Fine mapping of a new common bean anthracnose resistance gene (Co-18) to the proximal end of Pv10 in Indian landrace KRC-5.

KEY MESSAGE: Mapping and fine mapping of bean anthracnose resistance genes is a continuous process. We report fine mapping of anthracnose resistance gene Co-18 which is the first anthracnose gene mapped to Pv10. The discovery of resistance gene is a major gain in the bean anthracnose pathosystem research. Among the Indian common bean landraces, KRC-5 exhibit high levels of resistance to the bean anthracnose pathogen Colletotrichum lindemuthianum. To precisely map the anthracnose resistance gene, we used a Recombinant Inbred Line (F2:9 RIL) population (KRC-5 × Jawala). The inheritance test revealed that KRC-5 carries a dominant resistance gene temporarily designated as Co-18. We discovered two RAPD markers linked to Co-18 among 287 RAPD markers. These RAPD markers were eventually developed into SCARs (Sc-OPR15 and Sc-OPF6) and flank Co-18 on chromosome Pv10 at a distance of 5.3 and 4.2 cM, respectively. At 4.0-4.1 Mb on Pv10, we detected a SNP (single-nucleotide polymorphism) signal. We synthesized 58 SSRs and 83 InDels from a pool of 135 SSRs and 1134 InDels, respectively. Five SSRs, four InDels, and two SCARs were used to generate the high-density linkage map, which led to the identification of two SSRs (SSR24 and SSR36) that are tightly linked to Co-18. These two SSRs flank the Co-18 to 178 kb genomic region with 13 candidate genes including five NLR (nucleotide-binding and leucine-rich repeat) genes. The closely linked markers SSR24 and SSR36 will be used in cloning and pyramiding of the Co-18 gene with other R genes to develop durable resistant bean varieties.

RT-PCR based detection of Pepper mild mottle virus from capsicum seeds and seed transmission assay.

Pepper mild mottle virus (PMMoV), a Tobamovirus from Virgaviridae family, is highly contagious and transmitted by seeds as well as soil in nature. PMMoV has become a greater threat to capsicum cultivation worldwide. To develop an indigenous, rapid, and sensitive protocol for routine detection of PMMoV from seeds, the sensitivity of DAS-ELISA and RT-PCR was compared in the present study. The infected seeds of California Wonder were included in the study. Through DAS-ELISA the virus was successfully detected from 20 mg of seeds. However, using RT-PCR, we were able to detect the virus even from one infected seed with reproducibility. In the present study, vertical seed transmission of the test virus was investigated by employing a grow-out test under greenhouse conditions as well as directly through RT-PCR omitting the grow-out test in three capsicum cultivars. Based on symptoms observations in grow out test, seed transmission was observed in the 3 capsicum cultivars viz., California Wonder (63.04%), Yolo Wonder (33.80%) and Doux des LAndes (33.30%). Through RT-PCR it was estimated to be 55.56% (California Wonder), 28.96% (Yolo Wonder), and 40.64% (Doux des Landes), respectively. Thus, indicating 100% seed-to-seedling PMMoV transmission and reliability of RT-PCR in direct PMMoV detection from seeds. Even a small percentage of infected seed has the potential to greatly increase the PMMoV inoculum in the field and result in 100% plant infection. Therefore, we suggest using the established procedure for PMMoV detection right from the seed. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-023-00807-0.

North-Western Himalayan Common Beans: Population Structure and Mapping of Quantitative Anthracnose Resistance Through Genome Wide Association Study.

