Outcome of Treatment in Verrucous Carcinoma of Oral Cavity: A Tertiary Rural Hospital Experience.

Verrucous carcinoma of oral cavity is a highly well differentiated variant of squamous cell carcinoma with a low potential for invasion and metastasis. It is prevalent in the tobacco quid chewing population in our region. In this observational study, we reviewed the medical case records of 58 patients treated for oral verrucous carcinoma staged T2 to T4a. All patients underwent wide excision of tumour which included marginal mandibulectomy in 22 and hemimandibulectomy in 23 patients along with neck dissection saving the accessory nerve and internal jugular vein. 5 patients were found to have bone involvement along the alveolar sockets. 11 patients had other associated premalignant lesions in oral cavity. Only 2 patients had lymph node metastasis without extra nodal spread in submandibular region. With a mean follow up of 6 years and minimum follow up of 1 year, 3 patients had local recurrence. All these 3 patients had bone involvement and 2 of them had lymph node metastasis on histopathological examination. 3 patients who had associated premalignant lesions developed second primary in oral cavity after 3 years. In our experience, verrucous carcinoma has good prognosis when treated by surgery. Bone involvement along alveolar sockets and associated oral premalignant lesions adversely affect the outcome. There was no difference in the outcome between selective and modified radical neck dissection. Therefore selective neck dissection (supraomohyoid) would be adequate in treating these patients. Adjuvant radiotherapy can be reserved for T4a lesions or for positive margins.

Outcome of Treatment in Locally Advanced Upper Alveolar Carcinoma Extending to Infra-temporal Fossa.

Squamous cell carcinoma of oral cavity has a high prevalence in our region. Upper alveolar cancers are uncommon but present with locally advanced disease extending to infra temporal fossa. The outcome of treatment in these tumors is poor. Surgery followed by adjuvant therapy is the mainstay of treatment. We are presenting a retrospective analysis of outcome of treatment in 20 patients with locally advanced (T4b) upper alveolar carcinoma treated by infra-structure maxillectomy with compartment resection of infra-temporal fossa and neck dissection followed by adjuvant radiotherapy or radiotherapy with chemotherapy. Study period was from 2013 to 2018. Minimum follow up was 12 months and mean follow up 30 months. 12 patients are alive and disease free, 6 patients had local recurrence and 2 patients had regional recurrence with one having pulmonary metastasis. We observed that positive or close margins (< 5 mm after formalin fixation) predisposed to early recurrence. Erosion of pterygoid plates was a poor prognostic factor. Cervical lymph node metastasis occurred late in disease, but carried poor prognosis. Compartment resection of infra temporal fossa gave better outcome compared to other studies which reported outcome of treatment in upper alveolar cancers.

Impact of isolation precautions on quality of life: a meta-analysis.

Impact of isolation precautions on psychological wellbeing of patients has yet to be fully quantified. To assess the impact of isolation precautions on patients' health-related quality of life and depression or anxiety scales and estimate per day cost of anxiety and depression. Literature pertaining to impact of isolation precautions was searched on EMBASE and PubMed databases and Google Scholar. A two-step independent screening of the articles was performed. Articles that compared isolated and non-isolated patients using different quality of life and psychological burden scales were included. A meta-analysis was conducted using the Hospital Anxiety and Depression Scales (HADS-A and HADS-D). Psychological burden measures from selected literature were presented in a graph as effect sizes. Per day cost of anxiety and depression was estimated using pooled mean difference from meta-analysis. Out of 106 articles, 94 were excluded due to inclusion criteria, leaving 12 for full text review. After review of full text of the articles, seven articles were shortlisted for empirical analysis and four out of these seven for meta-analysis. The pooled mean difference estimates for HADS-A was -1.4 (P=0.15) and that for HADS-D was -1.85 (P=0.09). In the empirical analysis of psychological burden scales, the effect in all studies except one was negative. Results from meta-analysis and empirical analysis of psychological burden implied that isolated patients are worse off in general. The implied estimated per day cost of anxiety and depression in terms of quality-adjusted life years (QALYs) is approximately US$10.

Lethal Midline Granuloma: An Enigma in Diagnosis-A Rare Case Report.

Lethal midline granuloma is a rare disease, characterised by progressive unrelenting necrosis and mutilation of nose, midline facial tissues and other respiratory passages. This particular type of disease is heterogeneous in its pathogenesis, non-specificity of symptoms obscures timely and correct diagnosis and is responsible for delay in of treatment which can be detrimental as this disease calls for immediate intervention. We present a case report of 60 year old female who gave short one-month history of clinical symptoms.

