Diagnosis of Amebic Liver Abscess and Amebic Colitis by Detection of Entamoeba histolytica DNA in Blood,Urine, and Saliva by a Real-Time PCR Assay

The noninvasive diagnosis of amebic liver abscess is challenging, as most patients at the time of diagnosis do not have a concurrent intestinal infection with Entamoeba histolytica. Fecal testing for E. histolytica parasite antigen or DNA is negative in most patients. A real-time PCR assay was evaluated for detection of E. histolytica DNA in blood, urine, and saliva samples from amebic liver abscess as well as amebic colitis patients in Bangladesh. A total of 98 amebic liver abscess and 28 amebic colitis patients and 43 control subjects were examined. The real-time PCR assay detected E. histolytica DNA in 49%, 77%, and 69% of blood, urine, and saliva specimens from the amebic liver abscess patients. For amebic colitis the sensitivity of the real-time PCR assay for detection of E. histolytica DNA in blood, urine, and saliva was 36%, 61%, and 64%, respectively. All blood, urine, and saliva samples from control subjects were negative by the real-time PCR assay for E. histolytica DNA. When the real-time PCR assay results of the urine and saliva specimens were taken together (positive either in urine or saliva), the real-time PCR assay was 97% and 89% sensitive for detection of E. histolytica DNA in liver abscess and intestinal infection, respectively. We conclude that the detection of E. histolytica DNA in saliva and urine could be used as a diagnostic tool for amebic liver abscess.Entamoeba histolytica is a protozoan parasite that causes amebic diarrhea, colitis, and amebic liver abscess (ALA), mostly in developing countries (5, 7, 22, 25). Eighty percent of infected individuals remain asymptomatic carriers, while the other 20% develop clinically overt disease (7, 9, 22, 25). About 50 million symptomatic cases of amebiasis occur worldwide each year, resulting in 40,000 to 100,000 deaths annually (25). Mortality from amebiasis is mainly due to extra-amebic colitis, of which ALA is the most common.It is difficult to differentiate ALA from pyogenic liver abscess or other space-occupying lesions of the liver. Imaging techniques such as ultrasound, computed tomography, and magnetic resonance have excellent sensitivities for the detection of liver abscess arising from any cause, but there are no findings specific for ALA (13). Further complicating the diagnosis is the fact that most patients with an ALA do not have coexistent intestinal infection with E. histolytica (11). Therefore, detection of E. histolytica antigen or DNA in stool samples is not very helpful for the diagnosis of ALA (1, 6, 8, 12).The current means for diagnosis of ALA is the detection of antiamebic antibody by serological tests combined with aspiration of the abscess. The presence of serum antibodies against E. histolytica and the absence of bacteria in the abscess fluid are consistent with an ALA. A drawback of serologic tests is that the serum antibody levels in people from areas of endemicity remain positive for years after infection with E. histolytica (3, 16, 23). Therefore, antiamebic antibodies in the serum may be due to amebiasis in the past, limiting their specificity for the diagnosis of ALA. A further limitation to the current approach to ALA diagnosis is that collection of liver abscess pus is an invasive procedure that requires technical expertise and can be done only in specialized hospitals.Several groups have reported the detection of E. histolytica DNA in liver abscess pus, stool, and other clinical samples by PCR (14, 15, 18, 19, 21, 24, 26). A real-time PCR assay has also been used for detection of E. histolytica DNA in stool and liver abscess pus specimens (2, 10, 20). Real-time PCR has never been used for detection of E. histolytica DNA in urine, saliva, and blood specimens of ALA patients. In this study, we evaluated a real-time PCR assay to detect E. histolytica DNA in blood, urine, and saliva samples of amebic liver abscess and colitis patients in Bangladesh.     

Association between serum cortisol and testosterone levels, opioid therapy, and symptom distress in patients with advanced cancer

ContextPatients with advanced cancer often experience symptoms such as pain, anorexia, and fatigue. Opioid therapy for the management of cancer pain may result in neurohormonal dysfunction that may contribute to a patient’s symptom burden.ObjectivesTo examine the association between serum cortisol and testosterone levels, opioid therapy, and symptom distress in patients with cancer.MethodsA retrospective chart review was performed on 77 consecutive patients with advanced cancer referred for symptoms of fatigue or cachexia. We collected information regarding cortisol levels (am or random), testosterone levels (men only), morphine equivalent daily dose (MEDD), and symptom severity measured by the Edmonton Symptom Assessment Scale. Nonparametric correlation analysis was performed.ResultsThe median age was 63 years (range 24–79), and 62% were men (n = 48). Most patients had gastrointestinal (n = 33, 43%) or thoracic (n = 21, 27%) malignancies and were Caucasian (n = 46, 60%). The median random cortisol level was 19.1 μg/dL (Q1–Q3, 13.4–23.8 [normal, 4.3–22.4]), which correlated with MEDD (Spearman coefficient, 0.25, P = 0.032) and symptoms including pain (0.50, P < 0.001), fatigue (0.29, P = 0.012), nausea (0.34, P = 0.003), depression (0.24, P = 0.032), and anxiety (0.25, P = 0.031). Pain and nausea remained significant after Bonferroni correction. Median morning cortisol level (n = 28) was 20.6 μg/dL (Q1–Q3, 16.6–25.4) and significantly correlated with pain (0.55, P = 0.003) after Bonferroni correction. Patients with a MEDD <30 mg/day had a mean random cortisol level of 16.6 μg/dL, whereas patients with a MEDD ≥30 mg/day had a mean random cortisol level of 20.6 μg/dL (P = 0.01). In 44 male patients with cancer, MEDD was inversely correlated with the total testosterone level (?0.52, P = 0.001).ConclusionIn patients with advanced cancer, elevated random cortisol levels were associated with pain and opioid use, although abnormally low levels of cortisol were found to be infrequent. Patients on higher opioid therapy (MEDD >30) had increased cortisol levels, and male patients had lower testosterone levels. Our study suggests that opioid therapy in patients with advanced cancer may inhibit gonadal function while sparing the adrenal axis. Future studies are needed.     