Common bean (Phaseolus vulgaris L.) is an important legume crop of north-western (NW) Himalayan region and the major disease that causes catastrophic loss to the crop is anthracnose, which is caused by Colletotrichum lindemuthianum. The pathogen is highly diverse and most of the commercial cultivars are susceptible to different races prevalent in the region. The lack of information on the genomic regions associated with anthracnose resistance in NW Himalayan common bean population prompted us to dissect Quantitative Resistance Loci (QRLs) against major anthracnose races. In this study, 188 common bean landraces collected from NW region were screened against five important anthracnose races and 113 bean genotypes showed resistance to one or multiple races. Genotyping by sequencing (GBS) was performed on a panel of 192 bean lines (4 controls plus 188 Indian beans) and 22,589 SNPs were obtained that are evenly distributed. Population structure analysis of 192 bean genotypes categorized 188 Indian beans into two major clusters representing Andean and Mesoamerican gene pools with obvious admixtures. Many QRLs associated with anthracnose resistance to Indian C. lindemuthianum virulences (race 3, 87, and 503) are located at Pv04 within the gene models that encode typical resistance gene signatures. The QRLs associated with race 73 are located on Pv08 and overlaps with Co-4 anthracnose resistance gene. A SNP located at distal end of Pv11 in a gene model Phvul.011G202300 which encodes a LRR with a typical NB-ARC domain showed association with race 73 resistance. Common bean genomic regions located at Pv03, Pv09, and Pv11 showed association with resistance to anthracnose race 2047. The present study showed presence of many novel bean genomic regions associated with anthracnose resistance. The presence of Co-4 and Co-2 genes in our material is encouraging for breeding durable anthracnose resistant cultivars for the region.

Heterologous expression of pepper mild mottle virus coat protein encoding region and its application in immuno-diagnostics.

Pepper mild mottle virus (PMMoV), a tobamovirus of family Virgaviridae affects the quality and quantity of Capsicum. PMMoV is highly contagious, capable of transmitting through infected seeds and soil. Symptoms are more severe when crop is infected at young stage but remain unnoticed when infection takes place at maturity. Therefore, cost effective diagnostic techniques are required for timely and accurate detection of virus. In present study, coat protein encoding region of PMMoV-HP1 isolate was cloned into expression vector system, pET28a and expressed in BL21, a protease deficient strain of Escherichia coli. The PMMoV-HP1 pathotype was identified as PMMoV-P12 on the basis of coat protein amino acid sequence analysis in our previous study. The overexpression of recombinant coat protein of 26 kDa, corresponding to the expected 6X Histidine tag fused recombinant protein was purified using Ni-NTA columns from insoluble fraction. For antisera production, the purified recombinant protein was dialyzed ~ 24 h to remove urea and then used for raising polyclonal antisera. The specificity and sensitivity of antiserum obtained was demonstrated using different dilutions of antiserum for western blot assay and direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA). In Western blot assay, the test antiserum reacted strongly both with PMMoV-CP in purified protein and native CP in crude sap from PMMoV infected pepper plants, whereas no reaction was observed with healthy plant sap. In DAC-ELISA antiserum dilution up to 1:1000 was capable of detecting the virus in infected sample. The absence of any cross reactivity of test antiserum was confirmed against tobacco mosaic virus, cucumber mosaic virus, tomato spotted wilt virus, pepper veinal mottle virus, potato virus Y and tomato yellow leaf curl virus antigen, known to infect capsicum.

Status of F-18 fluorodeoxyglucose uptake in normal and hibernating myocardium after glucose and insulin loading.

OBJECTIVE: F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) has been increasingly used in myocardial viability imaging. In routine PET viability studies, oral glucose and intravenous insulin loading is commonly utilized. In an optimal study, glucose and insulin loading is expected to cause FDG uptake both in hibernating and normal myocardium. However, in routine studies it is not uncommon to see absent or reduced FDG uptake in normal myocardium. In this retrospective study we further analyzed our PET viability images to evaluate FDG uptake status in myocardium under the oral glucose and intravenous insulin loading protocol that we use in our hospital. METHODS: Patients who had both myocardial perfusion single photon emission computed tomography (SPECT) and FDG PET cardiac viability studies were selected for analysis. FDG uptake status in normal and abnormal myocardial segments on perfusion SPECT was evaluated. Based on SPECT and PET findings, patients were divided into two main groups and four subgroups. Group 1 included PET viable studies and Group 2 included PET-nonviable studies. Subgroups based on FDG uptake in normal myocardium were 1a and 2a (normal uptake) and 1b and 2b (absent or significantly reduced uptake). RESULTS: Seventy-one patients met the inclusion criteria. Forty-two patients were PET-viable and 29 were PET-nonviable. In 33 of 71 patients (46.4%) there was absent or significantly reduced FDG uptake in one or more normal myocardial segments, which was identified more in PET-viable than PET-nonviable patients (59.5% vs. 27.5%, p = 0.008). This finding was also more frequent in diabetic than nondiabetic patients (53% vs. 31.8%), but the difference was not significant (p = 0.160). CONCLUSIONS: In nearly half of our patients, one or more normal myocardial segments showed absent or significantly reduced FDG uptake. This finding, particularly if it is diffuse, could be from suboptimal study, inadequacy of current glucose and insulin loading protocols, or various other patient-related causes affecting FDG uptake both in the normal and hibernating myocardium. In cases with significantly reduced FDG uptake in normal myocardium, PET images should be interpreted cautiously to prevent false-negative results for viability.