Characterization of Fusobacterium necrophorum subsp. necrophorum outer membrane proteins.

Liver abscesses are of major economic importance to the cattle industry. These are mainly associated with the presence of Fusobacterium necrophorum, a non-spore forming and Gram-negative anaerobe. There are two main subspecies, F. necrophorum subspecies necrophorum and subsp. funduliforme, and they differ molecularly, morphologically, biochemically and in virulence. Previous studies have shown that the outer membrane proteins (OMP) of F. necrophorum subsp. necrophorum are important for its successful binding to immobilized bovine adrenal gland capillary endothelial (EJG) cells. In this study, a 42.4 kDa OMP of F. necrophorum subsp. necrophorum with the highest binding capacity to EJG cells was characterized. The gene was cloned into pFLAG-CTS vector and the proteins were subsequently expressed on the surface of E. coli BL21 DE3 cells. When E. coli carrying the recombinant plasmid (SM 2013) was induced using IPTG, there was significant enhancement in the binding to immobilized EJG cells compared to both uninduced SM 2013 and the E. coli carrying control vector only. When fixed EJG cells were incubated with purified native OMP, SM 2013 showed lowered levels of binding, compared to the uninduced SM 2013 and the E. coli carrying control vector only. Pre-incubation of induced SM 2013 with polyclonal antibodies made against the OMP reduced the binding to immobilized EJG cells to uninduced SM 2013 levels. This gain of function by recombinant E. coli confirms the ability of this protein to act as an adhesion to help binding of F. necrophorum subsp. necrophorum to host cells.

Conditional reprogramming of pediatric airway epithelial cells: A new human model to investigate early-life respiratory disorders.

BACKGROUND: Airway epithelial cells (AEC) are quite difficult to access in newborns and infants. It is critically important to develop robust life-extended models to conduct translational studies in this age group. We propose the use of a recently described cell culture technology (conditionally reprogrammed cells-CRC) to generate continuous primary cell cultures from nasal and bronchial AEC of young children. METHODS: We collected nasal and/or bronchial AEC from a total of 23 subjects of different ages including newborns/infants/toddlers (0-2 years; N = 9), school-age children (4-11 years; N = 6), and a group of adolescent/adult donors (N = 8). For CRC generation, we used conditioned medium from mitotically inactivated 3T3 fibroblasts and Rho-associated kinase (ROCK) inhibitor (Y-27632). Antiviral immune responses were studied using 25 key antiviral genes and protein production of type III epithelial interferon (IFN λ1) after double-stranded (ds) RNA exposure. RESULTS: CRC derived from primary AEC of neonates/infants and young children exhibited: (i) augmented proliferative capacity and life extension, (ii) preserved airway epithelial phenotype after multiple passages, (iii) robust immune responses characterized by the expression of innate antiviral genes and parallel nasal/bronchial production of IFN λ1 after exposure to dsRNA, and (iv) induction of airway epithelial inflammatory and remodeling responses to dsRNA (eg, CXCL8 and MMP9). CONCLUSION: Conditional reprogramming of AEC from young children is a feasible and powerful translational approach to investigate early-life airway epithelial immune responses in humans.

Endotypes of difficult-to-control asthma in inner-city African American children.

African Americans have higher rates of asthma prevalence, morbidity, and mortality in comparison with other racial groups. We sought to characterize endotypes of childhood asthma severity in African American patients in an inner-city pediatric asthma population. Baseline blood neutrophils, blood eosinophils, and 38 serum cytokine levels were measured in a sample of 235 asthmatic children (6-17 years) enrolled in the NIAID (National Institute of Allergy and Infectious Diseases)-sponsored Asthma Phenotypes in the Inner City (APIC) study (ICAC (Inner City Asthma Consortium)-19). Cytokines were quantified using a MILLIPLEX panel and analyzed on a Luminex analyzer. Patients were classified as Easy-to-Control or Difficult-to-Control based on the required dose of controller medications over one year of prospective management. A multivariate variable selection procedure was used to select cytokines associated with Difficult-to-Control versus Easy-to-Control asthma, adjusting for age, sex, blood eosinophils, and blood neutrophils. In inner-city African American children, 12 cytokines were significant predictors of Difficult-to-Control asthma (n = 235). CXCL-1, IL-5, IL-8, and IL-17A were positively associated with Difficult-to-Control asthma, while IL-4 and IL-13 were positively associated with Easy-to-Control asthma. Using likelihood ratio testing, it was observed that in addition to blood eosinophils and neutrophils, serum cytokines improved the fit of the model. In an inner-city pediatric population, serum cytokines significantly contributed to the definition of Difficult-to-Control asthma endotypes in African American children. Mixed responses characterized by TH2 (IL-5) and TH17-associated cytokines were associated with Difficult-to-Control asthma. Collectively, these data may contribute to risk stratification of Difficult-to-Control asthma in the African American population.

