Bicarbonate in Arteries Measured Preoperatively for Cadaveric Single-lung Transplantation is Related to Intraoperative Extra-Corporeal Membrane Oxygenation Use: A Retrospective Preliminary Study.

Background: There are no known predictors of extracorporeal membrane oxygenation (ECMO) induction for single lung transplantation. Objective: The purpose of the present study was to clarify the relationship between variables and ECMO requirements in single lung transplantation. Methods: This study included adult patients who underwent cadaveric single lung transplantation between 2010 and 2019. After general anesthesia, the transplanted lungs were ventilated in all cases. The analysis included 38 patients in the ECMO required (RQ) group and 12 patients in the ECMO non-required (FR) group. Comparisons were made between the two groups for data affecting ECMO implementation, and data that were significantly different were subjected to multivariate analysis. Results: Prior to anesthesia, the bicarbonate (HCO3-) value of the FR group was lower than that of the RQ group (24.6±2.7 vs. 29.7±5.3 mmol/L, p=0.005). Multivariate analysis showed that the cut-off bicarbonate value was 29.6. The area under the receiver operating characteristic curve (AUROC) of the model was 0.869 (R2: 0.331), with a sensitivity of 79% and a specificity of 88%. The odds ratio was 1.63 for every unit increase in the bicarbonate value (95%CI: 1.11-2.39, p<0.001). Further, the FR group had higher arterial blood pressure (mean: 79.0±11.5 vs. 68.9±8.3 mmHg, p=0.030), less blood loss (432±385 vs. 1,623±1,997 g, p<0.001), shorter operation time (417±44 vs. 543±111 min, p<0.001), and shorter ICU stay (11±9 vs. 25±38 days, p=0.039). Conclusion: Preoperative evaluation of bicarbonate could predict the need for ECMO for single lung transplantation.

Sequence analysis of ABO and its homologues is valid for species identification.

BACKGROUND: ABO and its paralogues, such as A3GALT2 and GGTA1, encoding α1,3-Gal(NAc) transferases, belong to the glycosyltransferase 6 (GT6) gene family. We have developed an alternative method for the identification of species based on sequence variations within the GT6 gene family, which is applicable to degraded DNA. METHODS/MATERIALS: DNA samples prepared from control mammalian species, together with an unknown sample, were polymerase chain reaction (PCR)-amplified using one universal primer pair targeting the sequences in the last coding exons of the GT6 gene family, yielding 141-bp products derived from those multiple loci. After cloning, sequence determination and Basic Local Alignment Search Tool analysis, phylogenetic trees were constructed. RESULTS: Comparison of the sequences obtained with those references showed good concordance with each of the starting species of mammals. This system was able to identify 'mouse' or 'rodent' as the origin of the unknown sample. CONCLUSION: For the identification of species, genotyping of ABO and its homologues would be applicable for the analysis of degraded DNA samples. Although the method employed in this study is likely valid for mammals, it would not be suitable for birds, fish and reptiles. It may be possible to improve the present method for use with other species by employing an alternative universal primer set.

ABO alleles are linked with haplotypes of an erythroid cell-specific regulatory element in intron 1 with a few exceptions attributable to genetic recombination.

Recent investigation of transcriptional regulation of the ABO genes has identified a candidate erythroid cell-specific regulatory element, named the +5·8-kb site, in the first intron of ABO. Six haplotypes of the site have been reported previously. The present genetic population study demonstrated that each haplotype was mostly linked with specific ABO alleles with a few exceptions, possibly as a result of hybrid formation between common ABO alleles. Thus, investigation of these haplotypes could provide a clue to further elucidation of ABO alleles.

Blood group B gene is barely expressed in in vitro erythroid culture of Bm-derived CD34+ cells without an erythroid cell-specific regulatory element.

BACKGROUND AND OBJECTIVES: Previously, a weak phenotype Am or Bm was assumed to be caused by a reduction of A or B gene expression in bone marrow cells, but not in mucus-secreting cells. However, ABO expression has not been examined in erythroid progenitor cells of Am or Bm individuals. MATERIALS AND METHODS: We carried out in vitro erythroid differentiation of CD34(+) cells from peripheral blood of a Bm individual harbouring a 3.0-kb deletion including an erythroid cell-specific regulatory element, named the +5.8-kb site, in intron 1 of the human ABO blood group gene. RESULTS: During the in vitro differentiation of CD34(+) cells from this Bm individual into erythroid cells, B-antigens were not detectable on the cultured cells by flow cytometric analysis, and allele-specific RT-PCR consistently detected the transcripts from the O allele, but not from the B allele. Moreover, chromatin immunoprecipitation assay demonstrated that both RUNX1 and GATA-2 or GATA-1 were bound to the +5.8-kb site in cultured erythroid cells expressing ABO. CONCLUSION: It is likely that the +5.8-kb site enhances transcription from the ABO promoter in erythroid cells through binding of RUNX1 and GATA-2 or GATA-1.

