Mindfulness-based therapy for smoking cessation and mental health: a randomised controlled trial.

INTRODUCTION: Effective smoking cessation programmes are essential for assisting smokers in quitting, indirectly lowering mortality and morbidity associated with smoking. Numerous studies have indicated positive outcomes when using mindfulness treatment (MT) to treat psychological or behavioural health issues. Although to date, no study has looked at the effectiveness of online MT for quitting smoking while addressing mental health, particularly among the Asian population. Therefore, this study compares the efficiency of online MT to traditional counselling therapy (CT) in aiding smoking cessation programmes while also addressing mental health. MATERIALS AND METHODS: A randomised control study with a two-group, single-blind design and baseline evaluation was selected. Social media sites were used to advertise for participants, who were then admitted after meeting the requirements. Participants who met the eligibility requirements were randomly split into two groups. Each group received a total of three sessions of online therapy (MT or CT), once every two weeks, as well as one phone call per week as reinforcement. At the beginning and end of the intervention, participants completed questionnaires (1st week and 5th week). Generalized Estimating Equation (GEE) statistical analysis was used to analyse all the variables. RESULTS: The MT group experienced a statistically significant decrease in cigarette consumption (ß: -3.50, 95% Wald CI: - 4.62, -2.39) compared to the CT group over time. Furthermore, the MT group demonstrated significant improvements in their scores for the AAQ-2, anxiety, stress, depression and mindfulness compared to the CT group. CONCLUSION: Online MT is more successful at assisting smokers in lowering their daily cigarette intake and supporting their mental health during the smoking cessation process. Further longitudinal comparisons of the effectiveness of online MT should be undertaken using online platforms in future studies.

Use of the ProSpecT microplate enzyme immunoassay for the detection of pathogenic and non-pathogenic Entamoeba histolytica in faecal specimens.

A commercial microplate enzyme immunoassay (ProSpecT EIA; Alexon Inc., Sunnyvale, CA 94089, USA) was compared with conventional microscopy for the diagnosis of Entamoeba histolytica infection. Using specimens known to be infected, the sensitivity of the ProSpecT EIA was 78% and its specificity was 99%. No cross reaction with other intestinal parasites was observed. The ProSpecT EIA and conventional microscopy (using merthiolate-iodine-formaldehyde direct wet mounts and concentration techniques) were then used to detect E. histolytica infections in 431 patients in a mental hospital in Taiwan. Using single stool specimens, microscopy detected infection in 10.9% of the patients, compared with 16.9% detected by ProSpecT EIA. The latter method was simple and quick, but more expensive, and could be used to complement microscopy if a prompt diagnosis is desired clinically. However, ProSpecT EIA cannot differentiate between pathogenic E. histolytica and non-pathogenic strains (= E. dispar), which limits its usefulness.

Identification of virus-specific vesicles in Giardiavirus-infected Giardia lamblia.

Giardiavirus (GLV), which infects the parasitic protozoan Giardia lamblia, is a nonsegmented double-stranded (ds) ribonucleic acid (RNA) virus. We previously purified two distinct types of related GLV from infected G. lamblia, and showed differential export of one of the viruses from infected cells. In the present study, fractionation of cell lysate was performed, revealing the presence of viruses in the membranous fraction. Distribution of viral antigens in the infected cells was examined by immunocytochemistry. The signal was enriched in certain regions of the cytoplasm, suggesting that a portion of GLV is confined to certain cellular compartments. A significantly reduced signal was also detected in the nuclei. We directly observed the viruses in the infected cells by electron microscopy. Consistent with previous observations, virus-like particles were clearly observed in some membranous vesicles in the cytoplasm at 48 h postinfection, and virus-like particles were again seen in the cytoplasm and then in the nuclei toward the late phase of virus infection. The virus-associated vesicles and some electron-dense nuclear structures were only observed in virus-infected cells, suggesting that virus infection may induce ultrastructural alteration of G. lamblia.

Eye preservation tectonic graft using glycerol-preserved donor cornea.

