Tissue ionome response to rhizosphere pH and aluminum in tea plants (Camellia sinensis L.), a species adapted to acidic soils.

The growth of tea plants (Camellia sinensis L.) is promoted by the presence of aluminum (Al), a beneficial element under acidic conditions, but the influence of rhizosphere pH on this interaction is not known. To understand the mechanisms underlying the adaptation to acidic rhizosphere conditions, we evaluated ionome profiles and the effect of pH on tea growth in hydroponic culture. The optimum pH for tea growth was around pH 4.2, and growth was inferior under a pH less than 3.8 or higher than 5.0. Under the optimum pH growth and Al accumulation were markedly stimulated by Al treatment. Al content and accumulation in new and mature leaves and new roots (the predominant tissues that accumulate minerals in tea plants) gradually declined with decrease in pH, especially in new roots. Ionome profiles drastically altered Al treatment, but changes were more pronounced in new roots than in new or mature leaves and did not depend on pH. Although the uptake of most cationic minerals in new roots was decreased by Al treatment, cationic mineral contents in new and mature leaves were not decreased by Al. In contrast to other plant species, the content and accumulation of manganese, despite it being a cationic nutrient, were significantly increased by Al treatment. These results indicated that one role of Al as a beneficial element was to maintain the shoot nutrient status by effectively utilizing Al-limited elements in the roots.

Quantitative validation of nicotine production in tea (Camellia sinensis L.).

Endogenous nicotine was confirmed to be present in tea plants (Camellia sinensis L.) by liquid chromatography-tandem mass spectrometry of tea samples from tea-producing regions in six Asian countries. All samples contained nicotine (0.011-0.694 µg g-1 dry weight). Nicotine contents remained constant during manufacturing of green, oolong and black teas, implying that nicotine is stable against heating, drying, enzymatic oxidation and mechanical damage during processing. Flower buds and seeds of cultivar Yabukita also contained nicotine (0.030-0.041 µg g-1 dry weight). A comparison of two cultivars revealed that higher nicotine contents were found in the black tea cultivar Benifuki. All plant parts of hydroponic Yabukita contained nicotine (0.003-0.013 µg g-1 dry weight). Tea cells cultured in B5 medium as well as roots and stems of tea seedlings contained nicotine levels similar to those of new leaves from field-grown plants. Although the levels of endogenous nicotine in tea plants are extremely low and sample contamination cannot be discounted, these levels exceed the maximum acceptable limit in Japan (0.01 µg g-1 dry weight).

[Clinical Study of Percutaneous Radiofrequency Ablation for Small Renal Tumor].

Percutaneous radiofrequency ablation (PRFA) has been applied as an option of minimally invasive treatment for small renal tumor. We retrospectively evaluated 5 patients with small renal tumor for whom PRFA was performed at our hospital. The average age was 69.6 years (range 45-86), average tumor diameter 20.0 mm (8-34), average preoperative glomerular filtration rate (eGFR) 66.8 ml/min/1.73m2 (42. 1-93.2), and follow-up period was 23.4 months (8-34). There were two minor complications (grade 1), including perirenal hematoma and pain at the probe insertion site in each patient. One patient had local recurrence 22 months after PRFA and the additional PRFA will be performed. PRFA could be a feasible, safe and effective therapy for small renal tumor.

[Modified Toyoda Method for Complete Duplex Ureters in Tubeless Cutaneous Ureterostomy : Report of a Case].

A 65-year-old man underwent radical cystectomy and cutaneous ureterostomy for bladder cancer. Because the patient had left complete duplex ureters,we modified the Toyoda method to construct the tubeless cutaneous ureterostomy with a unilateral stomal creation. The adhesive portion of both ureteral walls in duplex ureters was simultaneously cut approximately 5 mm from the distal end longitudinally,and the common sheath of the cut portion of the ureters was cut approximately 5 mm longitudinally in order to separate the two ureters. This procedure was repeated to make a"fish-mouth"aperture. Two opened ureteral walls were formed into the upper and lower plates of the Toyoda method stoma,respectively. To our knowledge,this is the first report of a surgical procedure of the tubeless cutaneous ureterostomy in complete duplex ureters.

