Comparative assessment of ISSR, RAPD, and SCoT markers for genetic diversity in Clerodendrum species of North East India.

Clerodendrum belonging to the family of Lamiaceae is used in indigenous systems of medicine to treat various life-threatening diseases. The genus has complex morphological variations which lead to limits in its precise taxonomic classifications. Genetic diversity study could enhance taxonomic authentication and evolutionary relationship among the species of Clerodendrum. In this study, nine species of Clerodendrum collected from different regions of North East India were screened using ISSR, RAPD, and SCoT molecular markers. The markers of ISSR, RAPD, and SCoT generated a total of 79, 126, and 145 amplicons with an average of 6.58, 7.86, and 8.53 amplicon per primer. The polymorphism information contents (PIC) for ISSR, RAPD, and SCoT ranged from 0.28 to 0.37, 0.39 to 0.69, and 0.30 to 0.62 with resolving power (Rp) varying from 5.26 to 11.11, 4.04 to 9.67, and 4.54 to 8.65, respectively. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) based clustering methods grouped 94 genotypes into 6 clusters for ISSR and 3 clusters each for RAPD and SCoT markers. Similarly, population structure-based analysis divided 94 genotypes into 6 populations for ISSR and RAPD and 4 populations for SCoT markers. AMOVA analysis revealed that SCoT markers generated maximum genetic variations within and among genotypes, contrary to ISSR and RAPD markers. Results in this study, suggest that the competence of three markers was relatively the same in genotypes fingerprinting, but SCoT was more efficient in the detection of polymorphism for Clerodendrum species. Further, these results could be integrated in the exploration of diverse Clerodendrum species and germplasm utilization.

Influence of root-knot nematode infestation on antioxidant enzymes, chlorophyll content and growth in Pogostemon cablin (Blanco) Benth.

Plants adapt themselves to overcome adverse environmental conditions, and this involves a plethora of concurrent cellular activities. Physiological experiments or metabolic profiling can quantify this response. Among several diseases of Pogostemon cablin (Blanco) Benth. (Patchouli), root-knot nematode infection caused by Meloidogyne incognita (Kofoid and White) Chitwood causes severe damage to the plant and hence, the oil production. In the present study, we identified M. incognita morphologically and at molecular level using sequenced characterized amplified region marker (SCAR). M. incognita was artificially inoculated at different levels of second stage juveniles (J2) to examine the effect on Patchouli plant growth parameters. Peroxidase and polyphenol oxidase enzyme activity and changes in the total phenol and chlorophyll contents in M. incognita was also evaluated in response to infection. The results have demonstrated that nematode infestation leads to increased peroxidase activities in the leaves of the patchouli plants and thereby, increase in phenolic content as a means of defence against nematode infestation. Chlorophyll content was also found decreased but no changes in polyphenol oxidase enzyme activity.

DNA barcodes for delineating Clerodendrum species of North East India.

The diversified genus of Clerodendrum with its complex evolutionary history leads to taxonomic mystification. Unlike traditional taxonomic methods, DNA barcoding could be a promising tool for the identification and conservation of Clerodendrum species. This study was attempted to develop an efficient barcode locus in Clerodendrum species of North East India. We evaluated four barcode candidates (ITS2, matK, rbcL, ycf1) and its combinations in different Clerodendrum species. The reliability of barcodes to distinguish the species were calculated using genetic pairwise distances, intra- and inter-specific diversity, barcode gap, and phylogenetic tree-based methods. The results exemplify that matK posse's maximum number of variables and parsimony-informative sites (103/100), intra- (0.021 ± 0.001) and inter- (0.086 ± 0.005) specific divergences and species resolution rate (89.1%) followed by ITS2, ycf1, and rbcL. Among the combinatorial locus, ITS2 + matK showed the best species discrimination with distinctive barcode gaps. Therefore, we tentatively suggest that the combination of ITS2 + matK as core barcode for Clerodendrum and converted into quick response (QR) code. Hence, this finding indicates that DNA barcoding could provide consistent resources for species discrimination and resolve taxonomic controversies of the genus as well as set a preliminary assessment toward its biodiversity.

High prevalence of latent tuberculosis infection in patients in end-stage renal disease on hemodialysis: Comparison of QuantiFERON-TB GOLD, ELISPOT, and tuberculin skin test.

