Prevalence of the genus Cladosporium on the integument of leaf-cutting ants characterized by 454 pyrosequencing.

The relationship of attine ants with their mutualistic fungus and other microorganisms has been studied during the last two centuries. However, previous studies about the diversity of fungi in the ants' microenvironment are based mostly on culture-dependent approaches, lacking a broad characterization of the fungal ant-associated community. Here, we analysed the fungal diversity found on the integument of Atta capiguara and Atta laevigata alate ants using 454 pyrosequencing. We obtained 35,453 ITS reads grouped into 99 molecular operational taxonomic units (MOTUs). Data analysis revealed that A. capiguara drones had the highest diversity of MOTUs. Besides the occurrence of several uncultured fungi, the mycobiota analysis revealed that the most abundant taxa were the Cladosporium-complex, Cryptococcus laurentii and Epicoccum sp. Taxa in the genus Cladosporium were predominant in all samples, comprising 67.9 % of all reads. The remarkable presence of the genus Cladosporium on the integument of leaf-cutting ants alates from distinct ant species suggests that this fungus is favored in this microenvironment.

Predicting initial recurrence pattern of esophageal cancer after neoadjuvant chemotherapy.

BACKGROUND/AIMS: No report has reviewed which clinicopathological factors including 3-field dissection and the response to neoadjuvant chemotherapy can predict the recurrence pattern of an esophageal carcinoma. The aim of this study was to reveal clinicopathological predictors for the initial recurrence pattern of a thoracic esophageal carcinoma. METHODOLOGY: Sixteen parameters derived from 98 patients who underwent a curative esophagectomy with neoadjuvant chemotherapy for a squamous cell carcinoma of the thoracic esophagus were examined using univariate and multivariate logistic regression analyses. RESULTS: Thirty-seven (37.8%) of the 98 patients had recurrences (hematogenous; 16, lymphatic; 13, others; 8). Univariate analyses revealed that the completion of 3-field dissection was the only factor for suppressing the lymphatic recurrence (P = 0.009; odds ratio: 0.2). Multivariate analyses showed that the number of positive nodes was a significant predictor for recurrence including all modalities (P = 0.02; odds ratio: 1.2) and both the number of positive nodes (P = 0.04; odds ratio: 1.1) and the poor response to neoadjuvant chemotherapy (P = 0.02; odds ratio: 6.9) were significant predictors for the hematogenous recurrence. CONCLUSIONS: The number of positive nodes and the response to neoadjuvant chemotherapy could predict the hematogenous recurrence of esophageal carcinoma.

Accelerated growth signals and low tumor-infiltrating lymphocyte levels predict poor outcome in T4 esophageal squamous cell carcinoma.

BACKGROUND: Little is known about the biological nature of T4 esophageal carcinoma growth signals and host defenses. METHODS: Paraffin-embedded sections from 78 patients with T2 to T4 esophageal squamous cell carcinoma who underwent operation were analyzed with immunohistochemistry. RESULTS: Positive cyclin A showed a significantly greater increase in T4 tumors than in those of other stages, and negative p27 showed a significantly greater decrease in T4 tumors than in large T3 stage tumors (tumor size > or = 4.0 cm). Patients with low-grade tumor-infiltrating lymphocyte (TIL) density showed a significantly greater decrease in T4 than in T2. The combination of p27 and cyclin A was a significant independent prognostic factor among T and N factors in multivariate analysis. TIL density was an independent prognostic factor among immunonutritional variables such as serum albumin concentration and the number of total blood lymphocytes. CONCLUSIONS: T4 esophageal squamous cell carcinoma has a poor prognosis, which is associated with increased p27-negative and cyclin A-positive growth signals in the tumor and with low TIL density in the host.

[Vascular anastomoses in free jejunal reconstruction to the neck vessels: an experience of consecutive 20 cases without using surgical microscope].