A 3.0-kb deletion including an erythroid cell-specific regulatory element in intron 1 of the ABO blood group gene in an individual with the Bm phenotype.

We developed a sequence-specific primer PCR (SSP-PCR) for detection of a 5.8-kb deletion (B(m) 5.8) involving an erythroid cell-specific regulatory element in intron 1 of the ABO blood group gene. Using this SSP-PCR, we performed genetic analysis of 382 individuals with Bm or ABm. The 5.8-kb deletion was found in 380 individuals, and disruption of the GATA motif in the regulatory element was found in one individual. Furthermore, a novel 3.0-kb deletion involving the element (B(m) 3.0) was demonstrated in the remaining individual. Comparisons of single-nucleotide polymorphisms and microsatellites in intron 1 between B(m) 5.8 and B(m) 3.0 suggested that these deletions occurred independently.

Presence of nucleotide substitutions in transcriptional regulatory elements such as the erythroid cell-specific enhancer-like element and the ABO promoter in individuals with phenotypes A3 and B3, respectively.

BACKGROUND AND OBJECTIVES: An erythroid cell-specific regulatory element, referred to as the +5.8-kb site, has been identified in the first intron of the human ABO blood group gene. Subsequent studies have revealed involvement of deletion or mutation at the site in phenotypes Am , Bm and ABm . We investigated the molecular mechanisms involved in the A3 and B3 phenotypes. MATERIALS AND METHODS: Genomic DNAs were prepared from peripheral blood of seven A3 individuals and twelve B3 or AB3 individuals, and the nucleotide sequences were investigated using PCR and sequencing. Promoter assays were performed with K562 cells. RESULTS: Two single point-mutations at +5893 or +5909 in the site on the A-allele were found in A3 individuals, while promoter assays revealed decreased activity at the site as a result of each substitution. In two B3 individuals, a single point-mutation at -77 in the ABO promoter on the B-allele was found, and the substitution was demonstrated to reduce the promoter activity. CONCLUSION: Nucleotide substitutions in the transcriptional regulatory elements such as the +5.8-kb site and the ABO promoter appear to decrease transcription from the A- and B-alleles, resulting in reduction in A- and B-antigen expression in A3 and B3, respectively.

Deletion of the RUNX1 binding site in the erythroid cell-specific regulatory element of the ABO gene in two individuals with the Am phenotype.

BACKGROUND AND OBJECTIVES: An erythroid cell-specific regulatory element, referred to as the +5·8-kb site, had been identified in the first intron of the human ABO blood group gene. Subsequent studies revealed that either a 5·8-kb deletion including the +5·8-kb site or disruption of a GATA factor binding motif at the site was present in all Bm and ABm individuals examined. We investigated the molecular mechanism of the Am phenotype, which is analogous to the Bm phenotype. MATERIALS AND METHODS: Genomic DNAs were prepared from peripheral blood of two Am individuals, and the nucleotide sequences were investigated using PCR and direct sequencing. Electrophoretic mobility shift assay (EMSA) and promoter assay with K562 cells were carried out. RESULTS: A novel 23-bp nucleotide deletion was found at the +5·8-kb site in both individuals. EMSAs demonstrated binding of the transcription factor RUNX1 to the nucleotides within the deletion. Promoter assays showed that the deletion reduced the transcriptional activity of the +5·8-kb site. CONCLUSION: Deletion of the 23-bp nucleotides including the RUNX1 binding site decreases transcription of the A allele, resulting in the reduction in A antigen expression in the Am phenotype.

Analysis of cell cycle-related proteins in mediastinal lymph nodes of patients with N2-NSCLC obtained by EBUS-TBNA: relevance to chemotherapy response.