BACKGROUND: Individuals with end-stage renal disease (ESRD) are 10- to 25-fold more likely than immunocompetent people to develop active tuberculosis (TB) and are candidates for being treated for latent TB infection (LTBI). However, diagnosis using the tuberculin skin test (TST) is doubly difficult due to cutaneous anergy and cross-reactions with Bacille-Calmette-Guérin (BCG) vaccination. MATERIALS AND METHODS: This was a prospective, doublematched, cohort study in which 32 ESRD patients and 32 age-matched, healthy controls were enrolled. The TST and two new interferon-gamma blood tests, QuantiFERON-TB Gold (QFT-G) and T-SPOT.TB (ELISPOT), were performed. The subjects were followed up 2 years for active TB disease. ELISPOT was done in ESRD patients only. RESULTS: Compared to the healthy controls, a high prevalence of LTBI was found in the ESRD patients by TST (62.5%, 95% confidence interval [CI] 43.7-78.9), QFT-G (40.0%, 95% CI 22.7-59.4), and ELISPOT (46.9%, 95% CI 29.1-65.3). Agreement was moderate (kappa [kappa] = 0.53) for QFT-G and ELISPOT but only slight between TST and QFT-G (kappa = 0.25) and fair between TST and ELISPOT (kappa = 0.32). ESRD (p = 0.03) and diabetes mellitus (p = 0.04) were significant risk factors for QFT-G positivity on the multivariable analysis. The overall rate of active TB was 1.66 cases per 100 person-years (pys), with the rate higher in patients with ESRD (3.53 per 100 pys) and those with positive (3.40 per 100 pys) and indeterminate QFT results (30.16 per 100 pys), although the difference was not statistically significant. Sensitivity, specificity, and positive and negative predictive values of QFT-G for active TB was 100%, 62.1%, 8.3% and 100%. CONCLUSION: This pilot study is the first to compare QFT-G, ELISPOT, and TST in ESRD patients on hemodialysis and demonstrates a high prevalence of LTBI in this population. In our study, the QFT-G was the more accurate method for identifying those truly infected with Mycobacterium tuberculosis, even in BCG-vaccinated individuals.

Efficacy of point-of-entry copper--silver ionisation system in eradicating Legionella pneumophila in a tropical tertiary care hospital: implications for hospitals contaminated with Legionella in both hot and cold water.

A medical centre in Southern Taiwan experienced an outbreak of nosocomial Legionnaires' disease, with the water distribution system thought to be the source of the infection. Even after two superheats and flush, the rate of Legionella positivity in distal sites in hospital wards and intensive care units (ICUs) was 14% and 66%, respectively. Copper-silver ionisation was therefore implemented in an attempt to control Legionella colonisation in both hot- and cold-water systems. Environmental cultures and ion concentration testing were performed to evaluate the efficacy of ionisation. When the system was activated, no significant change in rate of Legionella positivity in the hospital wards (20% vs baseline of 30%) and ICUs (28% vs baseline of 34%) of the test buildings over a three-month period was found, although all Legionella positivity rates were below 30%, an arbitrary target for Legionnaires' disease prevention. When ion concentrations were increased from month 4 to month 7, however, the rate of Legionella positivity decreased significantly to 5% (mean) in hospital wards (P=0.037) and 16% (mean) in ICUs (P=0.037). Legionella positivity was further reduced to 0% in hospital wards and 5% (mean) in ICUs while 50% sites were still positive for Legionella in a control building. Although Legionella was not completely eradicated during the study period, no culture- or urine-confirmed hospital-acquired Legionnaires' disease was reported. Ionisation was effective in controlling Legionella for both hot and cold water, and may be an attractive alternative as a point-of-entry systematic disinfection solution for Legionella.

Endothelin stimulates phospholipase C, Na+/H+ exchange, c-fos expression, and mitogenesis in rat mesangial cells.