Twenty consecutive cases of pharyngoesophageal cancer who underwent free jejunal reconstruction were reported. The common carotid or external carotid artery was used for a feeder of the free graft. The internal jugular vein were served as a drainage vein. All anastomoses were performed in an end-to-side fashion without using surgical microscopes. Mean carotid artery clamping time was 16 minutes and no neurological complications were noticed postoperatively. Graft failure was occurred in 1 patient. The presenting technique, showing 95% success rate, is recommended as a simple option for vascular anastomosis in free jejunal reconstructive surgery.

Recurrent nerve nodal involvement is associated with cervical nodal metastasis in thoracic esophageal carcinoma.

BACKGROUND: Because three-field dissection for esophageal carcinoma is one of the most invasive operations, this procedure should be selected only when strictly indicated; but there are no useful criteria for it. The goal of this study was to identify the useful clinicopathologic factors for indicating three-field dissection. STUDY DESIGN: In this study, we reviewed the survival of patients after three-field dissection and identified factors associated with metastases in cervical nodes (CN), especially internal jugular nodes and supraclavicular nodes. Eighty-six patients who underwent curative esophagectomy with three-field dissection for squamous cell carcinoma of the thoracic esophagus were enrolled in this study. Survival rates were compared with respect to the presence of nodal metastasis. The relationship between recurrent nerve nodal (RNN) involvement and CN metastasis (bilateral internal jugular nodes, supraclavicular nodes, or both) was examined. Clinicopathologic factors possibly influencing CN metastasis were studied by multivariate logistic regression analysis. RESULTS: The overall 5-year survival rate was 45.1%. The 5-year survival rate for patients without metastatic nodes was 67.5%, for patients with one to four metastatic nodes it was 53.1%, and for patients with five or more it was 9.1 %. The prognosis of those with five or more metastatic nodes was significantly poorer than those of the other two groups. In the positive-node group, the 5-year survival rate for patients with RNN metastasis was 21.7%, and for patients with negative RNN it was 47.0% (p = 0.2). In the positive-node group, the 5-year survival rate for patients with positive CN was 13.7% and for patients with negative CN it was 45.8% (p = 0.01). Fifty-six (88.9%) of 63 patients without RNN metastasis had no CN metastasis in contrast to 13 of 23 patients (56.5%) with RNN metastasis who had no CN metastasis (p = 0.001). The positive predictive value, negative predictive value, sensitivity, and specificity were 43.5%, 88.8%, 58.8%, and 81.2%, respectively. The number of metastatic nodes (5 or more versus 0-4) (odds ratio: 2.9, 95% Confidence Interval (CI) = 1.6-5.5, p = 0.0008) and RNN involvement (odds ratio: 4.5, 95% CI = 1.3-15.9, p = 0.02) were the significant factors associated with CN metastasis in the multivariate analysis. CONCLUSIONS: RNN involvement is associated with CN metastasis as is the number of metastatic nodes and may be an indicator for the selection of three-field dissection in thoracic esophageal carcinoma.

Is upper mediastinal lymphadenectomy necessary in squamous carcinoma of the lower thoracic oesophagus?

AIM: We examined the indication of upper mediastinal lymphadenectomy for a squamous cell carcinoma of the lower thoracic oesophagus. METHODS: 49 patients underwent a curative oesophagectomy with upper mediastinal lymphadenectomy for a squamous cell carcinoma of the lower thoracic oesophagus. Node status and clinicopathological characteristics of these patients were reviewed retrospectively. RESULTS: 16 (94.1%) of 17 patients with superficial tumours had no positive node in the upper mediastinum. Nine (29.0%) of 31 patients with transmural tumours had positive nodes in the upper mediastinum (P = 0.04). Ten (20.4%) of 49 patients had many positive nodes in the upper mediastinum. Of these 10 patients, 6 patients had 5 or more positive nodes in all. The 5-year survival rate for patients with 5 or more positive nodes was 7.7%, which was significantly poorer than patients with 4 or fewer positive nodes. CONCLUSIONS: Upper mediastinal lymphadenectomy is unnecessary in most of the superficial squamous carcinomas of the lower thoracic oesophagus.