AIMS: To report the surgical outcome of tectonic graft using glycerol-preserved donor corneas to treat perforated keratitis. METHODS: The medical records were reviewed of all patients treated for perforated keratitis using glycerol-preserved corneas at a single institution between 1 July 2004 and 31 June 2010. The clinical features, precipitating factors, adjuvant therapies, and therapeutic outcomes were analyzed. Success was defined as re-epithelialization of the ocular surface without evisceration. RESULTS: Fourteen eyes from 14 patients (6 male and 8 female) were included. Age ranged from 58 to 84 years (average, 70.71 ± 8.52 years) and the follow-up time ranged from 7 to 56 months (mean, 25.35 ± 16.84 months). The culture results showed five bacterial infections, five cases of fungal keratitis, and one mixed infection; the culture results were negative for three patients. Satisfactory anatomical integrity was obtained in eight grafts (57.14%) that healed with neovascularization. Six grafts (48.85%) showed delayed re-epithelialization and were repaired with conjunctival flaps to maintain ocular surface integrity. Three patients developed secondary glaucoma and received trans-scleral cyclophotocoagulation. Thirteen patients had satisfactory anatomical integrity without evisceration or exenteration, while one patient received evisceration at 39-month follow-up because of intractable glaucoma. CONCLUSIONS: Glycerol-preserved donor corneas combined with anterior vitrectomy with or without conjunctival flaps may be effective substitutes for evisceration surgery in patients with perforated keratitis.

Giardiavirus enters Giardia lamblia WB trophozoite via endocytosis.

As part of our continuing effort in the study of Giardiavirus life cycle, the early events of Giardia lamblia virus (GLV) infection in the trophozoites of G. lamblia WB strain were examined. Electron microscopy showed that GLV particles were initially localized on plasma membrane. As time progresses, GLV was translocated to the peripheral vacuole and then spread to the cytoplasm. Inhibitors of endocytosis such as sodium azide, chloroquine, or ammonium chloride disrupted viral infection when the drug was added simultaneously with the infecting GLV. The inhibitory effect was reduced when sodium azide or chloroquine was added at various intervals after infection. When cells infected for 1 hr were examined by immunofluorescence staining, sodium azide greatly reduced GLV staining signal in general while chloroquine restricted the staining signal to a few bright spots. Significantly more GLV was found in the peripheral vacuoles of chloroquine-treated cells than untreated controls by semiquantitative electron microscopy. In contrast, only a reduced amount of GLV was found in the peripheral vacuoles of sodium azide-treated cells. These results suggest that sodium azide reduces the internalization of infecting GLV, while chloroquine confines the virus in the peripheral vacuoles and, consequently, leads to nonproductive infection. We conclude from these observations that the entry of GLV into the susceptible WB cells in the event of infection is most likely mediated by endocytosis.

Identification of a satellite double-stranded RNA in the parasitic protozoan Trichomonas vaginalis infected with T. vaginalis virus T1.

Co-infection by a 0.5-kb small double-stranded (ds) RNA together with Trichomonas vaginalis virus (TVV) genomic 4.6-kb dsRNA is commonly observed in a number of T. vaginalis isolates. By molecular cloning and primer extension experiments, the 497-bp cDNA sequence of a 0.5-kb dsRNA co-infecting with TVV-T1 in T vagina/is T1 isolate was elucidated. Consistent with the replication cycle of a typical dsRNA virus, a plus-strand viral RNA beginning at +1 of the 0.5-kb dsRNA was identified in infected T. vaginalis T1 cells by primer extension and Northern hybridization studies. The 0.5-kb dsRNA was separately encased in TVV capsids from the viral genomic dsRNA, as shown by protein analysis and electron microscopic examination of viral particles purified by multiple rounds of CsCl gradient centrifugation. The riboprobes transcribed from a cloned cDNA of the 0.5-kb dsRNA exhibited strong hybridization to a small dsRNA in a T vaginalis T9 isolate, which harbors a TVV-T9 distantly related to TVV-T1, but the same probes showed very little hybridization to the viral genomic dsRNA of both TVV-T1 and TVV-T9. Very little sequence homology between the 0.5-kb dsRNA and the 4.6-kb dsRNA in TVV-T1 was found by computer-assisted analysis, suggesting that the small dsRNA in T. vaginalis T1 is not derived from the genome of TVV-T1 or other distantly related T. vaginalis viruses. These results suggest that the small dsRNAs in T vaginalis are satellite RNAs of T. vaginalis virus.

Separation and characterization of two related giardiaviruses in the parasitic protozoan Giardia lamblia.