A recently described peptide hormone, endothelin, is a potent vasoconstrictor, but it is unclear whether endothelin has other biological actions. These experiments extend the range of biological actions of endothelin to stimulation of mitogenesis. Endothelin at low concentrations (0.1-10 nM) induced mitogenesis by quiescent rat glomerular mesangial cells in culture. Mitogenesis induced by endothelin was accompanied by activation of phospholipase C with increased inositol phosphate turnover and increments of intracellular [Ca2+]. Endothelin also activated Na+/H+ exchange, causing cytosolic alkalinization, and enhanced transcription of the c-fos protooncogene, additional biochemical signals closely linked to proliferation. In addition to being a vasoconstrictor, endothelin thus also functions as a mitogen, presumably through activation of phospholipase C.

Development of an effective and efficient DNA isolation method for Cinnamomum species.

Different species of Cinnamomum are rich in polysaccharide's and secondary metabolites, which hinder the process of DNA extraction. High quality DNA is the pre-requisite for any molecular biology study. In this paper we report a modified method for high quality and quantity of DNA extraction from both lyophilized and non-lyophilized leaf samples. Protocol reported differs from the CTAB procedure by addition of higher concentration of salt and activated charcoal to remove the polysaccharides and polyphenols. Wide utility of the modified protocol was proved by DNA extraction from different woody species and 4 Cinnamomum species. Therefore, this protocol has also been validated in different species of plants containing high levels of polyphenols and polysaccharides. The extracted DNA showed perfect amplification when subjected to RAPD, restriction digestion and amplification with DNA barcoding primers. The DNA extraction protocol is reproducible and can be applied for any plant molecular biology study.

Chlorobutanol, a preservative of desmopressin, inhibits human platelet aggregation and release in vitro.

Therapeutic preparations of desmopressin for parenteral use contain the preservative chlorobutanol (5 mg/ml). We show here that chlorobutanol is a potent inhibitor of platelet aggregation and release. It exhibited a significant inhibitory activity toward several aggregation inducers in a concentration- and time-dependent manner. Thromboxane B2 formation, ATP release, and elevation of cytosolic free calcium caused by collagen, ADP, epinephrine, arachidonic acid and thrombin respectively were markedly inhibited by chlorobutanol. Chlorobutanol had no effect on elastase-treated platelets and its antiplatelet effect could be reversed. It is concluded that the antiplatelet effect of chlorobutanol is mainly due to its inhibition on the arachidonic acid pathway but it is unlikely to have a nonspecific toxic effect. This antiplatelet effect of chlorobutanol suggests that desmopressin, when administered for improving hemostasis, should not contain chlorobutanol as a preservative.

Saphenous vein graft pseudoaneurysm: diagnosis by transesophageal echocardiography and magnetic resonance imaging.

Coronary cineangiography showed probable pseudoaneurysm formation at the origin of an 11-year-old saphenous vein coronary bypass graft. Transesophageal echocardiography and magnetic resonance imaging were helpful in differentiating this from aortic dissection or true aneurysm formation and assisted the surgeon in avoiding perforation of the pseudoaneurysm during sternotomy.

Clinical characteristics of emphysematous pyelonephritis.

A total of 21 patients (20 women and one man) with emphysematous pyelonephritis (EPN), treated in the Kaohsiung Veterans General Hospital during the period from 1991 through 1999 were included in this study. All of the patients were diabetic. The most common symptoms or signs were fever/chills (80%) and costovertebral angle knocking pain (71%). Diagnosis was confirmed by the presence of gas in the parenchyma or paranephric space on plain X-ray of the abdomen or computed tomography. The left kidney (11 cases, 52%) was more frequently affected than the right one (nine cases, 43%), and both kidneys were involved in one case. Obstruction of the corresponding renoureteral unit was found in 19% of the patients, and renal or ureteral stone was found in 23% of the patients. One third of the patients had type I EPN, and two-thirds had type II EPN. Escherichia coli was the most commonly isolated organism, accounting for 61%, 76%, and 47% of isolates from blood, urine, and aspirated pus culture respectively. Prompt control of blood sugar was begun and intravenous antibiotics were given. Drainage was performed in 71% of the patients, and two persons required nephrectomy because of poor control of the infection or complications. Overall survival was 72%. Emphysematous pyelonephritis is a rare, life-threatening, suppurative infection of the renal parenchyma and perirenaL tissues. For successful management of EPN, appropriate medical treatment should be initiated, and immediate nephrectomy or drainage should not be delayed.

Once daily isepamicin treatment in complicated urinary tract infections.