[A case of advanced gastric cancer successfully treated by neoadjuvant chemotherapy with CPT-11, CDDP and 5-FU].

A patient with advanced gastric cancer was treated with combined administration of CPT-11 CDDP and 5-FU before operation. CPT-11 was given intravenously at a dose of 30 mg/m2/day on day 1 and day 8. At the same time, 5 mg/m2/day CDDP and 350 mg/m2/day 5-FU were infused for 2 weeks. The patient experienced no other adverse reaction than a mild degree of nausea. Histological examination of the resected specimen revealed complete disappearance of cancer cells both in the stomach and the regional lymphnodes.

Stabilometric assessment in the anterior cruciate ligament-reconstructed knee.

OBJECTIVE: To examine the hypotheses that anterior cruciate ligament (ACL) reconstruction improves the proprioception of the knee beyond the level of ACL-deficient knees, and that proprioception of the knee correlates well with knee function after ACL reconstruction. PATIENTS: Fifty-three patients with ACL-reconstructed knees (22 men and 31 women), 30 physically active healthy volunteers with normal knees (15 men and 15 women), and 30 patients with chronic ACL-deficient knees (15 men and 15 women). MAIN OUTCOME MEASURES: A stabilometric assessment, which is considered to be a useful method for evaluating proprioception objectively, was used to compare the one-leg standing balance among three groups; and the relationship between subjective knee function, satisfaction, hop index, knee laxity, isokinetic thigh muscle strength, and one-leg standing balance in the ACL-reconstructed patients was also analyzed. RESULTS: The one-leg standing balance of the patients with ACL-reconstructed knees was still impaired compared with that of healthy volunteers (men, p < 0.05; women, p < 0.01), but significantly better than that of the patients with ACL-deficient knees (men, p < 0.001; women, p < 0.001). The one-leg standing balance of the patients with ACL-reconstructed knees correlated well with their functional outcomes such as subjective knee function (men, p < 0.001; women, p < 0.01), satisfaction (men, p < 0.01; women, p < 0.001), and hop index (men, p < 0.001; women, p < 0.001), whereas poor correlation was seen between functional outcomes and mechanical stability. CONCLUSIONS: The results suggest that knee function after ACL-reconstruction should be closely related with knee proprioception, and indicate that the usefulness of stabilometric assessment in the evaluation of the function of ACL-reconstructed knees.

The conservative treatment of complete tears of the anterior cruciate ligament in skeletally immature patients.

We describe the results of conservative treatment for complete midsubstance tears of the anterior cruciate ligament (ACL) in 18 skeletally immature patients, followed for a minimum of 36 months. Six patients had an ACL reconstruction during the follow-up period and were assessed immediately before their operation. The average time from initial injury to evaluation was 51 months. All patients had symptoms when reviewed. The modified Lysholm knee score showed one excellent result, one good, eight fair, and eight poor with a mean score of 64.3. Only one patient had returned to her preinjury level of athletics. Secondary meniscal tears were confirmed in six patients, and three more had the clinical signs of a tear at follow-up. Radiological evidence of degenerative changes was found in 11 of the 18 patients. We conclude that the results of non-operative treatment for ACL injuries in this age group are poor and not acceptable.

[Assessment of nucleolar organizer regions (Ag-NORs) in bronchial dysplasia and roentgenographically occult bronchogenic squamous cell carcinoma].