Integrated Translatome and Proteome: Approach for Accurate Portraying of Widespread Multifunctional Aspects of Trichoderma.

Genome-wide studies of transcripts expression help in systematic monitoring of genes and allow targeting of candidate genes for future research. In contrast to relatively stable genomic data, the expression of genes is dynamic and regulated both at time and space level at different level in. The variation in the rate of translation is specific for each protein. Both the inherent nature of an mRNA molecule to be translated and the external environmental stimuli can affect the efficiency of the translation process. In biocontrol agents (BCAs), the molecular response at translational level may represents noise-like response of absolute transcript level and an adaptive response to physiological and pathological situations representing subset of mRNAs population actively translated in a cell. The molecular responses of biocontrol are complex and involve multistage regulation of number of genes. The use of high-throughput techniques has led to rapid increase in volume of transcriptomics data of Trichoderma. In general, almost half of the variations of transcriptome and protein level are due to translational control. Thus, studies are required to integrate raw information from different "omics" approaches for accurate depiction of translational response of BCAs in interaction with plants and plant pathogens. The studies on translational status of only active mRNAs bridging with proteome data will help in accurate characterization of only a subset of mRNAs actively engaged in translation. This review highlights the associated bottlenecks and use of state-of-the-art procedures in addressing the gap to accelerate future accomplishment of biocontrol mechanisms.

Molecular cloning and characterization of ech46 endochitinase from Trichoderma harzianum.

In the present study, endochitinase of T. harzianum isolate-ThHP3 induced against mycelium of F. oxysporum was cloned, sequenced and characterized. The complete nucleotide sequence contained an ORF of 1293bp corresponding to 430 amino acids with 46kDa molecular weight and theoretical pI 5.59. The precursor protein contained 22 amino acids long signal peptide at N terminus. The domain architecture of endochitinase showed low complexity regions, presence of 1W9P domain specific to cyclopentapeptide and lack of carbohydrate binding modules. The ligand binding site of ech46 endochitinase was constituted by 10 amino acids. The cDNA encoding ech46 endochitinase was ligated into pET28a vector and transformed to E. coli BL21. The predicted molecular weight of recombinant endochitinase without signal peptide was 49.4kDa with a theoretical pI 6.67. SDS-PAGE analysis of purified 6xHis tagged protein showed a single band of 49kDa. The refolded enzyme was active under acidic conditions with a temperature and pH optima of 50°C and 4. Km and Vmax for recombinant endochitinase using 4-pNP-(GlcNAc)3 were 315.2±0.36µM and 0.140±0.08µMmin-1, respectively and the calculated kcat was 6.44min-1. The RT-qPCR revealed induction of ech46 by phytopathogenic fungi.

Identification and molecular characterization of Bean yellow mosaic virus infecting French bean in Himachal Pradesh.

French bean (Phaseolus vulgaris L.), is one of the most widely grown vegetable crop. Disease samples showing yellow mosaic symptoms on leaves and pods were collected from Himachal Pradesh and inoculated on common bean cv. Jawala through sap inoculation. The virus successfully transmitted by mechanical inoculation produced yellow mosaic, leaf distortion, curling, wrinkling of leaves followed by stunting of plants. The identity of the virus as Bean yellow mosaic virus (BYMV) was established through Double antibody sandwich-enzyme linked immunosorbent assay, multiple sequence alignment and phylogenetic analysis of the coat protein gene sequence amplified by reverse transcription-polymerase chain reaction. The cp gene contained 819 nucleotides potentially coding for 273 amino acids. The sequence showed 83-99 % nucleotide and 89-99 % amino acid sequence identities with other BYMV isolates/strains and shared maximum identity with BYMV strain reported from Gladiolus sp. in Japan. This study constitutes the first report of BYMV occurrence on P. vulgaris in Himachal Pradesh.

Improved n-butanol production by a non-acetone producing Clostridium pasteurianum DSMZ 525 in mixed substrate fermentation.