Comparison of Mannheimia haemolytica isolates from an outbreak of bovine respiratory disease.

The objective of this study was to determine the clonal relatedness of Mannheimia haemolytica isolates responsible for an outbreak of bovine respiratory disease in a commercial feedlot. The isolates were obtained from the lungs of 21 calves with fatal pneumonia that were part of a group of 206 total calves. All isolates were serotyped and analyzed by pulsed-field gel electrophoresis (PFGE) and for antibiotic sensitivity patterns. ELISA and immunoblotting assays were performed to compare serum antibody levels to M. haemolytica antigens in calves with fatal pneumonia to those calves that survived the outbreak. Isolates were categorized into 14 different PFGE groups based on 90% similarity. Two Group D isolates (1 and 6), and 3 Group H isolates (14, 15, and 16) were characterized as 100% similar. Antimicrobial susceptibility profiles defined 8 groups based on differences in patterns of resistance between isolates. The two 100% similar isolates from PFGE Group D were both in susceptibility Group 1. All but isolate 14 from PFGE Group H (3, 15, 16, and 19) were in susceptibility Group 4a. Serum antibody levels to M. haemolytica antigens in the dead calves were not different than the antibody levels in the 185 calves that survived the outbreak. Immunoblots of selected isolates from each of the PFGE groups demonstrated only minimal differences in antigenic profiles between strains when reacted with serum from calves that either died from or survived the outbreak. Based on the characteristics of these isolates, multiple strains of M. haemolytica were responsible for fatal pneumonia during this outbreak.

Multicenter evaluation of the BD max enteric bacterial panel PCR assay for rapid detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga toxin 1 and 2 genes.

Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens.

Morbidity and mortality of diabetes with surgery.

The prevalence of Type 2 diabetes mellitus (T2DM) has increased; as a result the number of patients with T2DM undergoing surgical procedures has also increased. This population is at high risk of macrovascular (cardiovascular disease, peripheral vascular disease) or microvascular (retinopathy, nephropathy or neuropathy) complications, both increasing their perioperative morbidity and mortality. Diabetes patients are more at risk of poor wound healing, respiratory infection, myocardial infarction, admission to intensive care, and increased hospital length of stay. This leads to increased inpatient costs. The outcome of perioperative glycaemia management remains a significant clinical problem without a universally accepted solution. The majority of evidence on morbidity and mortality of T2DM patients undergoing surgery comes from the setting of cardiac surgery; there was less evidence on noncardiac surgery and bariatric surgery. Bariatric surgery is increasingly performed in patients with severe obesity complicated by T2DM, but is distinguished from general surgery as it immediately improves the glucose homeostasis postoperatively. The improvements in glycaemia are thought to be independent of weight loss and this requires different postoperative management. Patients usually have to follow specific preoperative diets which lead to improvement in glycaemia immediately before surgery. Here we review the available data on the mortality and morbidity of patients with T2DM who underwent elective surgery (cardiac, non-cardiac and bariatric surgery) and the current knowledge of the impact that preoperative, intraoperative and postoperative glycaemic management has on operative outcomes.