Mean argyrophil nucleolar organizer regions (Ag-NORs) per nucleus were determined on columnar epithelium, bronchial dysplasia, roentgenographically occult squamous cell carcinoma and squamous cell carcinoma with roentgenographically findings in 76 formalin-fixed, paraffin-embedded resected specimens. 1) Mean Ag-NOR counts per nucleus (mean +/- standard deviation) were 1.67 +/- 0.30 in 10 columnar epitheliums, 2.74 +/- 0.41 in 13 lesions with bronchial dysplasia, 3.75 +/- 0.59 in 57 lesions with roentgenographically occult squamous cell carcinoma and 4.44 +/- 0.51 in 14 lesions with squamous cell carcinoma with roentgenographically findings. There were significant differences between each two groups. 2) In lesions with squamous call carcinoma, mean Ag-NOR counts per nucleus of T2, T3 or T4 tumor were significantly higher than those of T1 tumor. Mean Ag-NOR counts per nucleus of N1 or N2 tumor were significantly higher than those of N0 tumor.

Multicentricity in resected occult bronchogenic squamous cell carcinoma.

The frequency and the treatment of multicentricity in 127 patients with resected roentgenographically occult bronchogenic squamous cell carcinoma were studied. The cumulative rate and the incidence of postoperative metachronous multiple primary lung cancer were 0.11 at 5 years after initial operation and 0.022 per patient-year, respectively. The cumulative rate and the incidence of second primary lung cancer, which includes synchronous and subsequent metachronous cancer in patients with initial lung cancer, were 0.17 at 5 years after the initial operation and 0.041 per patient-year, respectively. The cumulative rate and the incidence of third primary lung cancer in patients with second primary lung cancer were 0.47 at 5 years, which was significantly higher (p = 0.05) than that of second primary lung cancer, and 0.11 per patient-year, respectively. In all 12 patients with synchronous multiple primary lung cancer, no recurrence was observed after treatment, but 3 had subsequent multiple primary lung cancer. Among the 13 patients with postoperative metachronous multiple primary lung cancer, recurrence was observed in 1 of the 6 patients who underwent resection and in 2 of the 4 patients treated with laser or radiation therapy or both. The overall survival rate at 5 years after initial operation in patients with solitary and those with multicentric occult bronchogenic squamous cell carcinomas was 0.90 and 0.59, respectively.

Localization of double, roentgenographically occult lung cancer. Cytologic findings from selective brushing of all segmental and subsegmental bronchi.

Using selective brushing of all segmental and subsegmental bronchi, six patients were diagnosed as having synchronous, double, roentgenographically occult lung cancers. Experienced bronchoscopists failed to detect four "second cancer" lesions in six patients. The appearance of atypical cells as shown by cytologic examination indicated the probability of the presence of cancer in the examined bronchus. Single cancer cells or tiny clusters of cells with orangeophilic cytoplasm can appear in specimens obtained from all bronchi, and such cells should not be considered to have originated in the bronchi under examination. Medium-sized or large clusters of cancer cells without degeneration and with basophilic cytoplasm appear only in specimens obtained from bronchi in which a cancer lesion exists, and thus they should be considered to have originated in the bronchi under examination. Cancer cells with orangeophilic cytoplasm in clusters should be considered to have originated in unknown locations. To determine the origin of such cells, one must compare the specimens with those obtained from other segmental and subsegmental bronchi. Our findings suggest that selective brushing of all segmental and subsegmental bronchi is a useful method of detecting unrecognizable second cancers and that the method should be employed for all patients with positive sputum cytology.

Clinicopathological analysis of 19 cases of isolated carcinoma in situ of the bronchus.

Nineteen cases of isolated squamous cell carcinoma in situ (CIS) of the bronchus were described clinicopathologically from among 149 male heavy smokers with roentgenographically occult lung cancer discovered mainly by mass screening performed from 1982 to 1991. All 19 patients had positive sputum cytology tests and negative chest x-ray films and underwent lobectomy (except one who had segmentectomy because of poor lung function). Prior to operation, localization was accomplished by one to eight bronchoscopies using repetitive brush cytology and biopsy. Five cases were bronchoscopically invisible. Polypoid protuberance was noted in three cases, micronodular swelling in three, thickening of spur in five, and mucosal granularity in three. Histology by serial block sectioning showed that there was no nodal involvement in any cases; the maximum length or diameter was 12 mm. Thirteen tumors were < or = 4 mm, four of which were confined to the spur where they occurred. Follow-up data showed a favorable prognosis. Segmentectomy or sleeve resection of bronchus without mediastinal lymph node dissection may be adequate for CIS.