The kinetics of growth, acid and solvent production in batch culture of Clostridium pasteurianum DSMZ 525 were examined in mixed or mono-substrate fermentations. In pH-uncontrolled batch cultures, the addition of butyric acid or glucose significantly enhanced n-butanol production and the ratio of butanol/1,3-propanediol. In pH-controlled batch culture at pH = 6, butyric acid addition had a negative effect on growth and did not lead to a higher n-butanol productivity. On the other hand, mixed substrate fermentation using glucose and glycerol enhanced the growth and acid production significantly. Glucose limitation in the mixed substrate fermentation led to the reduction or inhibition of the glycerol consumption by the growing bacteria. Therefore, for the optimal growth and n-butanol production by C. pasteurianum, a limitation of either substrate should be avoided. Under optimized batch conditions, n-butanol concentration and maximum productivity achieved were 21 g/L, and 0.96 g/L × h, respectively. In comparison, mixed substrate fermentation using biomass hydrolysate and glycerol gave a n-butanol concentration of 17 g/L with a maximum productivity of 1.1 g/L × h. In terms of productivity and final n-butanol concentration, the results demonstrated that C. pasteurianum DSMZ 525 is well suitable for n-butanol production from mixed substrates of biomass hydrolysate and glycerol and represents an alternative promising production strain.

Knowledge and attitudes towards HIV/AIDS amongst Kuwait University dental students.

The HIV and AIDS have emerged as complex health threats to the world population. As future dentists, it is pertinent that the dental students have sufficient knowledge and a positive approach towards the disease. Accordingly, the aim of this study was to assess the HIV/AIDS related knowledge and attitudes amongst clinical dental students at Kuwait University. A cross-sectional survey was conducted amongst the clinical dental students using a structured questionnaire with 60 questions to examine their knowledge under various categories and 13 questions to examine their attitudes towards the disease. The survey revealed that almost 58% of the respondents demonstrated a high level of knowledge (mean score: 45.23 ± 4.35 SD). Majority of the students (63.6%) expressed negative attitude (mean score: 5.36 ± 2.56 SD). The mean knowledge score of the fifth year dental students was significantly higher (P = 0.022) than that of the final year dental students regarding the knowledge of virus and disease process. However, no significant difference was observed with respect to other knowledge categories. Despite their high level of knowledge, the majority displayed a negative attitude towards HIV/AIDS. Hence, the findings imply that there is a need to address, more clearly, the students' misconceptions and attitudes towards the disease.

Analysis of 3'-Terminal Region of Bean common mosaic virus Strains Infecting Common Bean in India.

Four strains (NL-1, NL-1n, NL-7 and NL-7n) of Bean common mosaic virus (BCMV) prevalent on common bean (Phaseolus vulgaris) in Himachal Pradesh, a north-western Himalayan state of India were compared at the 3' terminal region of the viral genome to elucidate variation and relationship among these strains. 3' terminal region comprising of partial nuclear inclusion b (NIb) (171-233 bp), complete coat protein (CP) (860 bp) and 3' untranslated region (UTR) (208-244 bp) shared 96-98% nucleotide and 96-99% amino acids identity among various strains of BCMV. Multiple alignment as well as cluster dendrograms of the 3' terminal region placed the test isolates in BCMV species of genus Potyvirus. Phylogenetic analysis of complete CP as well as 3' UTR also showed Indian strains to be distinct strains. Sequence homology, multiple alignment and evolutionary divergence of 3' terminal region could not differentiate the pathogenic strain groups, thereby establishing least role of this region in strain characterization of the virus. Comparisons of CP and 3' UTR region of BCMV strains and other members of genus Potyvirus clearly indicated the little utility of 3' terminal region in distinction of virus strains. Implications of coat protein region in viral strain distinction are also discussed.

First Report on Association of Colletotrichum coccodes with Chili Anthracnose in India.