Development of Primary Thyroid Lymphoma during an Ultrasonographic Follow-up of Hashimoto's Thyroiditis: A Report of 9 Cases.

We herein experienced 9 patients with primary thyroid lymphoma that developed during 3-18 years of ultrasonographic follow-up of Hashimoto's thyroiditis. All nine patients had localized mucosa-associated lymphoid tissue (MALT) lymphoma. Two patients had diffuse type, one had mixed type, and six had nodular type according to the ultrasonographic classification. A clearly enlarging goiter was observed before the diagnosis of lymphoma in 3 patients. An enlarging goiter was not apparent in the remaining 6 patients with nodular type lymphoma, however, the emergence or enlargement of a hypoechoic nodular lesion was observed. Thyroid MALT lymphoma may be diagnosed early by an ultrasonographic follow-up of Hashimoto's thyroiditis.

Value of thyroid specific peroxidase and Ki-67 stains in preoperative cytology for thyroid follicular tumors.

BACKGROUND: The aim of this study was to elucidate immunocytochemically whether thyroid specific peroxidase (TPO) and Ki-67 can complement fine-needle aspiration (FNA) cytology as useful markers in order to distinguish between follicular adenoma (FA) and follicular carcinoma (FC). METHODS: We studied 40 FAs and 68 FCs obtained by surgical resection. FNA cytology smears were divided into two groups: Cytology-A (Cy-A) (94 cases) with typical benign cytology and Cytology-B (Cy-B) (14 cases) with atypical cytology. FCs were divided into two groups: FC-I (42 cases) without any poorly differentiated structures and FC-II (26 cases) with some poorly differentiated structures. Cytology smears and histology from FAs and FCs were studied immunocytochemically for thyroid specific peroxidase (TPO) and Ki-67. RESULTS: TPO expression was negative in 12.5% FAs, 21.4% FC-I, and 46.2% FC-II. In 68 FC cases, Cy-B were more frequently observed in TPO-negative cases (38.1%) than in TPO-positive cases (12.8%). The mean Ki-67 LI was 0.46 in FAs, 0.53 in FC-I, and 1.13 in FC-II. The high Ki-67 LI was correlated with Cy-B. Moreover, higher Ki-67 LI showed a close relationship with distant metastasis. In 94 Cy-A cases, 54 cases were FCs. When 38 cases with negative TPO or Ki-67 LI over 0.62 were extracted from them, as many as 28 cases were FCs, the rate of FCs were significantly higher than the rest. CONCLUSION: Therefore, addition of TPO stain and Ki-67 stain to routine Papanicolaou stain could improve the diagnostic reliability of FNA cytology for FC with high degree of malignancy.

Follicular thyroid cancer in children and adolescents: clinicopathologic features, long-term survival, and risk factors for recurrence.

Children and adolescents represent 1-1.5% of all patients with thyroid cancer (TC). The vast majority of TC in children and adolescents is papillary TC; follicular TC (FTC) is exceedingly rare. In this study, we evaluate the clinical and pathological features of FTC in children and adolescents. We also report the risk factors for post-operative tumor recurrence and the associated outcomes. Twenty children and adolescents (under 21 years old) with FTC have been treated and followed at Noguchi Thyroid Clinic and Hospital Foundation since 1946. All patients underwent surgery (lobectomy, 11; subtotal thyroidectomy, 8; and total thyroidectomy, 1), and 8 patients received postoperative external beam radiation therapy. The incidence of FTC in children and adolescents was 1.9% among all FTC patients treated in our hospital. Histopathology revealed vascular and capsular invasion in 9 and 20 patients, respectively. The tumor recurrence rate in FTC with vascular invasion is significantly higher than in those without it (p = 0.038). No other factors were significant. Patients with recurrences were treated with completion thyroidectomy and ¹³¹I radioactive iodine therapy. There were no significant differences in the rates of disease-free survival or cause-specific survival when pediatric/adolescent FTC patients were compared to adults with FTC. FTC is very rare among children and adolescents, but the outcomes are similar to those observed among adults. Vascular invasion is poor prognostic indicator in pediatric/adolescent FTC patients.

