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.

Relationship of lymph node metastasis to primary tumor size and microscopic appearance of roentgenographically occult lung cancer.

We investigated the relationship of lymph node metastasis to primary tumor size and microscopic appearance in 92 resected specimens obtained from patients with roentgenographically occult lung cancer (ROLC) located at a site along the airway between the main bronchus and the sub-subsegmental bronchi. Most of the patients were discovered by mass screening. All were treated surgically after bronchoscopic localization of cancer. The bronchial tree of the resected specimens was serial-sectioned into 2-mm thick blocks from the margin of resection to the sub-subsegmental bronchi. Bronchial wall invasion was noted in some blocks of all the specimens. The length of longitudinal extension (LLE) was defined as the product of the thickness and the number of consecutive blocks involved, counting from the most proximal to the most distal block. LLE was used as primary tumor size. Hilar and mediastinal lymph nodes were examined in 84 patients who underwent lymph node dissection. No nodal involvement was found in 59 cancers with LLE of less than 20 mm. Of 25 cancers with LLE of 20 mm or more, six showed nodal involvement. Eleven in situ carcinomas and four cancers of the "suspicious for invasion" type showed no lymph node metastasis. We contend that no lymph node dissection is required when pulmonary resection is performed for patients with ROLC if it is in situ carcinoma, if it is of the "suspicious for invasion" type, or if the LLE is smaller than 20 mm.

Relationship between length of longitudinal extension and maximal depth of transmural invasion in roentgenographically occult squamous cell carcinoma of the bronchus (nonpolypoid type).

This study was designed to verify our hypothesis that there are two different growth types in roentgenographically occult squamous cell carcinoma of the bronchus. Serial blocks prepared from the entire bronchial tree of 83 resected specimens of occult carcinoma were used for the evaluation of the relationship between the length of longitudinal extension and the maximal depth of transmural invasion. We prepared a length-depth diagram of 92 lesions, including multifocal carcinomas, which confirmed that there are at least two types: Most of these lesions are of the creeping type, which shows a marked superficial growth; the minority are of the penetrating type, which shows a marked downward growth. The diagram suggests that occult carcinoma has a propensity either for longitudinal growth along the bronchial lumen or for transmural growth into the bronchial wall at the time of occurrence. It is likely that the penetrating type grows rapidly and becomes advanced in a short time. Identification of longer lesions of the creeping type is occasionally problematical both at bronchoscopy and at surgical treatment. The stump is usually positive for carcinoma unless frozen sections or imprint specimens of the margin of resection are examined, because it is frequently difficult to identify the proximal end of extension by bronchoscopy.

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.

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.

Results of surgical treatment for roentgenographically occult bronchogenic squamous cell carcinoma.

Ninety-four patients with roentgenographically occult bronchogenic squamous cell carcinoma had surgical resection. Fifty-three reported having no symptoms. In 83 carcinoma was detected by cytologic examination of the sputum during lung cancer screening. The carcinomas were located in segmental bronchi (34 cases), subsegmental bronchi (19 cases), divisional bronchi (17 cases), and subsubsegmental or more peripheral bronchi (15 cases). The number of cases classified by TNM staging were 16 Tis N0 M0, 72 T1 N0 M0, 4 T1 N1 M0, and 2 T2 N1 M0. Extrabronchial invasion of the resected carcinoma was observed in 17 lesions (16 cases). Five of six patients with lymph node metastasis in the resected specimens had carcinoma with extrabronchial invasion. Multiple primary lung cancers were observed in nine patients at the time of operation and in seven subsequently. Four of seven patients with subsequent primary lung cancer had surgical resection, and no recurrence was observed after the second operation. There were two deaths from lung cancer: One was caused by subsequent primary lung cancer and the other by mediastinal lymph node metastasis. In the 75 patients with intrabronchial cancer invasion and without lymph node metastasis who had complete resection, there was no local recurrence or metastasis of cancer. The 5-year survivals were 80.4% (death from all causes) and 93.5% (death from lung cancer). Although subsequent primary lung cancer is troublesome, operation is a reliable treatment for occult bronchogenic squamous cell carcinoma.