Chili (Capsicum annuum L.) is an important condiment and cash crop grown throughout India, including Andhra Pradesh, Karnataka, Maharashtra, Punjab, Tamilnadu, and Himachal Pradesh. In Himachal Pradesh (HP), a northwestern Himalayan state of India, chilies including sweet pepper occupy an area of 2,447 ha with total production of approximately 31,810 t and productivity of 13.00 t per hectare. In 2007 and 2008, chili- and sweet pepper-growing areas of HP were surveyed for the prevalence of fruit rot/anthracnose disease caused by a complex of Colletotrichum species. Fields infested with disease were randomly sampled and four samples from each location were collected. Disease incidence ranged from 12.5 to 45.0% based on total plants assessed in the field. Symptoms of disease in the field included light brown, sunken lesions containing salmon-colored masses of conidia and microsclerotia on the fruit. Microscopic examination of the diseased samples revealed a variation in morphology of spores from two isolates (Cc 70 and Cc 74) collected from two locations in HP, the Kotkhai area of district Shimla and Shamsher (Ani) locality of district Kullu. Five fruits and ten leaves from five plants of a susceptible local variety were inoculated with a suspension of 5 × 105 conidia/ml of isolates Cc70 and Cc74 using a pin prick method as described by Montri et al. (2). The inoculated fruits and leaves were kept in humid chambers at 25 ± 1°C with 12 h of light. After 48 h, the fruits and leaves were observed daily for the appearance of disease symptoms. Disease symptoms were similar to those of natural infections but with darker lesions. The fungus was recovered from infected fruit on Mathur's medium (glucose 2.80 g, peptone 2.00 g, magnesium sulfate hydrated 1.72 g, potassium dihydrogen orthophosphate 1.23 g, and agar 1.50 g/liter) and initially produced white-to-gray mycelia that became dark brown with age. Setae were present along with production of microsclerotia by the tenth day of culturing. A daily average growth of 8.1 mm was recorded on potato dextrose agar at 25 ± 1°C. Conidia were hyaline, unicellular, aseptate, and fusiform abruptly tapering to each end, and 15.5 to 19.6 µm long and 4.2 to 5.3 µm wide. The fungus was identified as Colletotrichum coccodes based on morphological and cultural traits as per the descriptions of Junior et al. (1). The identity of the isolates was confirmed by amplifying the internal transcribed spacer (ITS) region using primer pair ITS1 and ITS4 (3). The sequences (550 bp) were subjected to a BLAST search with the isolates showing the highest identity to GenBank Accession Nos. GU935878 and EF017205. The sequences have been submitted to GenBank (Accession Nos. HQ264175 and HQ264176). Very few reports exist about the natural occurrence of C. coccodes on Capsicum spp. around the world. To our knowledge, this current report constitutes the first record of this pathogen on Capsicum spp. from the Indian subcontinent. References: (1). H. J. T. Junior et al. Summa Phytopathol. Botucatu 33:418, 2007. (2). P Montri et al. Plant Dis. 93:17, 2009. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Molecular cloning, transcriptional regulation, and differential expression profiling of vitellogenin in two wing-morphs of the brown planthopper, Nilaparvata lugens Stål (Hemiptera: Delphacidae).

The brown planthopper, Nilaparvata lugens, is a serious pest of rice crops throughout Asia and exhibits wing dimorphism, with brachypterous adults having reduced wings and macropterous adults possessing fully developed wings. To understand the reproductive strategies in two wing-morphs of this insect, the transcript encoding the major yolk protein precursor, vitellogenin (Vg), was cloned. The complete mRNA transcript was 6314 bp, which encodes a protein of 2063 residues including an 18-residue putative signal peptide. Analysis of the mature protein revealed two vitellogenin-N (or lipoprotein amino-terminal) domains near the N-terminus and a von Willebrand factor type D domain near the C-terminus. In addition, a highly conserved motif GL/ICG, and a number of cysteine residues were identified near the C-terminus. Northern blot analysis identified a ∼6.8 kb Vg gene transcript that was expressed exclusively in the adult female fat body cells. The expression profile revealed that the Vg gene starts to be expressed earlier (on day 3) in brachypters as compared to macropters where the mRNA transcript was observed on day 4. However, in both morphs, the amount of Vg mRNA increased to reach high levels during vitellogenic periods [from day 4 (in brachypters) and day 5 (in macropters) and onwards]. Reflecting the RNA transcription pattern, the Vg signal was detected by immunoblotting on day 3 and day 4 in haemolymph of brachypterous and macropterous females, respectively, and that was increased every day and remained high during the vitellogenic periods. Furthermore, the topical application of juvenile hormone (JH) III had up-regulated the Vg gene expression suggesting that the Vg gene is regulated by JH in N. lugens. In addition, it was demonstrated by Southern blot analysis that there exists a single copy of the gene in the N. lugens genome. A delayed trend in expression (of both the transcript and the protein) demonstrated by macropterous females in the present studies supports the hypothesis of prereproductive long distance migration in this wing-dimorphic species.