Remission of anorexia nervosa after thyroidectomy: A report of two cases with Graves' disease and anorexia nervosa.

We report two patients with anorexia nervosa and Graves' disease who received subtotal thyroidectomy for Graves' disease and concomitantly experienced remission from anorexia nervosa. Both were young women (aged 20 and 26) at the time of surgery. Both had well controlled thyroid function and eating behavior at the time of surgery. Both were followed for over five years without relapse of anorexia nervosa or hyperthyroidism. These cases suggest the existence of an endocrine factor originating from the thyroid gland that is involved in the pathogenesis of anorexia nervosa. Since patients of thyroidectomy can remain in good health with supplement of thyroxine alone, it can be hypothesized that this anorexigenic endocrine factor is an evolutionary relic not necessary for the normal function of humans and does not have physiological effects unless secreted beyond normal levels. Given that, it implies the existence of a creature in the animal kingdom for which such an anorexigenic hormone is essential for survival. Migrating birds eat beyond their caloric expenditure before migration and become anorexic for the duration of their flight. It is also known that their thyroid function is elevated during migration. The normal physiology of migration is a complex mechanism involving the hypothalamic, pituitary, thyroid, adrenal and reproductive hormones. The mechanism of disease, however, can be simpler. A review of the literature is presented that suggest a heretofore unreported thyroid hormone, which is involved in the regulation of migration behavior, may be the responsible factor behind anorexia nervosa.

Improving the diagnostic accuracy of thyroid follicular neoplasms: cytological features in fine-needle aspiration cytology.

Cytological diagnosis in follicular neoplasms of the thyroid has to surmount some difficulties. Capsular/vascular invasions or metastasis are the histological criteria for follicular carcinoma (FC), and, on fine-needle aspiration (FNA) samples, marked cytological atypias are only observed in moderately to poorly differentiated FC, while they may be completely lacking in well differentiated angio- or capsulo-invasive FC. To clarify the cytological features and to improve the accuracy and reliability of aspiration cytology, 892 follicular adenomas and 82 FCs were reviewed. A macrofollicular pattern or large sheet pattern of follicular cells with thin colloid in the background were found to be indicators of follicular adenoma. Crowding or irregular arrangement of follicular cells were found to indicate microfollicular lesions but could not discriminate between benign and malignant conditions. High nucleo-cytoplasmic ratio, nuclear atypia, and coarse granular or dense chromatin were more important criteria for malignancy than nuclear grooves or intranuclear cytoplasmic inclusions. The cytomorphologic features of the follicular neoplasms of the thyroid are described, and the difficulties encountered in the cytodiagnosis of follicular lesions are discussed at length.

Modified radical neck dissection is better than partial dissection of lymph nodes.

BACKGROUND: For papillary thyroid cancer, lymph node excision may be less important for smaller tumors and rather limited node excision might be sufficient to prevent recurrence. METHODS: We compared patients who had undergone modified radical neck dissection with patients who had undergone limited node basin excision. Between 1967 and 1996, 4966 patients at our institution had resection of pure papillary carcinoma tumors larger than 11 mm. RESULTS: In patients whose tumors were larger than 25 mm without local adhesion, modified radical neck dissection had a significantly better prognosis compared to that of more limited node excision. For patients with a primary tumor larger than 11 mm, modified radical had a better prognosis than partial node excision only if there was adhesion or invasion to the recurrent nerve, jugular vein, common carotid artery, vagus nerve, or trachea. CONCLUSION: The size of papillary cancer of the thyroid directly affects prognosis and the role of nodal dissection.

Successful treatment of anaplastic thyroid carcinoma with a combination of oral valproic acid, chemotherapy, radiation and surgery.

