Draft Genome Sequence of Sphingobacterium sp. Strain HMA12, Which Encodes Endo-ß-N-Acetylglucosaminidases and Can Specifically Hydrolyze Fucose-Containing Oligosaccharides.

The genome sequence of the soil bacterium Sphingobacterium sp. strain HMA12, the culture supernatant of which exhibited endo-ß-N-acetylglucosaminidase (ENGase) activity, was examined for ENGase-encoding genes. Here, we report the characterization of new genes of ENGases, obtained by whole-genome shotgun sequencing, that are capable of specifically hydrolyzing fucose-containing oligosaccharides.

Unfinished Business in Families of Terminally Ill With Cancer Patients.

CONTEXT: Unfinished business often causes psychological issues after bereavement. Providing care for families of terminally ill patients with cancer to prevent unfinished business is important. OBJECTIVES: To clarify the prevalence and types of unfinished business in families of end-of-life patients with cancer admitted to palliative care units (PCUs), explore depression and grief associated with unfinished business, and explore the factors affecting unfinished business. METHODS: We conducted a cross-sectional, anonymous, self-report questionnaire survey with 967 bereaved families of patients with cancer admitted to PCUs. The questionnaire assessed the presence or the absence of unfinished business, content of unfinished business, depression, grief, process of preparedness, condition of the family and patient, and the degree of involvement of health care professionals. RESULTS: Questionnaires were sent to 967 families, and 73.0% responded. In total, 26.0% of families had some unfinished business, with improvement of the patient-family relationship being a common type of unfinished business. Families with unfinished business had significantly higher depression and grief scores after bereavement compared with those without. Factors that influenced the presence or the absence of unfinished business were preparedness for the patient's death (P = 0.001), discussion between the patient and family about the disease trajectory and way to spend daily life (P < 0.001), good patient-family relationship (P = 0.011), and family and health care professionals considering together the appropriate timing to accomplish the family's wishes (P = 0.021). CONCLUSION: Many families have unfinished business. Health care professionals should coordinate the appropriate timing for what the family wishes to do, with consideration of family dynamics, including the family's preparedness, communication pattern, and relationships.

Quality of life of elderly patients on peritoneal dialysis versus hemodialysis: a single-center study.

BACKGROUND: The number of elderly dialysis patients in Japan is dramatically increasing. Receiving therapy with better satisfaction through home care is one of the important factors in their daily lives. Thus, the quality of life of elderly patients on hemodialysis (HD) or peritoneal dialysis (PD) was evaluated. METHODS: Clinical information of patients aged ≥80 years who started dialysis at our hospital between January 2013 and December 2015 was retrospectively collected. The mortality rate, length of hospitalization, and place of death were identified to evaluate patient quality of life. RESULTS: In total, 56 patients (14 PD and 42 HD) were enrolled. The mean age of study subjects was 85.2 ± 4.0 years. The proportion of PD patients who lived with their family or have professional caregivers who could assist them in their daily life was higher than that of HD patients (100 vs. 76.2%, respectively; p = 0.044). Mortality rate was higher in PD patients than in HD patients (p = 0.003), but long-term hospitalization of >180 days was observed only in HD patients (PD vs. HD: 0.0 vs. 16.7%; p = 0.102). In patients with Barthel index scores <100, the long-term hospitalization difference was significant (PD vs. HD: 0.0 vs. 30.4%; p = 0.040). Of note, 6 of 7 deceased PD patients and 1 of 10 deceased HD patients died at home (p = 0.002). CONCLUSION: PD is a desirable home care therapy for elderly patients, but the burden on caregivers should be considered.

Physiological and microbiological verification of the benefit of hair washing in patients with skin conditions of the scalp.