Jatropha curcas: a potential crop for phytoremediation of coal fly ash.

A greenhouse pot experiment was conducted to test the heavy metal phytoremediation capacity of Jatropha curcas from fly ash. Both natural accumulation by J. curcas and chemically enhanced phytoextraction was investigated. Plants were grown on FA and FA amended with fertile garden soil, in presence and absence of chemical chelating agent EDTA at 0.1 g kg(-1) and 0.3 g kg(-1) of soil. EDTA enhanced the uptake of all five elements (Fe, Al, Cr, Cu and Mn) tested. Fe and Mn were retained more in roots while Cu, Al and Cr were translocated more to the shoot. Metal accumulation index indicates that the effect of EDTA at 0.3 g kg(-1) was more pronounced than EDTA at 0.1 g kg(-1) in terms of metal accumulation. Biomass was enhanced up to 37% when FA was amended with GS. Heavy metal uptake was enhanced by 117% in root, 62% in stem, 86% in leaves when EDTA was applied at 0.3 g kg(-1) to FA amended with GS. Study suggest that J. curcas has potential of establishing itself on FA when provided with basic plant nutrients and can also accumulate heavy metals many folds from FA without attenuating plant growth.

Factors associated with severe caries among adults in Kuwait.

AIM: To investigate the relationship between caries etiological factors and occurrence of severe caries among adults in Kuwait. METHODS: Forty-two adult patients with severe caries, attending Kuwait polyclinics, were compared with 36 caries-free subjects in a cross-sectional study. The patients with severe caries were aged 16 years or older and had a minimum of eight open coronal carious cavities, with the involvement of at least one anterior tooth. In addition, their salivary glands had not been compromised by disease or medication. The caries-free subjects had at least 24 standing teeth, and no carious cavities. Salivary flow rates, buffering capacity, frequency of sugar consumption, and oral hygiene index were measured. Mutans streptococci and Lactobacillus counts were determined by the dip-slide method. RESULTS: Fisher's exact test or chi(2) test showed that the patients with severe caries had a significantly higher frequency of sugar consumption, plaque index, Lactobacillus and mutans streptococci counts, as compared with those who were caries-free. In contrast, no significant difference was observed in salivary flow rates or buffering capacity between the two groups of patients. In the multiple logistic regression analysis, only frequency of sugar consumption, oral hygiene, and mutans streptococci count were found to be significantly associated with the occurrence of severe caries. CONCLUSION: Severe caries may occur in Kuwait even among adults whose salivary gland functions have not been compromised.

Psychological distress and associated risk factors in bronchial asthma patients in Kuwait.

CONTEXT: Recent literature shows a high prevalence of psychological distress in bronchial asthma. AIM: To find the extent of psychological distress and associated risk factors in bronchial asthma patients in Kuwait. DESIGN: Case-control study. MATERIALS AND METHODS: In a study at Kuwait's allergy center, 102 patients aged 20-60 years with asthma (67%), asthma with allergic rhinitis (33%) completed a self-administered questionnaire (WHO-Five Well-being Index). A score below 13 was considered as psychological distress; and 13 and above, as normal. An equal number of controls, matched for age, gender, nationality, were also enrolled. STATISTICAL ANALYSIS: The data were analyzed using SPSS software, and proportions were tested with Chi-square or Fisher's test. Odds ratio (OR) with 95% confidence interval (CI) was calculated to quantify the risk factors. RESULTS: A significantly large proportion (69%) of patients were found to be psychologically distressed, compared to 24% among controls (P<0.001, OR=7.5; 95% CI: 4-14). As many as 83.3% of cases, in the younger (20-30 years) age group, were distressed (P<0.044), compared to other age groups. A declining trend in proportion of distressed cases with increasing age was observed (P<0.013). A higher proportion of females (73.8%) and Kuwaitis (71.6%) with distress were observed, both among cases and controls. CONCLUSIONS: We found a high rate of poor well-being and psychological distress in patients suffering from asthma. Young patients and those with relatively short duration of illness, as well as asthmatic females, are more vulnerable to distress and need further psychological evaluation.