Giardiavirus encapsidates a 6.2-kb double-stranded (ds) RNA within a capsid that consists of a major 100-kDa capsid protein (p100) and a minor 190-kDa protein (p190). In this study, two nonhomologous 6.2-kb ds RNAs cohabiting in Giardia lamblia trophozoites were found to be separately encapsidated into two distinct virions, one (designated GLV[p100]) whose capsid consists of p100 and p190, and the other (designated GLV[p95]) whose capsid consists of a 95-kDa protein (p95) and a minor p190-equivalent protein. Both types of virions were enriched in the membranous fraction of a lysate from virus-infected G. lamblia cells. Separation of these virions was achieved by CsCl gradient centrifugation following osmotic rupture of the viral particles. By these treatments, the 6.2-kb ds RNA was removed from GLV[p100] whereas that in GLV[p95] remained unchanged, and the two 6.2-kb ds RNAs that had been purified by this protocol displayed differential hybridization properties to viral cDNA probes. Western blotting and peptide mapping experiments show that p100 and p95 were closely related proteins, but each had distinct amino acid sequences. Virus purification and pulse-chase experiments show that GLV[p100] was selectively secreted into the medium whereas GLV[p95] remained within the trophozoites of G. lamblia toward the late phase of cell growth. Secretion of GLV[p100] was not inhibited by Brefeldin A. These findings demonstrate the cohabitation of multiple Giardiavirus species in G. lamblia.

Molecular cloning and characterization of a ras-related gene of ran/tc4/spi1 subfamily in Giardia lamblia.

The significance of Ras-like proteins in the protozoa is relatively unexplored. In this report, a gene encoding a Ras-like nuclear (Ran) protein was identified in Giardia lamblia by a polymerase chain reaction-based cloning strategy. The sequence analyses suggest that the gene was intronless, and had short 5'-untranslated leader sequences in the corresponding mRNA up to -2, -4, or -29 bases upstream of the first initiation codon. The full-length cDNA sequence predicted a protein comprising 226 amino acids, in which the highly conserved functional motifs of the Ras superfamily were all preserved. This protein showed 52% identity to human TC4 and 50% identity to yeast Spi1 proteins, suggesting that it is closely related to the Ran proteins, and it was therefore designated gRan. gRan produced from recombinant Escherichia coli exhibited GTP binding activity by an overlay assay. In good agreement with the predicted size of gRan, a 27-kDa protein was identified in a lysate of G. lamblia by Western blotting using antiserum raised against recombinant gRan. The protein was further localized in both nuclei of G. lamblia by immunofluorescence staining. Recombinant gRan exhibited low affinity for GTP with a Kd value of 16.8 microM. The affinity was enhanced to a Kd value of 2.2 microM in the presence of 10 mM Mg2+. The intrinsic GTPase activity of gRan was observed only in the presence of 10 mM Mg2+ and had an estimated Km of 5.6 microM and a Kcat of 0.33/h. These observations demonstrate the presence of Ras-like proteins in the most primitive eukaryotic cells, G. lamblia, and infer that the Ran protein may play a functional role in the nuclei of this organism.

The course of giardiavirus infection in the Giardia lamblia trophozoites.

The subcellular distribution of Giardia lamblia virus RNA in infected G. lamblia trophozoites was examined by in situ hybridization using biotinylated DNA probe and riboprobe. In G. lamblia Portland I strain, which is chronically infected by G. lamblia viruses, the viral RNA was detected in the cytoplasm as well as in the twin nuclei. When riboprobe was used to examine the course of virus infection in WB strain, accumulation of viral RNA was detected only in the cytoplasm prior to the first 72 hr of infection. Using DNA probe, further accumulation of viral RNA in increasing number of cells occurred after the 72nd hr of infection, with the RNA found in both the cytoplasm and nuclei. Eventually, the cell nuclei showed damaged morphology that deteriorated rapidly toward the final stage of infection. These observations indicate that early phase of viral RNA replication may take place in the cytoplasm of infected G. lamblia, but the nuclei are also involved during the late phase of viral replication.

Human babesiosis in Taiwan: asymptomatic infection with a Babesia microti-like organism in a Taiwanese woman.

An asymptomatic Babesia infection was confirmed by laboratory diagnoses. The intraerythrocytic protozoan (designed TW1) isolated from a 51-year-old Taiwanese woman appeared to be morphologically consistent with small-form piroplasm, and measurements indicated that it had a body size of 1.5 to 2.5 microm in diameter. The typical features of ring, binary, and tetrad forms were observed in Giemsa-stained thin blood smears. A persistent and low-grade parasitemia was established after hamster inoculation. Indirect immunofluorescent-antibody reactivities indicate that this strain (TW1) of Babesia was serologically related to, but not identical to, the Babesia species (B. microti) that infects rodents. Antibody titers in the patient's sera combined with the clinical symptoms suggested that the present case was a chronic and subclinical babesial infection. A neighborhood human serologic survey indicated that the infection may have been acquired accidentally from an infected rodent and localized within the same family. Indeed, rodents from areas around the neighborhood were trapped, and a high prevalence (83%) of babesial infection was observed. The possible vector responsible for the transmission remains to be identified.