La prevalencia de la diabetes mellitus tipo 2 (DM2) ha incrementado en los últimos años, y como resultado, el número de pacientes con DM2 sometidos a procedimientos quirúrgicos también ha aumentado. Esta población posee un alto riesgo de complicaciones macrovasculares (enfermedad cardiovascular, enfermedad vascular periférica) o microvasculares (retinopatía, nefropatía o neuropatía), ambos incrementan tanto la mortalidad como la morbilidad perioperatoria de estos pacientes. Los pacientes con diabetes tienen un mayor riesgo de una mala cicatrización de las heridas, infección respiratoria, infarto de miocardio, ingreso en la UCI y mayor duración de la estancia hospitalaria. Todo esto incrementa los costes de tratamiento de este tipo de pacientes. El control de la glucemia perioperatoria sigue siendo un importante problema clínico sin una solución universalmente aceptada. La mayoría de los conocimientos sobre la morbilidad y mortalidad de los pacientes con DM2 sometidos a cirugía proviene de la de la cirugía cardíaca, y algunos, aunque menos, de la cirugía no cardiaca y cirugía bariátrica. La cirugía bariátrica se realiza cada vez más en pacientes con obesidad mórbida complicado con diabetes tipo 2, y se diferencia de la cirugía general en que inmediatamente mejora la homeostasis de la glucosa tras la operación. Las mejoras en el control de la glucemia parecen ser independientes de la pérdida de peso y esto requiere un manejo postoperatorio diferente. Los pacientes por lo general tienen que seguir dietas específicas preoperatorias que conducen a la mejora de la glucemia inmediatamente antes de la cirugía. En este artículos revisamos los datos disponibles sobre la mortalidad y la morbilidad de los pacientes con diabetes tipo 2 sometidos a cirugía (cirugía cardíaca, no cardíaco y bariátrica) y el conocimiento actual de los efectos preoperatorios, intraoperatorios y postoperatorios que el control de la glucemia tiene sobre los resultados operatorios.

Morbidity and mortality of diabetes with surgery / Morbi-mortalidad en pacientes diabéticos tipo 2 tras cirugía electiva

The prevalence of Type 2 diabetes mellitus (T2DM) has increased; as a result the number of patients with T2DM undergoing surgical procedures has also increased. This population is at high risk of macrovascular (cardiovascular disease, peripheral vascular disease) or microvascular (retinopathy, nephropathy or neuropathy) complications, both increasing their perioperative morbidity and mortality. Diabetes patients are more at risk of poor wound healing, respiratory infection, myocardial infarction, admission to intensive care, and increased hospital length of stay. This leads to increased inpatient costs. The outcome of perioperative glycaemia management remains a significant clinical problem without a universally accepted solution. The majority of evidence on morbidity and mortality of T2DM patients undergoing surgery comes from the setting of cardiac surgery; there was less evidence on non-cardiac surgery and bariatric surgery. Bariatric surgery is increasingly performed in patients with severe obesity complicated by T2DM, but is distinguished from general surgery as it immediately improves the glucose homeostasis postoperatively. The improvements in glycaemia are thought to be independent of weight loss and this requires different postoperative management. Patients usually have to follow specific preoperative diets which lead to improvement in glycaemia immediately before surgery. Here we review the available data on the mortality and morbidity of patients with T2DM who underwent elective surgery (cardiac, non-cardiac and bariatric surgery) and the current knowledge of the impact that preoperative, intraoperative and postoperative glycaemic management has on operative outcomes (AU)

La prevalencia de la diabetes mellitus tipo 2 (DM2) ha incrementado en los últimos años, y como resultado, el número de pacientes con DM2 sometidos a procedimientos quirúrgicos también ha aumentado. Esta población posee un alto riesgo de complicaciones macrovasculares (enfermedad cardiovascular, enfermedad vascular periférica) o microvasculares (retinopatía, nefropatía o neuropatía), ambos incrementan tanto la mortalidad como la morbilidad perioperatoria de estos pacientes. Los pacientes con diabetes tienen un mayor riesgo de una mala cicatrización de las heridas, infección respiratoria, infarto de miocardio, ingreso en la UCI y mayor duración de la estancia hospitalaria. Todo esto incrementa los costes de tratamiento de este tipo de pacientes. El control de la glucemia perioperatoria sigue siendo un importante problema clínico sin una solución universalmente aceptada. La mayoría de los conocimientos sobre la morbilidad y mortalidad de los pacientes con DM2 sometidos a cirugía proviene de la de la cirugía cardíaca, y algunos, aunque menos, de la cirugía no cardiaca y cirugía bariátrica. La cirugía bariátrica se realiza cada vez más en pacientes con obesidad mórbida complicado con diabetes tipo 2, y se diferencia de la cirugía general en que inmediatamente mejora la homeostasis de la glucosa tras la operación. Las mejoras en el control de la glucemia parecen ser independientes de la pérdida de peso y esto requiere un manejo postoperatorio diferente. Los pacientes por lo general tienen que seguir dietas específicas preoperatorias que conducen a la mejora de la glucemia inmediatamente antes de la cirugía. En este artículos revisamos los datos disponibles sobre la mortalidad y la morbilidad de los pacientes con diabetes tipo 2 sometidos a cirugía (cirugía cardíaca, no cardíaco y bariátrica) y el conocimiento actual de los efectos preoperatorios, intraoperatorios y postoperatorios que el control de la glucemia tiene sobre los resultados operatorios (AU)

A new twist on an old problem: a case of pediatric meningitis caused by multidrug-resistant Streptococcus pneumoniae serotype 19A.