BACKGROUND: It is unclear whether hair washing is effective against scalp eruption and pruritus caused by seborrheic dermatitis or psoriasis vulgaris. AIMS: To assess whether a proper hair-washing regimen, including the use of antibacterial shampoo, can ameliorate scalp eruption symptoms and alter the composition of the scalp microflora. METHODS: Eighteen patients with seborrheic dermatitis or psoriasis vulgaris scalp eruptions were instructed in proper techniques of daily hair washing, rinsing, and shampooing, which they underwent for 12 weeks. They used control shampoo in weeks 1-4 and 9-12, and an antibacterial shampoo during weeks 5-8. At the start of the test period and at weeks 4, 8, and 12, we assessed scalp symptoms (erythema, scaling/desquamation, dryness, itchiness, and scratching scars); microbial DNA levels from lesion and nonlesion areas; and levels of interleukin (IL)-1α, IL-1ra, and total protein in the scalp's horny layer. RESULTS: Compared to baseline values, scaling/desquamation and itchiness improved significantly at weeks 8 and 12. Other observed skin symptoms also improved over time. Malassezia colonization levels in lesion and nonlesion areas decreased gradually; the decrease was significant at week 8 in lesion areas and at weeks 4 and 8 in nonlesion areas. Bacterial colonization levels also decreased gradually, achieving significance in lesion areas at week 4. Gradual decreases in IL-1ra/IL-1α level showed statistical significance at weeks 4 and 12, while the protein quantity significantly decreased at week 12. CONCLUSION: Proper hair washing improved scalp condition symptoms, and possibly the underlying etiology.

Characterization and cloning of GNA-like lectin from the mushroom Marasmius oreades.

A new mannose-recognizing lectin (MOL) was purified on an asialofetuin-column from fruiting bodies of Marasmius oreades grown in Japan. The lectin (MOA) from the fruiting bodies of the same fungi is well known to be a ribosome-inactivating type lectin that recognizes blood-group B sugar. However, in our preliminary investigation, MOA was not found in Japanese fruiting bodies of M. oreades, and instead, MOL was isolated. Gel filtration showed MOL is a homodimer noncovalently associated with two subunits of 13 kDa. The N-terminal sequence of MOL was blocked. The sequence of MOL was determined by cloning from cDNA and by protein sequencing of enzyme-digested peptides. The sequence shows mannose-binding motifs of bulb-type mannose-binding lectins from plants, and similarity to the sequences. Analyses of sugar-binding specificity by hemagglutination inhibition revealed the preference of MOL toward mannose and thyroglobulin, but asialofetuin was the strongest inhibitor of glycoproteins tested. Furthermore, glycan-array analysis showed that the specificity pattern of MOL was different from those of typical mannose-specific lectins. MOL preferred complex-type N-glycans rather than high-mannose N-glycans.

Variance analysis of a clinical pathway of video-assisted single lobectomy for lung cancer.

PURPOSE: Clinical pathways have contributed to standardized postoperative management, but analyzing variance is also important to maintain quality control. To evaluate the validity of our own clinical pathway for managing video-assisted lobectomy for lung cancer, we analyzed the variances influencing postoperative recovery. METHODS: Between April 2003 and April 2004, 62 consecutive patients with lung cancer underwent video-assisted single anatomic lobectomy with lymph node dissection. We evaluated 61 of these patients after the exclusion of one, who deviated from the clinical pathway management immediately as a result of serious complications. RESULTS: There were 29 men and 32 women, with an average age of 65.7 years. Complications developed in 29 patients, but there was no operative mortality. The occurrence of variance ranged from 1.6% to 34.4% for each activity and included prolonged supplemental oxygen therapy, out-of routine examination, prolonged epidural anesthesia, and delayed bathing frequently. Complications and an abnormal body mass index were significant and independent clinical factors affecting the increase in variance. CONCLUSIONS: Our original clinical pathway management was tolerable for lung cancer patients undergoing a video-assisted lobectomy. Complications and an abnormal body mass index were significant predictive factors for an increase in variance of our clinical pathway.

Rapid and sensitive diagnosis of adenoviral keratoconjunctivitis by loop-mediated isothermal amplification (LAMP) method.