Isepamicin is a new aminoglycoside, derived from gentamicin B, which is more stable than other aminoglycosides against inactivating enzymes, and is less nephrotoxic. We evaluated the efficacy and safety of a once daily isepamicin in the treatment of complicated urinary tract infections (UTIs), as compared with amikacin. During the period May, 1997, to January, 1998, a total of 52 patients with similar demographic and baseline characteristics were enrolled into a prospective, randomized, open-label, single-center trial at the Veterans General Hospital-Kaohsiung. Eleven patients were excluded for protocol violation. The remaining 41 patients were included in the efficacy analysis. Study subjects included 16 men and 25 women, with a mean age of 57.9 (range 18-95) years. Clinical improvement was noted in 100% of patients in both the isepamicin and amikacin group. No statistically significant difference was observed between the 2 groups in regard to the rapidity of defervescence, relief of dysuria and urinary frequency, and clearance of bacteriuria and pyuria. Bacteriological cure rates were 89.4% for the isepamicin group and 100% for the amikacin group. Fifteen of 25 subjects who received isepamicin and 16 of 27 subjects who received amikacin had an adverse effect, all of which were considered to be mild except for one in the amikacin group, who had an adverse event of moderate severity (vomiting). Seven (3 isepamicin and 4 amikacin) adverse events were considered probably or possibly related to the study drug, which included eosinophilia (2 isepamicin), liver function impairment (1 isepamicin, 2 amikacin), renal function impairment (1 amikacin) and flushed face (1 amikacin). However, none of the patients had a life-threatening or severe adverse event that required discontinuation of the drug. These results show that once daily administration of isepamicin is as effective and safe as amikacin in treatment of complicated UTIs.

[Septicemic melioidosis in Southern Taiwan: a case report].

The patient was a 56 year-old man, a resident of Hen-Tsueng Township in Ping-Tung County. He worked as a ranger at Ken-Ting Farm in southern Taiwan, and had been to Thailand for sight-seeing 5 years ago. He came to our outpatient department about one month prior to hospitalization for intermittent fever of one week duration. At that time, complete blood count was within normal limits and a chest roentgenogram was unremarkable. He was given erythromycin without showing any clinical improvement. Two days prior to admission, he noted pain in the left hip. The next day, severe dyspnea developed suddenly and chest x-ray film revealed bilateral nodular lesions. Physical examination on hospitalization revealed an acutely ill and jaundiced male with a temperature of 37.4 degrees C, blood pressure: 110/47 mmHg, pulse rate: 137/min, and respiratory rate: 26/min. There were rales in both lungs, but no lymphadenopathy or organ enlargement. Laboratory study showed WBC: 1,470/mm(3), platelet count: 47,000/mm(3), blood sugar: 226 mg/dL, mildly elevated transaminases and bilirubin, and BUN: 69 mg/dL, Cr: 4.3 mg/dL. Arterial blood gas analysis indicated an acute metabolic acidosis with PaO2 of 32 mmHg. Despite the initial impression of melioidosis and administration of ceftazidime plus gentamicin, his condition rapidly deteriorated and expired 18 hours after admission. Two sets of blood cultures grew Burkholderia pseudomallei. Melioidosis has been called a great imitator of diseases and culture results are frequently misinterpreted. The mortality is high even with suggested therapy with ceftazidime, cotrimoxazole, amoxicillin-clavulanate, chloramphenicol, and/or tetracycline. There has been a total of 10 cases reported in southern Taiwan and 2 of them were clearly indigenous. Melioidosis should be included in the reportable diseases, and its prevalence in Taiwan also should be investigated.

Comparative study of the efficacy and safety of valaciclovir versus acyclovir in the treatment of herpes zoster.