Recent studies show that multidrug-resistant Streptococcus pneumoniae serotype 19A continues to emerge as a cause of invasive pneumococcal disease after the introduction of Prevnar. We report a case of multidrug-resistant S. pneumoniae serotype 19A meningitis successfully treated with vancomycin and levofloxacin. This case reinforces the need for the empiric use of vancomycin in meningitis and the need for alternative treatments.

Pelvic Ewing's sarcoma: a review from Scottish Bone Tumour Registry.

PURPOSE: To review various treatment modalities on outcome of patients with pelvic Ewing's sarcoma. METHODS: Between January 1948 and December 2004, 18 male and 15 female patients aged 3 to 48 (mean, 20) years with pelvic Ewing's sarcoma were retrospectively reviewed from the Scottish Bone Tumour Registry. RESULTS: The commonest site involved was the ilium. The main presenting symptom was pain, followed by swelling and restricted hip movements. Treatments included surgery, radiotherapy, chemotherapy, or any of them in combination. Patients who had no metastasis at presentation (p<0.01) and underwent chemotherapy with ifosfamide (p<0.01) had significantly longer survival. Age (p=0.09), gender (p=0.95), delay in presentation (p=0.31), tumour site (p=0.9), surgery (p=0.73), and radiotherapy (p=0.23) were not predictive of survival in the univariate analysis. The 5-year event-free survival rate in patients with no metastasis at presentation was 35%. CONCLUSION: Intense multi-agent neo-adjuvant chemotherapy including ifosfamide followed by wide excision and postoperative radiation are recommended. Local therapy should not take precedence over or interfere with systemic chemotherapy.

Rapid detection of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), the pathogenic agents of white tail disease of Macrobrachium rosenbergii (De Man), by loop-mediated isothermal amplification.

A loop-mediated isothermal amplification (LAMP) procedure is described for rapid diagnosis of white tail disease, a viral disease caused by Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV), in the giant freshwater prawn, Macrobrachium rosenbergii. This method was more sensitive than conventional RT-PCR for detecting the two viruses. A set of four primers, two outer and two inner, were designed for MrNV detection. An additional pair of loop primers was also used in an accelerated LAMP reaction for detection of XSV. Time and temperature conditions were optimized for detection of the two viruses. The LAMP reaction is highly suited for disease diagnosis in developing countries as amplification of DNA can be detected without the use of agarose gel electrophoresis, by the production of whitish precipitate of magnesium pyrophosphate as a by-product.

Gross signs and histopathology of branchiostegal blister disease (balloon disease): an idiopathic disease of farmed Macrobrachium rosenbergii (De Man).

The giant freshwater prawn, Macrobrachium rosenbergii, is facing increased threat due to disease as its culture becomes more widespread. A disease characterized by the swelling of the branchiostegal region and deformities of the appendages, named balloon disease by farmers, has caused considerable economic loss in the Nellore region of Andhra Pradesh, India. Clinical signs of diseased animals include a voluminous hypertrophy of some gill filaments and the inner area of the branchiostegite. By histology, hypertrophied areas at the level of the gill filaments or branchiostegite had an identical structure corresponding to a large cyst filled with a fluid containing a few free haemocytes, limited on one side by the cuticle and on the other by the subcuticular epithelial layer. Analysis of the diseased prawns did not reveal any pathogenic agent leading us to conclude that the disease is idiopathic, probably due to suboptimal water quality conditions.

Plasmodium falciparum malaria in Laos: chloroquine treatment outcome and predictive value of molecular markers.

A 28-day treatment trial was undertaken, to determine the efficacy of chloroquine in Laos and to assess the predictive value of molecular markers (cg2, pfmdr1, and pfcrt) that were previously linked to chloroquine resistance. In total, 522 febrile patients were screened for falciparum malaria by rapid diagnostic assays. Of 81 patients (15.5% prevalence) who were positive by the assays and microscopy, 48 were eligible to participate in the 28-day trial. Nine patients defaulted. Chloroquine cured 54% (95% confidence interval, 45.8-61.8) of falciparum-infected patients. Of 18 (46%) patients with treatment failure, 13 (72%) experienced high-grade resistance. Polymorphisms in cg2 and the N86Y mutation in PfMDR1 were not predictive of treatment outcome. A mutation in PfCRT (K76T) was perfectly associated with in vivo chloroquine resistance. However, K76T was also present in in vivo-sensitive isolates, which suggests that the presence of this mutation was necessary, but not sufficient, to predict in vivo outcome in this cohort.