Relation between bronchoscopic findings and tumor size of roentgenographically occult bronchogenic squamous cell carcinoma.

A total of 105 lesions in 98 patients with roentgenographically occult bronchogenic squamous cell carcinoma were examined. The relationship of bronchoscopic findings to the depth of invasion into the bronchial wall and the length of longitudinal extension along the bronchus was documented. From viewpoints of the degree of difficulty of bronchoscopic detection and with reference to the height of the lesions, the bronchoscopic findings were classified into three categories: remarkable, minute, and hidden. Of the 105 lesions, 55 (52%) were remarkable, 27 (26%) were minute, and the remaining 23 (22%) were hidden. Of the 23 hidden lesions, 12 were within and 11 were beyond the range of endoscopic visibility. The maximal depth of bronchial invasion (mean +/- standard error) was 3.07 +/- 0.40 mm in the category designated remarkable and 1.62 +/- 0.47 mm in the category designated minute. The depth was 0.93 +/- 0.36 mm in the hidden lesions within the range of endoscopic visibility and 0.78 +/- 0.21 mm in the hidden lesions beyond the range of endoscopic visibility. The maximal length of longitudinal extension along the bronchus was 19.6 +/- 1.5 mm in the remarkable lesions, 9.9 +/- 1.4 mm in the minute lesions, 5.5 +/- 1.0 mm in the hidden lesions within the range of endoscopic visibility, and 8.6 +/- 2.1 mm in the hidden lesions beyond the range of endoscopic visibility. It is useful for predicting the depth of invasion to classify bronchoscopic findings into these three categories for the study of roentgenographically occult bronchogenic squamous cell carcinomas.

Diagnostic value of differential brushing of all branches of the bronchi in patients with sputum positive or suspected positive for lung cancer.

In roentgenographically occult lung cancer, it is often difficult to determine the location of the tumor despite the existence of cancer. This complicates diagnosis and points to a need for a more systematic method of examination. Differential brushing was performed on all the respective segmental bronchi in both lungs of 196 patients with positive or suspected positive indications of lung cancer as revealed by sputum cytology. Fifty-nine borderline lesions in 43 cases and 107 lung cancer lesions in 95 cases were diagnosed. Localization was possible in 70.4% of the cases. The diagnosis of borderline lesions was also possible. At the first examination, the rate of localization, as compared with that in the historical control group, improved from 64.1% to 95.8%, and, in particular, an improvement from 0% to 86.2% was noted in those cases in which abnormal bronchoscopic findings were not observed. Concurrent multiple primary cancer was also diagnosed in 12.6% of lung cancer cases before treatment. With this method, cytologic findings in sputum and in specimens obtained by brushing and histologic findings of resected lung can be compared in an integrated manner, and henceforth more accurate diagnostic criteria can be established.

[Resected roentgenographically occult bronchogenic squamous cell carcinoma tumor size, survival and recurrence].

The relationship between tumor size and nodal involvement of resected roentgenographically occult squamous cell carcinoma in 127 cases was documented. Survival and recurrent patterns were analyzed. Intrabronchial invasion was observed in 103 cases and extrabronchial invasion in 24 cases. One hundred and nineteen cases (94%) had N0 diseases, six (5%) N1 diseases and two (2%) N2 diseases. One hundred and one cases were in early stage and 26 in non-early stage. Nodal involvement was observed in two (2%) of the 103 cases with intrabronchial invasion and in six (25%) of the 24 cases with extrabronchial invasion. Nodal involvement was noted in none (0%) of 55 cases in whom longitudinal extension of tumors was within 10mm, but was noted in four (9%) of 46 cases in whom it was 11 to 20mm and in four (15%) of 26 cases in whom it was 21 to 55mm. Death from primary lung cancer occurred in three (12%) of the non-early cases, but in none (0%) of the early cases. Death from multiple metachronous lung cancer occurred in one (4%) of the non-early cases and in three (3%) of the early cases. Nodal and extrabronchial involvement reduced survival. Recurrence often involved hilar, mediastinal, supraclavicular nodes, and surgical margin of bronchus.