BACKGROUND: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is an accurate tool for lymph node staging of non-small cell lung cancer (NSCLC). Most patients with NSCLC require systemic chemotherapy during their treatment, with relatively poor responses. If the response to chemotherapy could be predicted, ideally at the time of the initial bronchoscopic examination, the therapeutic benefit could be maximised while limiting toxicity. A study was therefore undertaken to investigate the feasibility of EBUS-TBNA for obtaining tissue samples from mediastinal lymph nodes that can be used for immunohistochemical analysis, and to stratify patients with molecular-based pN2-NSCLC into chemo-responsive and chemoresistant subgroups who might benefit from tailoring of chemotherapy. METHODS: The expression of six cell cycle-related proteins (pRb, cyclin D1, p16(INK4A), p53, p21(Waf1), Ki-67) in mediastinal lymph node specimens obtained by EBUS-TBNA was investigated by immunohistochemistry in 36 patients with pN2-NSCLC. Their predictive role(s) in the response to platinum-based chemotherapy was examined. RESULTS: Immunostaining was feasible in all studied specimens. Univariate analysis revealed that p53 and p21(Waf1) expressions were significantly related to the response to chemotherapy (p = 0.002 and p = 0.011, respectively). Multivariate logistic regression analysis revealed that only p53 overexpression was associated with a poor response to chemotherapy (p = 0.021). CONCLUSIONS: These results suggest that EBUS-TBNA is a feasible tool for obtaining mediastinal nodal tissue samples amenable for immunohistochemical analysis. Immunostaining of p53 in EBUS-TBNA-guided specimens may be useful in predicting the response to chemotherapy in patients with N2-NSCLC and helping in the selection of patients who might benefit from certain chemotherapeutic strategies.

Evaluation of a novel kit for the rapid detection of extended-spectrum beta-lactamases.

Since a method of rapidly detecting extended-spectrum beta-lactamase (ESBL) production in gram-negative isolates from patients with severe infection is urgently required, the present study of a novel commercial kit was conducted. The Cica-Beta Test I (Kanto Chemical, Tokyo, Japan) is designed for the rapid detection of ESBL in gram-negative bacteria directly from isolated colonies in a 15-min protocol. In this study, a total of 304 strains of Klebsiella spp., Escherichia coli and Proteus mirabilis were tested using the novel kit and the phenotypic confirmatory disk test using cefotaxime and ceftazidime with and without clavulanate. The kit showed 95.5 and 98.1% sensitivity and specificity, respectively, as compared to the disk test, and thus proved to be an appropriate tool for the rapid detection of ESBL.

Metabolism of 2-nitrofluorene, 2-aminofluorene and 2-acylaminofluorenes in rat and dog and the role of intestinal bacteria.

1. The in vivo metabolism of 2-nitrofluorene (NF), an environmental pollutant, and 2-aminofluorene (AF) and its acylated derivatives, 2-formylaminofluorene (FAF) and 2-acetylaminofluorene (AAF), was examined in rat and dog. 2. 7-Hydroxy-2-nitrofluorene, 5-hydroxy-2-nitrofluorene, AF, AAF, FAF, 7-hydroxy-2-aminofluorene, 5-hydroxy-2-aminofluorene, 7-hydroxy-2-acetylaminofluorene, 5-hydroxy-2-acetylaminofluorene, 7-hydroxy-2-formylaminofluorene and 5-hydroxy-2-formylaminofluorene were identified as urinary and faecal metabolites of NF in rat and dog. 3. AAF and its hydroxylated derivatives were detected as major metabolites of NF in rat, but FAF and its hydroxylated metabolites were mainly excreted in dog. 4. AF, AAF, FAF and their hydroxylated metabolites were also identified as urinary and faecal metabolites of AF, AAF or FAF in rat, suggesting that AAF and FAF are interconverted via AF. 5. Treatment of rat and dog with antibiotics significantly decreased the urinary and faecal excretion of AF and its derivatives after oral administration of NF, and partly decreased the excretion of acylated metabolites after an oral dose of AF. 6. The caecal contents of untreated rats and some species of intestinal bacteria exhibited nitro-reductase activity toward NF, and acylating activity toward AF, affording AAF and FAF.

Effects of consonantal context on perceptual assimilation of American English vowels by Japanese listeners.

This study investigated the extent to which adult Japanese listeners' perceived phonetic similarity of American English (AE) and Japanese (J) vowels varied with consonantal context. Four AE speakers produced multiple instances of the 11 AE vowels in six syllabic contexts /b-b, b-p, d-d, d-t, g-g, g-k/ embedded in a short carrier sentence. Twenty-four native speakers of Japanese were asked to categorize each vowel utterance as most similar to one of 18 Japanese categories [five one-mora vowels, five two-mora vowels, plus/ei, ou/ and one-mora and two-mora vowels in palatalized consonant CV syllables, C(j)a(a), C(j)u(u), C(j)o(o)]. They then rated the "category goodness" of the AE vowel to the selected Japanese category on a seven-point scale. None of the 11 AE vowels was assimilated unanimously to a single J response category in all context/speaker conditions; consistency in selecting a single response category ranged from 77% for /eI/ to only 32% for /ae/. Median ratings of category goodness for modal response categories were somewhat restricted overall, ranging from 5 to 3. Results indicated that temporal assimilation patterns (judged similarity to one-mora versus two-mora Japanese categories) differed as a function of the voicing of the final consonant, especially for the AE vowels, /see text/. Patterns of spectral assimilation (judged similarity to the five J vowel qualities) of /see text/ also varied systematically with consonantal context and speakers. On the basis of these results, it was predicted that relative difficulty in the identification and discrimination of AE vowels by Japanese speakers would vary significantly as a function of the contexts in which they were produced and presented.