Anaplastic thyroid carcinoma (ATC) is the most aggressive of thyroid cancers whose treatment is not yet established and mortality is extremely high. Recent in vitro studies have shown that valproic acid (VA), a newly identified histone deacetilase (HDAC) inhibitor, induces apoptosis, modulates differentiation gene expression of thyroid tumors and enhances the sensitivity of anaplastic cancer cell lines to doxorubicin. We report a case of successful treatment of anaplastic thyroid carcinoma with a combination of oral valproic acid, chemotherapy consisting of cisplatin and doxorubicin, external and intra-operative radiation and surgery. Tumor volume decreased by 50.7% under CT measurement and 44.6% under sonogram measurement over the course of the treatment. No significant rebound of tumor size was observed between each cycle of chemotherapy. Serial cytology performed via fine needle aspiration (FNA) presented a rapidly changing profile of cell types, starting with anaplastic and proceeding through increasingly well differentiated presentations. Only microscopic remnants of ATC cells were found in the histological examination of the resected thyroid. Ga scintigraphy and whole body PET scan six months after surgery revealed no evidence of recurrence or metastasis. As of Nov. 22, 2008, the patient is alive and disease free two years after diagnosis.

Papillary microcarcinoma.

BACKGROUND: Papillary microcarcinoma (PMC) is increasing in incidence because of diagnosis by ultrasound-guided fine-needle aspiration cytology. METHODS: Between January 1966 and December 1995, we treated 6019 patients with papillary cancer; among them, 2070 patients with PMC were studied. RESULTS: PMC is essentially very similar to papillary cancer that is 11 mm or larger and has a very good prognosis. Smaller tumors and younger patients have a better prognosis. Among PMC, larger tumors (6-10 mm) recur in 14% at 35 years compared with 3.3% in patients with smaller tumors. Patients older than 55 years have recurrence in 40% at 30 years, with a worse prognosis than younger patients who have a recurrence rate of less than 10%. Extracapsular invasion by the primary tumor also has a higher recurrence rate. The majority of recurrences are in the neck. Therefore, annual ultrasound of the neck is effective for recurrence surveillance. CONCLUSION: Papillary microcarcinoma is similar to larger papillary carcinomas with tumor characteristics and age-based recurrence rate that extends for many years, justifying long surveillance after surgery.

Mutational analysis of the APC gene in cribriform-morula variant of papillary thyroid carcinoma.

INTRODUCTION: Familial adenomatous polyposis (FAP) is an inherited autosomal dominant syndrome caused by germline mutations in the adenomatous polyposis coli (APC) gene. Papillary thyroid cancer is one of the extracolonic manifestations of FAP. A characteristic histologic feature of this type of thyroid tumor is the cribriform-morula variant of papillary thyroid carcinoma (CMVPTC). METHODS: To investigate roles of the APC and beta-catenin genes in the development of CMVPTC, we examined germline and somatic mutations of these genes in a female patient with CMVPTC and FAP. The patient had undergone total colectomy at the age of 19 years and total thyroidectomy at age 25 years. RESULTS: Numerous tumors were disseminated in both lobes of the thyroid gland, and histopathologic examination revealed typical CMVPTC. DNA was extracted from peripheral blood leukocytes and 12 CMVPTC tumors, and exons 1-15 of the APC gene and exon 3 of the beta-catenin gene were examined. A germline mutation was detected in exon 13 of the APC gene, and this mutation generated a premature stop codon. Six somatic mutations (922delC, 1602delA, 1821delT, 1920delG, 2706del20, 2804insA) were found in the CMVPTC specimens. All mutations were truncating mutations in the N-terminus of the APC protein. Loss of heterozygosity was not observed in the remaining tumor tissues without somatic APC mutations. There were no mutations of the beta-catenin gene in peripheral blood leukocytes or 12 CMVPTC specimens. CONCLUSIONS: These results suggest that APC mutations play an important role in the development of CMVPTC and occur predominantly in the 5' side of the APC gene between codons 308 and 935.