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.

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.

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.

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.

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.

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.

Cytologic study of tissue repair in human bronchial epithelium.

The cytologic findings of atypical cells considered to be tissue repair cells after mechanical injury to the bronchial epithelium are reported. These cells were studied in sequential bronchial brushing smears from patients who underwent repeated bronchoscopies for the diagnosis of lung cancer. The cellular findings varied according to the length of time since the previous bronchial brushing. Many cell clusters of highly atypical cells in two-dimensional sheets with large nuclei and prominent nucleoli were observed in specimens taken two or three days after a previous brushing; mitotic figures were observed on day two. In specimens taken on days four and five, the number of atypical cells was decreased and the degree of atypia was slight.

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.

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.

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.

[Experience of Nd-YAG laser treatment in roentogenographically occult bronchogenic squamous cell carcinoma].

Out of 201 patients with roentogenographically occult bronchogenic squamous cell carcinoma, 165 lesions underwent surgical operations, while 41 lesions underwent non-invasive therapy. The 5-year survival rate including all causes of death, was significantly higher in the surgical operation group than that in the non-invasive therapy group (83% vs 43%). Among 14 cases who received Nd-YAG laser treatment, one case died but 4 cases are alive bearing cancer. Since time span of follow-up period is short, it is hard to say that the prognosis of the Nd-YAG laser therapy in fair. Nd-YAG laser treatments were effective for cancers which did not invade beyond bronchial cartilage. From the bronchoscopic examinations and studies with resected lungs, we suggest that Nd-YAG laser treatments may be effective when cancer is extended within 10 mm wide and has only a slight findings in bronchoscopy.

[Surgical treatment of pathological stage III and IV lung cancer less than 3 cm in diameter].

We analyzed stage III and IV lung cancer with tumor size smaller than 3.0 cm. The percentage of adenocarcinoma among the patients with stage III A lung cancer was high. In survival rate, there was no observable difference between the patients with tumor size smaller than 3.0 cm and the patients with tumor size larger than 3.1 cm. But the ratio of the people who had a long survival was high in the latter group. Among the stage IV patients, the pm 1 group with N0 or N1 had a good prognosis (52%, 50% at 5 years).

[T4 advanced lung cancer: results of surgical treatment and indications of surgical resection].

A total of 1,289 patients with primary lung cancer were surgically treated at our hospital from January 1953 to December 1985. Surgical treatment for T4 lung cancer was studied in 93 patients who had pulmonary resections. The relationships between histologic type, stage, method of resection, curability, nodal involvement, pleural involvement, site of invasion, pleural metastasis, pleural effusion, combination therapy, and the survival rate were analyzed. The survival rate of 93 patients with T4 lung cancer was 17% at 3 years and 7% at 5 years. Three-year survival rate of 39 patients with adenocarcinoma, 34 patients with squamous cell carcinoma, and 9 patients with large cell carcinoma was 7%, 23%, and 14%, respectively. Two-year survival rate of 6 patients with small cell carcinoma was 17%. Four-year survival rate of 14 patients who had complete resection was 33%. On the other hand, four-year survival rate of 77 patients who had incomplete resection was 7%. Three-year survival rate of 6 patients with N0 disease, and 19 patients with N1 disease, 46 patients with N2 disease, and 22 patients with NX disease was 40%, 39%, 0%, and 15%, respectively. Two patients, who had partial resection of the left atrium because carcinoma made an invasion upon it, had survived more than 5 years. All patients with esophageal invasion or tracheal invasion had died within a year. Indications of surgical resection for patients with T4 lung cancer should be limited to patients with N0 and N1 disease. Radical pulmonary resection can be performed in patients who are expected to have complete resection.

[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.