Acyclovir, a specific and selective inhibitor of the replication of Herpesviridae family, has well-documented efficacy and tolerability in the treatment of herpes zoster. Its limited oral bioavailability and short half-life, however, necessitates frequent dosing. Valaciclovir, the l-valyl ester of acyclovir, could be rapidly converted to acyclovir after oral administration, resulting in a three- to five-fold increase in acyclovir bioavailability compared with oral acyclovir in humans. Valaciclovir allows less frequent dosing and maintains the safety profiles of the parent drug. During the period from October 1996 through May 1998, a randomized, prospective study was performed in the Kaohsiung Veterans General Hospital to compare the safety and efficacy of valaciclovir with acyclovir in the treatment of herpes zoster in Taiwanese patients. Patients presenting with herpes zoster within 72 h after the onset of rash were enrolled and randomized to receive one of the following treatments: 1000 mg valaciclovir three times daily for 7 days or acyclovir 800 mg five times daily for 7 days. Patients were followed up for 29 days beginning with the start of therapy. A total of 57 patients were enrolled and randomized to receive valaciclovir (n = 32) or acyclovir (n = 25). Five patients in the valaciclovir group and three in the acyclovir group did not complete the study. The intent-to-treat analysis (57 patients) showed that valaciclovir significantly accelerated the resolution of herpes zoster-associated pain compared with acyclovir; on day 29, the valaciclovir group was 23% superior to the acyclovir group. There was no clinically significant difference in the nature, frequency or severity of adverse events between these two groups, although one and three adverse events were reported in the acyclovir and valaciclovir group, respectively. Thus, we conclude that in the management of herpes zoster, valaciclovir accelerates the resolution of pain and offers a simpler dosing, and maintains the favorable safety profile of acyclovir.

An outbreak of meningitis caused by Angiostrongylus cantonensis in Kaohsiung.

Eight Thai laborers developed meningitis after eating raw snails (Ampullarium canaliculatus) during the period from September 27 to October 6, 1998. The diagnosis of Angiostrongylus cantonensis infection was established in all patients by serologic studies of serum and cerebral spinal fluid (CSF). Clinical manifestations included meningitis, radiculitis and cranial nerve palsy. Symptoms included fever, headache, orbital pain, gastrointestinal upset, hyperesthesia, muscle weakness, skin rash and diplopia. Laboratory abnormalities included peripheral eosinophilia, CSF eosinophilia, transient elevation of liver enzymes and creatinine phosphokinase, elevation of IgE. No space occupying lesions were detected by magnetic resonance imaging of the brain. None of the patients developed severe sequelae during the 6-month follow-up except for occasional headache in one patient. This report also provides evidence that third stage larvae were present in the intermediate host, A. canaliculatus, which the laborers had eaten.

Intramural hematoma of the aorta caused by a penetrating atheromatous ulcer.

Aortic dissection is a life-threatening condition which is difficult to diagnose because of its variable clinical presentation and diverse pathophysiologic manifestations. We present a case in which intramural hematoma of the thoracic aorta occurred in the absence of an intimal flap. Computed tomography and magnetic resonance imaging both showed hemorrhage in the wall of the aorta. Transesophageal echocardiography also revealed a penetrating atherosclerotic ulcer in the transverse aorta as a cause of the hematoma. A high index of clinical suspicion and a combination of diagnostic procedures resulted in proper diagnosis and successful surgical treatment.

Azithromycin in the treatment of pneumonia caused by Chlamydia pneumoniae: report of a case.

We report a 67-year-old male with pneumonia in which Chlamydia pneumoniae was identified by serologic studies as the causative agent. After initial treatment failure with amoxicillin + clavulanic acid, pneumonia was successfully treated with the administration of oral azithromycin, 500 mg per day, for three days. Azithromycin is a new macrolide which has a long half-life and superior action to erythromycin. It provides a new and alternative choice in the treatment of Chlamydia pneumoniae infection in the future.

Comparison of the APACHE II and APACHE III scoring systems in patients with respiratory failure in a medical intensive care unit.

BACKGROUND AND PURPOSE: This retrospective study compared the capability of the Acute Physiology and Chronic Health Evaluation (APACHE) II and APACHE III scoring systems to predict outcome and determined the independent predictors of survival in these scoring systems for patients with respiratory failure in a medical intensive care unit (ICU). MATERIALS AND METHODS: Seven hundred and eight patients with respiratory failure admitted to the medical ICU throughout a 9-year period were studied. Patients with an ICU stay of less than 24 hours, patients under 12 years of age, and burn and surgery patients were excluded. APACHE scores were calculated at 24 hours after admission. Student's t-test was used to compare the total APACHE scores of survivor and non-survivor groups. Multivariate logistic regression analysis was used to determine which variables were predictors of mortality. The discriminative power of APACHE scores to predict in-hospital mortality was studied by the area under the receiver operating characteristic curves of the APACHE II and APACHE III systems, respectively. RESULTS: Both systems showed a significant association between higher scores and higher mortality. The APACHE II system under-predicted the actual hospital mortality rate. The APACHE III systems had a higher discriminative power (area 0.7462) than the APACHE II systems (area 0.6856; p < 0.05). The independent predictors of survival as assessed by APACHE II and III systems were respiratory rate, arterial oxygen pressure, oxygen gradient between alveoli and artery, serum creatinine concentration, and the presence of neurologic abnormalities. CONCLUSIONS: The APACHE III systems has greater discriminative power than the APACHE II systems for predicting in-hospital mortality. The variables of oxygenation, mean artery pressure, respiratory rate, serum creatinine concentration, and Glasgow Coma Scale play important roles in predicting survival for patients with respiratory failure.

