Clinical significance of phenotypic features of blasts in patients with myelodysplastic syndrome

Knowledge of the blast phenotype in myelodysplastic syndrome (MDS) would be valuable, as in other malignancies, but remains sparse. This is mainly because MDS blasts are a minor population in clinical samples, making analysis difficult. Thus, for this blast phenotype study, we prepared blast-rich specimens (using a new density centrifugation reagent for harvesting blasts) from blood and marrow samples of 95 patients with various MDS subtypes and 21 patients with acute leukemia transformed from MDS (AL-MDS). Flow cytometry revealed that a high proportion of the enriched blast cells (EBCs) from almost all patients showed an immunophenotype of committed myeloid precursors (CD34(+)CD38(+)HLA-DR(+)CD13(+)CD33(+)), regardless of the disease subtype. The cytochemical reaction for myeloperoxidase was negative in 58% of the cases. Thus, the EBC phenotype is more immature in MDS than in de novo acute myeloid leukemia. MDS EBCs often coexpressed stem cell antigens and late-stage myeloid antigens asynchronously, but rarely expressed T- and B-lymphoid cell-specific antigens. Markers for myeloid cell maturation (CD10 and CD15) were more prevalent on EBCs from low-risk MDS (refractory anemia [RA] and RA with ringed sideroblasts), whereas markers for myeloid cell immaturity (CD7 and CD117) were more prevalent on EBCs from high-risk MDS (chronic myelomonocytic leukemia, RA with excess blasts [RAEB], and RAEB in transformation) and AL-MDS. A shift to a more immature phenotype of EBCs, accompanying disease progression, was also documented by sequential phenotyping of the same patients. Further, CD7 positivity of EBCs was an independent variable for a poor prognosis in MDS. These data represent new, valuable information regarding MDS.     

Tumor-associated macrophages and angiogenesis in early stage esophageal squamous cell carcinoma

Background

The association between angiogenesis and tumor-associated macrophages (TAMs) is unclear. Mononuclear cell infiltration was reported to induce angiogenesis in early stage esophageal squamous cell carcinoma (ESCC).

Methods

The study materials included 14 samples of normal squamous epithelium, 11 samples of low-grade intraepithelial neoplasia, and 64 samples of superficial esophageal cancer (M1 and M2 cancer 27; M3 or deeper cancer 37). We assessed microvessel density (MVD) using CD34 and CD105 immunostaining and monocyte count (MC) using CD68 and CD163 immunostaining in relation to the histological type or grade of mononuclear cell infiltration, as well as the correlation between MVD and MC.

Results

MVD and MC increased in accordance with histological type, and the differences were significant (P < 0.001). MVD and MC were significantly higher in M1 and M2 lesions than in normal squamous epithelium (P < 0.05). MVD (CD34 and CD105) and MC (CD68 and CD163) were significantly correlated with the degree of mononuclear cell infiltration (P < 0.001), and there was a strong correlation between MC assessed using CD68 and MC assessed using CD163 (rS = 0.93, P < 0.001). The CD163/CD68 ratio did not differ significantly according to histological type. There was a significant correlation between MVD assessed using CD105 and MC assessed using CD68 (rS = 0.69, P < 0.001) and CD163 (rS = 0.67, P < 0.001). MVD assessed using CD34 was also significantly correlated with MC assessed using both CD68 (rS = 0.59, P < 0.001) and CD163 (rS = 0.57, P < 0.001).

Conclusion

The number of TAMs is significantly associated with the development of neovasculature in the early stage of ESCC progression.

     

Change in tongue pressure in patients with head and neck cancer after surgical resection

Tongue pressure is reportedly associated with dysphagia. This study investigated relationships among characteristics of head and neck cancer, tongue pressure and dysphagia screening tests performed in patients with head and neck cancer during the acute phase after surgical resection. Fifty-seven patients (36 men, 21 women; age range 26–95 years) underwent surgical resection and dysphagia screening tests (Repetitive Saliva Swallowing Test, Water Swallowing Test, Modified Water Swallowing Test and Food Test) and pre- and postoperative measurement of tongue pressure at 5 time points (preoperatively, and 1–2 weeks and 1, 2, and 3 months postoperatively). Progression of cancer (stage), tracheotomy, surgical reconstruction, chemotherapy, radiotherapy and neck dissection were factors associated with postoperative tongue pressure. Data were analyzed by linear mixed-effect model, Spearman correlation coefficient and receiver operating characteristic (ROC) curve. Tongue pressure was significantly reduced 1–2 weeks after surgery, and recovered over time. Changes in tongue pressure were significantly associated with stage, radiotherapy and reconstruction. All screening tests showed a significant relationship with tongue pressure. Analysis of ROC and area under the effect curve suggested that a tongue pressure of 15 kPa can be used as a cut-off value to detect dysphagia after surgery for head and neck cancer. Our results suggest that tongue pressure evaluation might offer a safe, useful and objective tool to assess dysphagia immediately postoperatively in patients with head and neck cancer.     