The masking effect in foreign speech sounds perception revealed by neuromagnetic responses.

The backward masking effect on non-native consonants by a following vowel was examined using neuromagnetic responses to synthesized speech sounds. Native speakers of Japanese were presented with sequences of frequent (85%) and infrequent (15%) speech sounds (/ra/ and /la/ respectively, no /l/ /r/ contrast in Japanese language). The duration of the stimuli was 110 ms in a short session and 150 ms in a long session. In the short session, the stimuli were terminated in the course of the transition from the consonant to the vowel to diminish the masking effect from the vowel part. A distinct magnetic counterpart of mismatch negativity (MMNm) was observed for the short session, whereas a smaller MMNm was observed for the long session.

Horizontal inclination of the longitudinal axis of the colonic J-pouch: defining causes of evacuation difficulty.

PURPOSE: Functional outcome after low anterior resection for rectal cancer is improved by the construction of a colonic J-pouch. One disadvantage of this type of reconstruction is evacuation difficulty, which has been associated with large pouches. The purpose of this study was to elucidate the causes of evacuation difficulty in large pouches using pouchography. METHODS: The angle between the longitudinal axis of the pouch and the horizontal line (pouch-horizontal angle) on lateral pouchography was determined in 26 patients with 10-cm J-pouch reconstructions (10-J group) and 27 patients with 5-cm J-pouch reconstructions (5-J group). Measurement were made at three months, one year, and two years after surgery. Clinical function was evaluated using a questionnaire one year postoperatively. RESULTS: The pouch-horizontal angle in the 10-J group was significantly smaller than that in the 5-J group at all three time points. In both groups the pouch-horizontal angle at one year was significantly smaller than that at three months. There were no significant differences between the pouch-horizontal angles at one and two years. An evacuation difficulty was significantly more common in the 10-J group than the 5-J group. CONCLUSIONS: The evacuation difficulty observed in patients with large colonic J-pouch reconstructions may be attributed to the development of a horizontal inclination within one year of surgery.

High ligation of the inferior mesenteric artery with hypogastric nerve preservation in rectal cancer surgery.

We describe herein a technique of performing upward node dissection following high ligation of the inferior mesenteric artery for patients with T3 and T4 rectal carcinomas. The course of the hypogastric nerve is confirmed macroscopically during the procedure to ensure its preservation. This technique offers both increased radicality and the prevention of ejaculatory dysfunction.

Examination of nodal metastases by a clearing method supports pelvic plexus preservation in rectal cancer surgery.

PURPOSE: In rectal cancer surgery preservation of urinary and sexual function is attempted by means of operations preserving the autonomic nerves of the pelvic plexus. Emergence of residual cancer because of a more shallow plane of dissection is a problem of concern with these methods, so we examined indications for pelvic plexus preservation. METHODS: We studied 198 patients with rectal carcinoma who underwent abdominopelvic lymphadenectomy. Lymph nodes along the superior hemorrhoidal artery and middle hemorrhoidal artery medial to the pelvic plexus were defined as perirectal nodes, and nodes along the middle hemorrhoidal artery lateral to the pelvic plexus and along the internal iliac artery represented lateral intermediate nodes. Node metastases were examined by the clearing method. RESULTS: Metastasis to perirectal nodes occurred in 12.5 percent in patients with pT1 tumors, 28.9 percent of those with pT2 tumors, and 50.0 percent of those with rectosigmoid junctional cancer. Metastasis to lateral intermediate nodes was absent in patients with pT1 or pT2 tumors and was as low as 2.5 percent in patients with rectosigmoid junctional cancer. CONCLUSIONS: In patients with T1, T2, and rectosigmoid junctional cancer, perirectal node dissection is necessary, but chances of residual cancer should remain minimal when the pelvic plexus is preserved.

Lipopeptide particles as the immunologically active component of CTL inducing vaccines.