Endogenous Escherichia coli endophthalmitis.

Metastatic bacterial endophthalmitis caused by Klebsiella pneumoniae is a unique but well-known phenomenon in Taiwan, where most cases occur in diabetic patients with pyogenic liver abscesses. However, endogenous Escherichia coli endophthalmitis is quite rare. The most common primary site of this infection is the urinary tract and the incidence of concurrent systemic infections, such as abscess formation and infective endocarditis, is high. This illness has a rapidly progressive clinical course associated with a poor prognosis for recovery of visual acuity. This is a report of a case of endogenous E. coli endophthalmitis originating from a renal abscess in a diabetic woman. Despite aggressive local and systemic treatment, evisceration was required to prevent the infection from spreading.

Melioidosis: two indigenous cases in Taiwan.

We report the first two indigenously acquired cases of melioidosis in Taiwan, diagnosed by positive culture and biochemically identified using the ID 32 GN system (BioMerieux Vitek Inc, Hazelwood, MO, USA). The first patient was a 75-year-old Chinese woman who had not travelled abroad since her arrival from mainland China (San-Tung province) 47 years ago. She presented with spontaneous bacterial peritonitis and hepatitis C-related liver cirrhosis with septic shock. Burkholderia pseudomallei (formerly Pseudomonas pseudomallei) was isolated from cultures of both blood and ascites fluid. The second patient, a 70-year-old Chinese man, presented with right lower lobar pneumonia complicated with empyema and septic shock. Blood cultures grew B. pseudomallei. Both patients had underlying diabetes mellitus; one also had liver cirrhosis and chronic renal failure, while the other had a renal stone. The first patient died of refractory septic shock prior to diagnosis. The second patient survived with the use of intravenous ceftazidime for 30 days, followed by oral amoxicillin-clavulanic acid for a further 3 months. These cases serve as a reminder to clinical physicians that melioidosis is now no longer exclusive to patients with a history of travel to endemic areas. A high index of clinical suspicion is required for early diagnosis and treatment in order to reduce the mortality and improve clinical outcome.

Streptococcal toxic shock syndrome in southern Taiwan.

This report outlines our experience with streptococcal toxic shock syndrome (STSS) at the Veterans General Hospital-Kaohsiung during the period October 1990 to November 1993. Group A streptococci were isolated from blood or normally sterile tissue in association with hypotension and multi-organ failure in the eight cases studied. A primary focus of infection was identified in seven cases, including pneumonia (1), septic arthritis (1) and soft-tissue infections (5). The remaining patient suffered from hyperglycemic hyperosmolar non-ketotic coma and Group A streptococcal bacteremia, without an obvious focus of infection. There were four cases of bacteremia. Clinical complications included acute renal failure in all eight cases, disseminated intravascular coagulation in five cases, liver involvement in two cases, adult respiratory distress syndrome in one case and soft-tissue necrosis in five cases. All isolates were sensitive to penicillin, and most patients were treated with intravenous penicillin G, with or without other antibiotics (gentamicin or clindamycin). Of the six patients with soft-tissue infection, two underwent amputation of the infected limb, and one patient underwent sono-guided pigtail drainage of psoas muscle abscess. Three of the patients died. STSS may be uncommon in Taiwan, but it is not rare. Early recognition of STSS (facilitated by Gram stain and culture), prompt debridement and drainage, and adequate antibiotic treatment with penicillin or clindamycin, or both, are necessary for control of such lethal infections.