PURPOSE: To develop a new method to detect and type adenoviruses directly from conjunctival scrapings using loop-mediated isothermal amplification (LAMP) with adenovirus (ad) type specific primer. METHODS: Using primers specific for the gene of ad1, ad3, ad4, ad8, ad19 and ad37, heat denatured adenovirus DNA was amplified by the LAMP and polymerase chain reaction (PCR). Alkaline lysed adenovirus prototype and conjunctival scrapings were also used directly as templates. RESULTS: Type specific primers amplified ad genes of the corresponding ad prototype specifically. The specific amplification was observed in both heat denatured and alkaline lysed samples. The amplified product was first detected within 45 min. Ad genotypes of clinical samples determined by the LAMP method were almost identical to those determined using the PCR-sequencing method. CONCLUSIONS: LAMP based isothermal amplification of adenovirus genome for detection and typing of adenoviruses is faster than PCR based methods. This new method will be useful for rapid diagnosis and typing of adenoviral conjunctivitis.

Rapid and sensitive diagnosis of adenoviral keratoconjunctivitis by loop-mediated isothermal amplification (LAMP) method.

PURPOSE: To develop a new method to detect and type adenoviruses directly from conjunctival scrapings using loop-mediated isothermal amplification (LAMP) with adenovirus (ad) type specific primer. METHODS: Using primers specific for the gene of ad1, ad3, ad4, ad8, ad19 and ad37, heat denatured adenovirus DNA was amplified by the LAMP and polymerase chain reaction (PCR). Alkaline lysed adenovirus prototype and conjunctival scrapings were also used directly as templates. RESULTS: Type specific primers amplified ad genes of the corresponding ad prototype specifically. The specific amplification was observed in both heat denatured and alkaline lysed samples. The amplified product was first detected within 45 min. Ad genotypes of clinical samples determined by the LAMP method were almost identical to those determined using the PCR-sequencing method. CONCLUSIONS: LAMP based isothermal amplification of adenovirus genome for detection and typing of adenoviruses is faster than PCR based methods. This new method will be useful for rapid diagnosis and typing of adenoviral conjunctivitis.

Digestion by serine proteases enhances salt tolerance of glutaminase in the marine bacterium Micrococcus luteus K-3.

Salt-tolerant glutaminase (Micrococcus glutaminase, with an apparent molecular mass of 48.3 kDa, intact glutaminase) from the marine bacterium Micrococcus luteus K-3 was digested using protease derived from M. luteus K-3. The digestion products were a large fragment (apparent molecular mass of 38.5 kDa, the glutaminase fragment) and small fragments (apparent molecular mass of 8 kDa). The digestion was inhibited by phenylmethanesulfonyl fluoride (PMSF). Digestion of intact glutaminase by serine proteases including trypsin, elastase, lysyl endopeptidase, and arginylendopeptidase also produced the glutaminase fragment. The N-terminus of the glutaminase fragment was the same as that of intact glutaminase. The N-termini of two small fragments were Ala394 and Ala396, respectively. The enzymological and kinetic properties of the glutaminase fragment were almost the same as those of intact glutaminase except for salt-tolerant behavior. The glutaminase fragment was a higher salt-tolerant enzyme than the intact glutaminase, suggesting that Micrococcus glutaminase is digested in the C-terminal region by serine protease from M. luteus K-3 to confer salt tolerance on glutaminase.

Heat treatment enhances healing process of experimental pseudomonas corneal ulcer.

We investigated the effects of hyperthermia on the healing process of experimental Pseudomonas corneal ulceration (PCU). Hartley guinea pigs were used to develop animal models of PCU. As a heat source, disposable chemical pocket warmers were applied. The healing process of PCU was compared between the heat-treated corneas and the control corneas. The severity of infection and the degree of angiogenesis were classified by a clinical scoring system. The animals were euthanized 14 days after infection and the corneas were submitted for histopathological examination. The expression of vascular endothelial growth factor (VEGF) was examined immunohistochemically. Comparative reverse transcription polymerase chain reaction was performed to measure the expression level of VEGF in the cornea. Hyperthermia significantly promoted corneal epithelization and neovascularization in the PCU model. Heat treatment significantly decreased the number of viable Pseudomonas organisms present in PCU. On immunohistochemistry, the heated cornea demonstrated more intense staining for VEGF. Comparative reverse transcription polymerase chain reaction showed upregulation of the expression level of VEGF mRNA in the heat-treated cornea. Hyperthermia accelerated the healing process of PCU with increased corneal neovascularization. Angiogenesis may play an important role in the PCU healing process, which is enhanced by the heat treatment.