Long Term Results and Predictors of Left Ventricular Function Recovery after Aortic Valve Replacement for Chronic Aortic Regurgitation

Objectives: In most patients with aortic regurgitation (AR), aortic valve replacement (AVR) improves left ventricular (LV) function, but some patients will not have favorable remodeling. Our objectives were to review long term clinical results of AVR for AR and to examine what factors affect the normalization of LV function after AVR for chronic AR.Methods: Between 1989 and 2010, 177 patients underwent isolated AVR for chronic pure AR. The patients were divided into 2 groups based on indexed end-systolic LV diameter (iESD): Group L (iESD) ≧25 mm/m²) (130 patients) and Group S (iESD <25 mm/m²) (47 patients).Results: There was no significant difference between groups in late mortality, freedom from cardiac-related death and rehospitalization for heart failure at late follow up after operation. At postoperative follow-up, 16% of patients had not recovered normal LV systolic function. By means of multivariate analysis, iESD and cardiac index (CI) were independent predictors of recovery of LV function and iESD >26.7 mm/m² and CI <2.71 l/min/m² were the best cut-off values.Conclusions: Early and late surgical results of AVR for chronic AR were good, but for the preservation of postoperative normal LV function, AVR for AR patients should be performed before iESD reaches 26.7 mm/m².     

Visceral Leishmaniasis Caused by Leishmania infantum in Salta,Argentina: Possible Reservoirs and Vectors

Cases of human visceral leishmaniasis (HVL) were not recorded until recently in the Chaco region of northwestern Argentina. Dogs were surveyed at the sites of infection of two HVL index cases in the Chaco region of Salta province. Canine cases (CanL) were diagnosed by two parasitological methods, two molecular methods targeting mini- and maxicircle DNA, and immunochromatographic dipstick. Among 77 dogs studied, 10 (13%) were found infected with Leishmania spp. In seven dogs and two humans, the infecting species was typed as Leishmania (Leishmania) infantum. The same genotype was detected in the human and two of the CanL. Although several diagnostic methods displayed weak or moderate agreement, the concordance values for serology versus maxicircle PCR were very good (Kappa index = 0.84). Sandflies captured in the area were identified as Lutzomyia migonei and Lu. cortelezzii/Lu. sallesi (cortelezzii complex). The focal appearance of leishmaniasis in dogs and humans in a sylvatic region and its relatively low prevalence of infection suggests that L. (L.) infantum transmission to dogs and humans may, in this region, stem from sylvatic reservoirs.     

Preliminary experience using a computer-mediated flexible circular stapler in laparoscopic esophagogastrostomy

Intracorporeal esophagogastrostomy after laparoscopic proximal gastrectomy is technically challenging. We employed a computer-mediated flexible circular stapler (SurgASSIST) for esophagogastrostomy in 2 cases with gastrointestinal stromal tumor located near the esophagogastric junction. Esophagogastrostomy was successfully constructed intracorporeally using the double-stapling technique. Operation times for the 2 cases were 225 and 170 minutes. No anastomotic leakage was encountered. However, anastomotic stricture requiring balloon dilatation occurred in 1 patient. The SurgASSIST system was feasible for esophageal anastomosis in laparoscopic proximal gastrectomy. However, the digital loading unit (DLU) is too large to introduce transorally, and attempting introduction of the DLU through the narrow lumen may create lesions or perforate the organ. Further improvements in the DLU will facilitate wider use of this system for various procedures in laparoscopic surgery.     

Study of interleukin-6 in the spread of colorectal cancer: the diagnostic significance of IL-6

We investigated the diagnostic significance of IL-6 for lymph node metastasis and/or hepatic metastasis from colorectal cancer in 65 patients and evaluated the contributions of 8 factors (IL-6, HGF, IL-1beta, TNF-alpha, TGF-beta1, ELAM-1, ICAM-1, VCAM-1) toward Dukes.s classification of 53 patients. We also examined IL-6 expression in tumor tissue. From the receiver operating characteristic (ROC) curve analysis, an optimal cutoff value of 5.8 pg/ml was determined to classify lymph node and/or hepatic metastasis, and that of 6.3 pg/ml was determined to classify hepatic metastasis. These values indicated sensitivities of 55.0% and 71.4%, and specifi cities of 100% and 88.6%, respectively. IL-6, HGF, and ELAM-1 were very useful for distinguishing among Dukes.s A/B group, C group, and D group. In all cases with high IL-6 values (more than 25.0 pg/ml), immunohistochemical staining was positive for IL-6 in the cytoplasm of cancer cells. IL-6 is strongly suspected to be involved in lymph node and/or hepatic metastasis by promoting it through HGF, and serum IL-6 value (pg/ml) would be useful diagnostically to estimate whether or not there is a high risk of lymph node and/or hepatic metastasis.     