Rapid expression of collagen type X gene of non-hypertrophic chondrocytes in the grafted chick periosteum demonstrated by in situ hybridization.

We investigated the spatiotemporal localization of collagen Type I, II, and X mRNAs in the subcutaneously grafted chick periosteum by in situ hybridization. Five days after transplantation, we noted three types of histological findings in the grated tissue. (a) Developing trabecular bone exhibited proliferation of spindle-shaped fibroblastic cells and polygonal osteoblasts with moderate signals for collagen Type I mRNA. (b) Developing cartilage contained ovoid chondrocytes with a moderate level of both collagen Type I and II mRNAs. Differentiating chondrocytes with increased collagen Type X mRNA developed during the course of endochondral ossification. (c) An atypical mass of cartilage weakly stained with alcian blue was composed of a large number of non-hypertrophic chondrocytes exhibiting high signals for collagen Type X mRNA. At Day 9, we observed the typical histological features of both membranous and endochondral ossification. However, sparsely distributed chondrocytes with high signals for collagen Type X mRNA were also demonstrated in osteoid and/or woven bone. The phenotype of chondrocytes showing rapid expression of collagen Type X gene derived from grafted periosteum seems to participate in the important role of endochondral bone formation in the early stage of fracture repair.

Lesions preceding squamous cell carcinoma of the bronchus and multicentricity of canceration--serial slicing of minute lung cancers smaller than 1 mm.

A total of ten minute squamous cell carcinomas smaller than 1 mm were found in surgical lung specimens from 108 patients who had roentgenographically occult lung cancer. These minute lesions were detected by submitting, in all the 108 specimens, the whole bronchial tree to 2-mm-thick sequential transverse slicing which was then followed by microscopic examination of each slice on an H-E stained section. When a focus of minute carcinoma was found, the slice was further serially sectioned to study whether there were such carcinoma-related lesions as dysplasia or other atypical changes of epithelia, and when there were, the spatial relation of these with the carcinoma. It was demonstrated that all the minute carcinomas were closely associated with either dysplasia or what we call "basal cells with marked atypia", cells with markedly enlarged nuclei arranged in linear fashion on the basement membrane. The contiguity of these changes with minute carcinoma strongly suggested that they are lesions preceding overt carcinoma. Also, there were some minute foci of carcinoma, which, though not involving the entire epithelial thickness, proved to have already begun microinvasion.

An improved method of bronchial stump closure for prevention of bronchopleural fistula in pulmonary resection.

We performed 880 pulmonary resections from January 1982 to June 1988 using Sweet's procedure for closure of the bronchial stump, in which 39 patients (4.4%) developed bronchial fistulas. Bronchoscopic studies showed that bronchopleural fistulas were located mainly at the corner of the stump. This indicates that the corner is the point with the highest tension when Sweet's procedure is employed. In some cases, stumps were injured by suture materials, resulting in bronchopleural fistulas. Since July 1988, bronchial stumps have been closed by using two pairs of teflon pledgets with additional interrupted sutures. From July 1988 to April 1990, 288 patients were treated by this new method, and only one of them developed bronchopleural fistula. This new method prevents injury of the stump by suture material and reduces the tension at the bronchial stump for a long time. Thus, pulmonary resections can be safely employed even after anti-cancer chemotherapy and/or radiation therapy.