 mV and then virtually saturates between -10 mV and +40 mV. The voltage dependence of the process that activates the inward current is steeper than that which activates the sarcolemmal calcium current, iCa.L, and the timecourse of the current relaxation is much slower at low-frequency stimulation and when using low concentrations of Ca-buffer. The magnitude and timecourse of the calcium transients estimated by the inward tail current are smaller and faster, and the slow component of decay was abolished by the presence of high intracellular concentrations of Ca-buffer or by high frequency stimulation. These observations suggest that calcium release from the sarcoplasmic reticulum may be triggered by only a small fraction of the sarcolemmal calcium current.