Use of FTA Cards for Direct Sampling of Patients' Lesions in the Ecological Study of Cutaneous Leishmaniasis

The FTA card (Whatman) was assessed for its utility as a molecular epidemiological tool in collecting samples from patients with leishmaniasis in Peru because the card has a variety of merits; it is less invasive for patients and easy to handle for both physicians and other medical personnel for sample collection or diagnosis, in addition to its simplicity and easy countrywide and/or intercountry transportation for analysis. Samples were collected from 132 patients suspected of having leishmaniasis, and Leishmania species were successfully identified in samples from 81 patients in 15 departments of Peru by cytochrome b and mannose phosphate isomerase gene analyses. Of these, 61.7% were identified as Leishmania (Viannia) peruviana, 22.2% as L. (V.) braziliensis, 12.3% as L. (V.) guyanensis, 2.5% as L. (V.) shawi, and 1.2% as L. (V.) lainsoni. The three predominant species, L. (V.) peruviana, L. (V.) braziliensis, and L. (V.) guyanensis, were mainly found in the Andean highlands, in the tropical rainforest, and in northern and central rainforest regions, respectively. This is the first time L. (V.) shawi has been identified outside Brazil. The present study showed that the FTA card will be a useful tool for the ecological study of different forms of leishmaniasis. Furthermore, collecting samples directly from patients'' lesions by using the FTA card eliminates (i) the possibility of contamination of Leishmania isolates during short- and/or long-term passages of culture in vitro in each laboratory and (ii) pain and suffering of patients from taking samples by skin biopsy.Leishmaniasis is caused by protozoan parasites of the genus Leishmania, which is further divided into two subgenera, Leishmania (Leishmania) and Leishmania (Viannia) (10). The disease is widely distributed around the world, especially in tropical and subtropical areas, affecting at least 12 million people in 88 countries (6). Approximately 20 Leishmania species are known to be pathogenic to humans, and the species is the major determinant of clinical outcome (cutaneous, mucocutaneous, and visceral forms) (6). Therefore, identification of the parasite species in areas of endemicity is important for both appropriate treatment and prognosis.The standard method for the classification of Leishmania species is multilocus enzyme electrophoresis (MLEE), which requires the isolation and mass culture of the parasites (4, 17). This process has several disadvantages: (i) risk of contamination with bacteria and/or fungus and even other Leishmania isolates in the laboratory; (ii) maladaptation of the parasites to the artificial medium; (iii) difficulty in cultivation due to low numbers of parasites in cutaneous lesions, especially for the subgenus Leishmania (Viannia); and (iv) the time, several weeks or months, required to obtain a result after sample collection. All of these factors affect epidemiological studies in spite of considerable efforts to collect patient specimens from different areas of endemicity, especially for specimens from remote locations. To overcome these problems, molecular biological methods have been developed for the detection and identification of Leishmania species using DNA extracted from clinical samples (7, 16, 21). However, sampling procedures, such as skin biopsy, are sometimes painful for patients and become a burden to both patients and physicians. Therefore, alternative sampling procedures with less invasiveness, simple and easy handling, and greater convenience are required for the detection and identification of Leishmania species and continuous monitoring of endemic species of causative organisms.FTA technology (Whatman) is a rapid and safe method for extracting nucleic acids from blood, cell, and pathogen samples without using any organic solvent or specialized equipment. When the samples are spotted onto an FTA card, the cells are readily lysed and the nucleic acids are fixed on the card, resulting in protection from nuclease, oxidative, and UV damage and prevention of the growth of bacteria and other microorganisms. The card is also suitable for long-term storage and the transportation of materials at room temperature, eliminating the possibility of contamination from isolates in vitro during the laboratory phase. In the present study, the utility of FTA cards was assessed for sample collection for the countrywide molecular epidemiological study of leishmaniasis in Peru.     

Identification and validation of a gene expression signature that predicts outcome in malignant glioma patients

Better understanding of the underlying biology of malignant gliomas is critical for the development of early detection strategies and new therapeutics. This study aimed to define genes associated with survival. We investigated whether genes selected using random survival forests model could be used to define subgroups of gliomas objectively. RNAs from 50 non-treated gliomas were analyzed using the GeneChip Human Genome U133 Plus 2.0 Expression array. We identified 82 genes whose expression was strongly and consistently related to patient survival. For practical purposes, a 15-gene set was also selected. Both the complete 82 gene signature and the 15 gene set subgroup indicated their significant predictivity in the 3 out of 4 independent external dataset. Our method was effective for objectively classifying gliomas, and provided a more accurate predictor of prognosis. We assessed the relationship between gene expressions and survival time by using the random survival forests model and this performance was a better classifier compared to significance analysis of microarrays.