Significance of an insular component in follicular thyroid carcinoma with distant metastasis at initial presentation.

Risk factors for distant metastasis were studied in 82 patients with follicular thyroid carcinoma (FTC). Metastases to either the lung or bone existing at the time of presentation were confirmed by I-131 radio-iodine uptake in 10 patients. FTC with an insular component was found in eight patients. Univariate analysis of 14 possible risk factors showed 7 to be statistically significant: insular component, poorly differentiated carcinoma, trabecular component, serum thyroglobulin level before surgery, patient age at the time of presentation, solid component, and vascular invasion (ranked by p values). After further analysis of the interrelation of the factors and of the logistic regression curves, we concluded that presence of an insular component and patient age were the only independent risk factors. Distant metastasis was not detected in any of the 27 patients < or = 49 yr old. Among the 55 older patients (> or = 50 yr old), 5 of the 49 (10%) without an insular component and 5 of the 6 (83%) with an insular component had distant metastasis. The remaining older patient with an insular component but without distant metastasis showed a gradual increase in thyroglobulin levels after total thyroidectomy.

Thyroid evaluation in patients with primary hyperparathyroidism.

We evaluated the efficacy of preoperative high-resolution ultrasonography (US) for diagnosing possible concomitant thyroid disease which affects the surgical management in patients with primary hyperparathyroidism (pHPT). One hundred and nine patients with sporadic pHPT underwent US with or without ultrasound-guided fine-needle aspiration biopsy (US-FNAB). Diagnosis of concomitant thyroid nodules by US and US-FNAB were compared with the histopathological findings. Of the 109 patients, 19 (17.4%) had malignant thyroid nodules, 26 (23.9%) had benign thyroid nodules alone, and 12 (11.0%) had diffuse goiter. The sensitivity, specificity, and accuracy of diagnosing 72 thyroid nodules were 91.3%, 91.8%, and 91.7% for US, 57.9%, 94.3%, and 81.5% for US-FNAB, and 95.7%, 91.8%, and 93.1% for combined US and US-FNAB, respectively. True positive/false negative ratio of US-FNAB diagnosis was significantly lower in nodules of 5-9 mm than nodules of 10 mm or more. Four unexpected thyroid cancers existed at a different site in 3 of the 39 patients with palpable thyroid disease. Five thyroid cancers were histopathologically confirmed in 5 (7.1%) of 70 patients without palpable thyroid disease. Eight (88.9%) of the 9 non-palpable thyroid cancers were accurately diagnosed by combined US and US-FNAB. Preoperative US is useful for evaluation of possible concomitant thyroid disease, especially for prediction of malignancy.

Immunostaining of galectin-3 and CD44v6 using fine-needle aspiration for distinguishing follicular carcinoma from adenoma.

To evaluate the clinical applicability of galectin-3 and CD44 variant 6 (CD44v6) immunostaining in fine-needle aspiration cytology (FNAC) of thyroid follicular tumors, 79 cytological specimens (35 follicular carcinomas and 44 follicular adenomas) were studied. The positive rates of galectin-3 and CD44v6 were 89 and 74% in follicular carcinoma, respectively, and 25 and 30% in follicular adenoma, respectively. There were no significant correlations between the expression of galectin-3 or CD44v6 in follicular carcinoma and characteristics such as capsular invasion, vascular invasion, metastasis, or tumor size. Positive staining of either galectin-3 or CD44v6 resulted in a diagnostic sensitivity of 97% and a specificity of 52% for follicular carcinoma among follicular tumors. Immunostaining of galectin-3 or CD44v6 using cytological specimens can provide independent information on conventional morphological findings of cytology to distinguish follicular carcinoma from adenoma.

Primary thyroid lymphoma associated with Graves' disease.