Angiotensin-converting enzyme gene polymorphism in Kuwaiti patients with systemic lupus erythematosus.

OBJECTIVE: To investigate the frequency of angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism genotypes in patients with systemic lupus erythematosus (SLE), and to study the correlation between I/D polymorphism of the ACE gene and the clinical manifestations of SLE, especially vascular involvement, lupus nephritis and disease severity. METHODS: The frequency of ACE gene I/D polymorphism genotypes was determined in 92 patients with SLE from Kuwait, and compared to that in 100 ethnically matched healthy controls using the polymerase chain reaction. RESULTS: The distribution of ACE I/D polymorphism and allele frequencies in SLE patients was not significantly different from controls. Further analyses of SLE patients showed that there was a significant association between DD genotype and Raynaud's phenomenon (p=0.008, odd ratio=5.4, 95% confidence interval: 1.6-18.6). However, there was no significant association between the ACE genotype and lupus nephritis or disease severity. CONCLUSION: No difference was found between the distribution of the ACE genotype in SLE patients and the general pop-ulation in Kuwait. However, the presence of the DD genotype may confer susceptibility to the development of vascular morbidity.

Impulse oscillometry: an alternative modality to the conventional pulmonary function test to categorise obstructive pulmonary disorders.

Impulse oscillometry (IOS) was introduced as an alternative modality to the conventional pulmonary function test (cPFT) to test lung mechanics. The objective of this study is to assess the use of IOS as an alternative, or in conjunction with cPFT, to categorise an obstructive respiratory disorder as chronic obstructive pulmonary disease (COPD) or asthma. Patients referred to the PFT laboratory, with different diagnoses of obstructive airways disorders, completed a standardised respiratory questionnaire prior to testing. All recruited subjects completed both modalities of PFT. A total of 146 patients were included. The overall mean age was 51+/-18.4 SD. The majority were non-smokers (68.7%). A standardised diagnosis of asthma was found for 51 subjects, while COPD was diagnosed in 36 subjects and 59 subjects were categorised as normal. The sensitivity of IOS in relation to asthma was 31.3% and 19.6% for cPFT. Among cases of COPD, the cPFT had better sensitivity (47.4%) than IOS (38.95%). The specificity was comparable for IOS and cPFT in relation to asthma and COPD. IOS had better sensitivity (45.8%) in detecting normal subjects than cPFT (28.8%), while specificity was comparable (80.5% and 86.2%, respectively). IOS may replace cPFT where the latter cannot be carried out due to feasibility or lack of cooperation. It can, however, discriminate between diseased and non-diseased subjects.

Septicaemia in scald and flame burns: appraisal of significant differences.

One hundred and sixty burn patients suffering from septicaemia, hospitalized in the Al-Babtain Centre burns unit, Kuwait, between June 1992 and May 2001, were studied. Thirty-two patients (20%) had scalds and 128 (80%) flame burns, thus representing a ratio of 1:4 among septicaemic patients. There were 20 males (62.5%) in the scald group, compared to 73 (57%) with flame burns. Flame burns were significantly higher (p < 0.01) among non-Kuwaiti patients. The mean ages of the scald and flame burn patients were respectively 6.2 and 31.5 yr. The mean total body surface area burn in scalds was 20% and in flame burns 49%, which was significantly higher (p < 0.001). The 34 septicaemic episodes in 32 scald patients and 212 such episodes in 128 flame burn patients showed a significantly higher incidence in the latter group. The majority of septicaemic episodes, in scalds (82.4%) and flame burns (57.6%), were due to gram-positive organisms, mainly methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis. A significantly increased number of episodes were due to S. aureus (p < 0.001) and Enterococcus (p < 0.05) in scald patients. More surgical operations were performed in flame burn patients and survival increased significantly with an increasing number of grafting sessions (p < 0.001). The mean hospital stay in flame burn patients (56 days) was significantly higher than in scald patients (23 days) (p < 0.001). It is significant to record that all the 38 deaths (29.7%) were in flame burn septicaemic patients (p < 0.001). The scald and flame burn patients were quite distinct in their demographic and clinical characteristics. The flame burn patients were more vulnerable to septicaemia, with a high risk of mortality.