Using a bipalmitoylated lipopeptide consisting of an ovalbumin helper T-cell epitope covalently linked to an influenza virus cytotoxic T-lymphocyte (CTL) epitope, we addressed possible factors that may be critical for CTL induction. Antigen processing of lipopeptide appears to be required for T-cell induction since there was virtually no in vitro binding of lipopeptide to purified MHC molecules. A major portion of lipopeptide immunogenicity was due to its particulate nature inasmuch as CTL induction in mice correlated with insoluble lipopeptide constructs, whereas more soluble analogs were significantly less immunogenic. Immunohistological analysis of tissue from immunized animals revealed that lipopeptide migration from the s.c. injection site to the spleen could be detected as early as 1 h after immunization and cell-associated lipopeptide was observed on macrophages and dendritic cells, implicating both cell populations in the processing and presentation of lipopeptide particles to CTLs.

[Ocular blood flow defect in gaze-evoked amaurosis].

PURPOSE: To investigate the mechanism of gaze-evoked amaurosis. CASE: A patient with an optic nerve tumor presented gaze-evoked amaurosis. A large central scotoma and constricted visual field were detected after shifting the gaze laterally. However, the patient did not notice this change until his fellow eye was occluded. Computed tomography/magnetic resonance imaging (CT/MRI) demonstrated a tubularly enlarged optic nerve extending for 1 cm from the eye ball, and the condition was judged to be optic nerve sheath meningioma. He underwent fluorescein fundus angiography twice, once during straight gaze and again during lateral gaze. RESULTS: Manifestly decreased blood flow in the central retinal artery and filling defect in the optic nerve head were seen during lateral gaze. CONCLUSION: Forced lateral gaze may have increased the intra-optic nerve pressure at the optic nerve head and decreased the blood flow of the central retinal artery and peripapillary choroidal flow irrigating the optic nerve head.

Coloanal anastomosis using a circular stapling device following perineal rectosigmoidectomy for rectal prolapse.

Perineal rectosigmoidectomy with a hand-sewn anastomosis is thought to be the most appropriate procedure for elderly patients deemed unfit to tolerate a major abdominal operation. However, the use of a circular stapling device to perform the coloanal anastomosis following rectosigmoidectomy shortens the operative time and provides a more secure anastomosis than the traditional hand-sewn technique.

An evaluation of neural invasion in esophageal cancer.

It is well known that the operative results for esophageal cancer, especially thoracic esophageal cancer, are not favorable. We analyzed the relationship between neural invasion (NI) and histopathologic factors and recurrence types in 104 patients who underwent resection of esophageal cancers with T2 or greater depth of invasion of the esophageal wall. The implications of NI as a prognostic indicator were also examined. Of the 104 patients, 48 (46.2%) were NI-positive (NI(+)) and 56 (53.8%) were NI-negative (NI(-)). The NI(+) patients had a higher ratio of type 3 cancer. Concerning the histopathologic factors, there was a significant relationship between NI and lymph node metastasis (N) and between NI and lymphatic vessel invasion (ly) (P < 0.05). Examining the types of recurrence, namely hematogenous, lymphogenous, and local/stump, as well as pleural or peritoneal dissemination, a relationship was observed between lymphogenous recurrence and N or ly, and between local/stump recurrence and NI. The prognosis of the NI(+) patients was significantly different from that of the NI(-) patients. According to a multivariate analysis, NI and N were significant prognostic factors. These findings demonstrate that NI is an important prognostic factor closely related to local recurrence in patients with esophageal cancer. Thus, when treating advanced esophageal cancer with T2 or greater depth of invasion, NI and lymph node excision should be considered.

Indication for using high ligation of the inferior mesenteric artery in rectal cancer surgery. Examination of nodal metastases by the clearing method.

PURPOSE: In surgery for rectal cancer, it is unclear whether the inferior mesenteric artery should be ligated at a high or low position. The study contained herein was undertaken to clarify the indications for high ligation of the inferior mesenteric artery. METHODS: Subjects included 198 patients with rectal cancer who underwent resection with high ligation of the inferior mesenteric artery. Nodal metastases were examined by the clearing methods. RESULTS: The incidence of metastases to the lymph nodes surrounding the origin of the inferior mesenteric artery (root nodes) was 8.6 percent. Inferior mesenteric artery root nodal metastases occurred more frequently with pT3 and pT4 cancer. The five-year survival rate in patients with inferior mesenteric artery root nodal metastases was 38.5 percent; this rate was significantly lower than in those without inferior mesenteric artery root nodal metastases (73.4 percent). CONCLUSIONS: Although the five-year survival rate in patients with inferior mesenteric artery root nodal metastases was lower than in those without metastases, inferior mesenteric artery root nodal dissection should be performed after high ligation of the inferior mesenteric artery for patients with pT3 and pT4 cancers.