We report herein a case of thyroid mucosa-associated lymphoid tissue (MALT) lymphoma in a patient receiving antithyroid drug therapy for Graves' disease. A 75-year-old woman first presented with finger tremor and was diagnosed with Graves' disease on the basis of clinical and laboratory findings. Three years later, she presented with rapid and painless enlargement of the thyroid. Ultrasonography revealed a circumscribed hypoechoic area bilaterally in each lobe of the thyroid, and fine-needle aspiration biopsy showed diffuse monotonous infiltration of small- to medium-sized atypical lymphoid cells. (67)Ga scintigraphy was positive exclusively in the thyroid. After total thyroidectomy, the patient received radiation therapy for treatment of stage IE primary thyroid lymphoma. Results of histological examination, immunohistochemical analysis, and flow cytometric analysis confirmed MALT lymphoma. To our knowledge, there have been few published reports of primary thyroid lymphoma associated with Graves' disease. Our experience with this case, though rare, indicates that an enlarged thyroid in cases of Graves' disease should be examined carefully for primary thyroid lymphoma.

Diagnostic applicability of dipeptidyl aminopeptidase IV activity in cytological samples for differentiating follicular thyroid carcinoma from follicular adenoma.

HYPOTHESIS: Dipeptidyl aminopeptidase IV (DPP IV) activity in cytological samples from a follicular thyroid tumor is the most sensitive and specific indicator for the detection of follicular carcinoma of the thyroid gland. Dipeptidyl aminopeptidase IV activity is independent of cytological characteristics and superior to other clinical findings. DESIGN AND PATIENT SELECTION: Among the patients surgically treated for follicular thyroid tumors, we recruited approximately equal numbers of those with true-positive (n = 19), true-negative (n = 26), false-negative (n = 16), and false-positive (n = 18) cytological characteristics. MAIN OUTCOME MEASURES: We examined DPP IV activity using cytological specimens obtained from 35 patients with follicular thyroid carcinomas and 44 patients with follicular adenomas. Tumor size, patient age, serum thyroglobulin level, and ultrasonographic findings were also analyzed. RESULTS: The positive rate of DPP IV activity was 97% in 35 patients with follicular thyroid carcinomas and 5% in 44 patients with follicular adenomas, resulting in a sensitivity of 97%, a specificity of 95%, and an overall accuracy of 96%. This discriminating ability of DPP IV activity was far higher than that of tumor size, patient age, serum thyroglobulin level, or ultrasonographic findings. CONCLUSIONS: Positive DPP IV activity in cytological samples is the best discriminatory marker between follicular thyroid carcinoma and follicular adenoma. Its application could alter the clinical management of patients with follicular thyroid tumors.

Morphological abstraction of thyroid tumor cell nuclei using morphometry with factor analysis.

Various morphonuclear studies by digital image analysis have successfully been applied to quantify the nuclear morphology, including chromatin distribution pattern, in cytology of various organs; however, the majority of past reports have not shown correlation between the quantitative data by digital image analysis and cytological findings in practical diagnosis. In this report, we present the usefulness of morphological abstraction to combine the objective data and subjective observation in cytological diagnosis. Randomly selected, 100 cells in each Papanicolaou-stained ABC smear samples of 39 benign and malignant thyroid tumor cases were studied. Gray-level image data provided seven parameters for nuclear size, four parameters for nuclear shape, and 16 parameters showing the nuclear chromatin patterns from high-dimensional texture analysis of using co-occurrence and run-length matrices. To statistically abstract nuclear morphology, factor analysis was used. Factor analysis classified morphological nuclear characters as abstraction parameter into five abstract parameters composed of nuclear size, shape, heterogeneity, and contrast and homogeneity of chromatin pattern. The nuclei of papillary carcinoma showed larger size, more irregular shape, and higher contrast of chromatin pattern than those of the benign group. The follicular carcinomas have larger nucleus in each cell and more monotonous chromatin pattern among cells in each case than those of the benign group. Morphological abstraction by morphometry with factor analysis may provide a practical approach to the detection of the underlying characteristics of nuclear morphology in aspiration biopsy cytology.