Synthesis and characterization of biocompatibility of tenorite nanoparticles and potential property against biofilm formation

Aim is to assess the anti-biofilm property of tenorite nanoparticles and to study their suitability as a possible coating material for medical implants. Tenorite (CuO) nanoparticles were synthesized by the optimized thermal decomposition method and characterized using TEM, XRD, FTIR and UV–Vis analysis. Their influence on biofilm formation of microbes was studied by growing multi drug resistant bacterial strains in the presence or absence of these nanoparticles at various concentrations. The cytotoxicity of nanoparticles on mammalian cells was studied at the corresponding concentrations. The nanoparticles were found to be uniformly dispersed, spherical shaped and <50 nm in size. They showed various degrees of anti-biofilm property against clinically isolated, biofilm forming multi drug resistant microorganisms such as Staphylococcus aureus, Pseudomonas fluorescens, Burkholderia mallei, Klebsiella pneumoniae, and Escherichia coli. Furthermore, Hep-2 cells showed excellent viability at tenorite nanoparticles concentration toxic to microbial growth. These results indicate that tenorite nanoparticles may be ideal candidates for being utilized as coating on medical implants in general and dental implants in particular.     

Dermatophytosis in Buffaloes, Cattle and Their Attendants. Dermatophytosen bei Büffeln, Rindern und ihren Wärtern

Summary:  A survey on dermatophytosis was carried out at dairy farms of Habbowal and Punjab Agricultural University (PAU), Ludhiana (India). On direct microscopy and/or culture dermatophytosis was revealed in 107 (0.59%) of 18 099 Murrah buffaloes, 77 (1.56%) of 4 923 cattle and 27 (3.98%) of 678 farm workers. The fungi isolated from various sources in order of decreasing isolation frequency are: Buffaloes: Trichophyton mentagrophytes, T. verrucosum, Microsporum gypseum, Keratinomyces ajelloi, Chrysosporium ; Cattle: T. verrucosum, T. mentagropytes, M. gypseum ; Farm workers: T. rubrum, T. mentagrophytes, M. gypseum, Epidermophyton floccosum, T. violoaceum. The disease occurred throughout the year in all ages, breeds and sexes of animals with a higher prevalence during winter, in young cross-bred male buffaloes and cattle of private dairy farms.
Zusammenfassung:  Auf Milchfarmen der Landwirtschaftlichen Universität Habbowal und Punjab (PAU), Ludhiana (Indien), wurde eine Dermatophytosis-Studie durchgeführt. Mikroskopisch und/oder kulturell wurde an 107 (0.59%) von 18 099 Murrah-Büffeln, an 77 (1.56%) von 4 923 Rindern und an 27 (3.98%) von 678 Landarbeitern eine Dermatophytose festgestellt. Die aus verschiedenen Entnahmebereichen isolierten Pilze waren in abnehmender Häufigkeit: Bei Büffeln: Trichophyton mentagrophytes, T. verrucosum, Microsporum gypseum, Keratinomyces ajelloi, Chrysosporium. Bei Rindern: T. verrucosum, T. mentagrophytes, M. gypseum. Bei Landarbeitern: T. rubrum, T. mentagrophytes, M. gypseum, Epidermophyton floccosum, T. violaceum. Die Krankheit trat das ganze Jahr über in allen Altersstufen, Zuchten und Geschlechtern auf, mit stärkerer Verbreitung im Winter, in jungen, männlichen Hybrid-Büffeln und an Rindern auf privaten Milchfarmen.     

Frequent coinfection with hepatitis B virus strains of distinct genotypes detected by hybridization with type-specific probes immobilized on a solid-phase support

A genotype-specific probes assay (GSPA) was developed for distinguishing the seven genotypes (A-G) of hepatitis B virus (HBV). Nucleotide (nt) sequences corresponding to preS1 region were amplified by PCR with a primer labeled with biotin, and delivered to eight wells on which complementary sequences specific to one or other genotype had been immobilized. Thereafter, hybridization of HBV DNA sequences amplified from the test serum was detected by colorimetry. When 256 sera from HBV carriers in Bangladesh, Cameroon, Japan, South Africa, USA and Uzbekistan were subjected to GSPA, genotypes were concordant with those of ELISA with monoclonal antibodies to epitopes on preS2-region products in 242 (94.6%) of them; 8 sera (3.1%) were not genotypeable by either method. Cloning analysis confirmed the presence of two distinct HBV genotypes in the seven selected sera with coinfection. There were 7 (2.7%) sera with discordant genotyping results between GSPA and ELISA. When HBV DNA clones propagated from these sera were sequenced and analyzed phylogenetically, the genotypes determined by GSPA were verified. Coinfection with HBV strains of two distinct genotypes was identified by GSPA in 28 (10.9%) sera, while it was suggested by ELISA in only 2 (0.8%) sera. The GSPA method would be particularly useful for detecting the coinfection with distinct HBV genotypes of any clinical relevance, which seems to be more frequent than reported previously.