Entamoeba dispar: molecular characterization of the galactose/N-acetyl-d-galactosamine lectin.

Amebiasis contributes to approximately 50 million cases of life-threatening dysentery worldwide. Comparison of the lectins from Entamoeba histolytica (pathogenic) and Entamoeba dispar (nonpathogenic) was undertaken to elucidate the differential roles of this molecule in invasion versus colonization. Surface lectin was less abundant on axenic E. dispar than on axenic E. histolytica, commensurate with differences in lectin (heavy and light subunits) RNA when assessed by semiquantitative RT-PCR. The 1G7 epitope, which falls within the immunodominant and immunoprotective cysteine-rich region (480-900), was absent on axenic E. dispar. Indirect immunofluorescence, transient transection of COS7, and immunoprecipitation demonstrated that the 1G7 epitope was conserved in the nonpathogenic lectin homologue but not exposed on live E. dispar trophozoites. Hgl2 (E. histolytica) and Dhgl2 (E. dispar) lectin homologues demonstrated comparable high-affinity binding to multivalent GalNAc(19) BSA. These data provide evidence for relative gene and conformational regulation of the E.dispar lectin.

The performance and utility of rapid diagnostic assays for Plasmodium falciparum malaria in a field setting in the Lao People's Democratic Republic.

Rapid diagnostic assays for malaria have the potential to improve the management and control of the disease in developing countries. The objectives of the present study were to evaluate, in a field setting, the performance of several such assays for Plasmodium falciparum infection and to examine the usefulness of these assays in identifying subjects for treatment trials in rural field sites. Residents of 12 villages in Laos who presented with fever were eligible for inclusion. Blood was collected by fingerprick for a dipstick assay, developed by the Program for Appropriate Technology in Health (PATH), performed and interpreted in the field by local healthcare workers. Compared with 'blinded' reference microscopy (N =196), the sensitivity and specificity of the PATH assay were 96.2% and 93.0%, respectively. Two rapid diagnostic assays (PATH and OptiMAL) were also performed on the subset of subjects eligible to participate in an in-vivo treatment trial (N = 97), and the results again compared with those of 'blinded' reference microscopy. In this subset, a subject was considered a 'true positive' if found positive by microscopy or the alternate rapid assay. Using this modified reference standard, the sensitivity and specificity of the PATH assay were 96.7% and 94.4%, and those of the OptiMAL assay were 91.8% and 100%, respectively. Both of the rapid assays tested therefore appear suitable for use in rural field settings by local healthcare providers and can accurately identify participants for treatment trials.

Molecular markers for chloroquine-resistant Plasmodium falciparum malaria in Thailand and Laos.

Chloroquine-resistant Plasmodium falciparum is well documented in Thailand. Laos, however, continues to use chloroquine (CQ) as the first-line therapy for the treatment of P. falciparum malaria. The objective of the present study was to determine the prevalence, in these two areas, of the cg2, pfmdr1 and pfcrt allelic types that have previously been associated with CQ resistance. Isolates of P. falciparum were collected from participants in ongoing treatment studies conducted in Thailand (near the Thai-Cambodian border) and in Laos (Vang Vieng district). The pfmdr1 and pfcrt alleles were characterized by PCR-RFLP and mutations in cg2 were characterized by PCR and single-stranded-conformation-polymorphism (SSCP) electrophoresis. Eight (32%) of the 25 Laotian isolates but only one (4%) of the 25 Thai isolates were found to contain the pfmdr1 mutation N86Y (P = 0.02). In contrast, the cg2 polymorphisms previously associated with CQ resistance were present in only 10 of the isolates from Laos but 24 of those from Thailand (40% v. 96%; P < 0.001). All the samples from both countries contained the pfcrt K76T mutant allele reported to confer resistance to CQ. The results may indicate that drug pressure for the maintenance of the pfmdr1 and cg2 alleles varies in intensity in the Thai and Laotian study areas, probably reflecting differences in the national malaria-treatment policies of Thailand and Laos.