Dynamic viscoelasticity of protease-treated rice batters for gluten-free rice bread making.

Papain (cysteine protease), subtilisin (Protin SD-AY10, serine protease), and bacillolysin (Protin SD-NY10, metallo protease) increased the specific volume of gluten-free rice breads by 19-63% compared to untreated bread. In contrast, Newlase F (aspartyl protease) did not expand the volume of the rice bread. In a rheological analysis, the viscoelastic properties of the gluten-free rice batters also depended on the protease categories. Principal component analysis (PCA) analysis suggested that the storage and loss moduli (G' and G″, respectively) at 35 °C, and the maximum values of G' and G″, were important factors in the volume expansion. Judging from the PCA of the viscoelastic parameters of the rice batters, papain and Protin SD-AY10 improved the viscoelasticity for gluten-free rice bread making, and Protin SD-NY effectively expanded the gluten-free rice bread. The rheological properties differed between Protin SD-NY and the other protease treatments.

Novel substrate specificities of two lacto-N-biosidases towards ß-linked galacto-N-biose-containing oligosaccharides of globo H, Gb5, and GA1.

We describe the novel substrate specificities of two independently evolved lacto-N-biosidases (LnbX and LnbB) towards the sugar chains of globo- and ganglio-series glycosphingolipids. LnbX, a non-classified member of the glycoside hydrolase family, isolated from Bifidobacterium longum subsp. longum, was shown to liberate galacto-N-biose (GNB: Galß1-3GalNAc) and 2'-fucosyl GNB (a type-4 trisaccharide) from Gb5 pentasaccharide and globo H hexasaccharide, respectively. LnbB, a member of the glycoside hydrolase family 20 isolated from Bifidobacterium bifidum, was shown to release GNB from Gb5 and GA1 oligosaccharides. This is the first report describing enzymatic release of ß-linked GNB from natural substrates. These unique activities may play a role in modulating the microbial composition in the gut ecosystem, and may serve as new tools for elucidating the functions of sugar chains of glycosphingolipids.

Stratifying the risk of lymph node metastasis in undifferentiated-type early gastric cancer.

AIM: To study how lymph node metastasis (LNM) risk is stratified in undifferentiated-type early gastric cancer (undiff-EGC) dependent on combinations of risk factors. METHODS: Five hundred and sixty-seven cases with undiff-EGC undergoing gastrectomy with lymphadenectomy were examined retrospectively. Using clinicopathological factors of patient age, location, size, an endoscopic macroscopic tumor form, ulceration, depth, histology, lymphatic involvement (LI) and venous involvement (VI), LNM risk was examined and stratified by conventional statistical analysis and data-mining analysis. RESULTS: LNM was positive in 44 of 567 cases (7.8%). Univariate analysis revealed > 2 cm, protrusion, submucosal (sm), mixed type, LI and VI as significant prognostic factors and > 2 cm and LI-positive were independent factors by multivariate analysis. In preoperatively evaluable factors excluding LVI, sm and > 2 cm were independent factors. According to the depth and size, cases were categorized into the low-risk group [m and ≤ 2 cm, 0% (LNM incidence)], the moderate-risk group (m and > 2 cm, 5.6%; and sm and ≤ 2 cm, 6.0%), and the high-risk group (sm and > 2 cm, 19.3%). On the other hand, LNM occurred in 1.4% in all LI-negative cases, greatly lower than 28.2% in all LI-positive cases, and LNM incidence was low in LI-negative cases even in the moderate- and high-risk groups. CONCLUSION: LNM-related factors in undiff-EGC were depth and size preoperatively while those were LI and size postoperatively. Among these factors, LI was the most significantly correlated factor.

1,3-1,4-α-L-fucosynthase that specifically introduces Lewis a/x antigens into type-1/2 chains.

α-L-fucosyl residues attached at the non-reducing ends of glycoconjugates constitute histo-blood group antigens Lewis (Le) and ABO and play fundamental roles in various biological processes. Therefore, establishing a method for synthesizing the antigens is important for functional glycomics studies. However, regiospecific synthesis of glycosyl linkages, especially α-L-fucosyl linkages, is quite difficult to control both by chemists and enzymologists. Here, we generated an α-L-fucosynthase that specifically introduces Le(a) and Le(x) antigens into the type-1 and type-2 chains, respectively; i.e. the enzyme specifically accepts the disaccharide structures (Galß1-3/4GlcNAc) at the non-reducing ends and attaches a Fuc residue via an α-(1,4/3)-linkage to the GlcNAc. X-ray crystallographic studies revealed the structural basis of this strict regio- and acceptor specificity, which includes the induced fit movement of the catalytically important residues, and the difference between the active site structures of 1,3-1,4-α-L-fucosidase (EC 3.2.1.111) and α-L-fucosidase (EC 3.2.1.51) in glycoside hydrolase family 29. The glycosynthase developed in this study should serve as a potentially powerful tool to specifically introduce the Le(a/x) epitopes onto labile glycoconjugates including glycoproteins. Mining glycosidases with strict specificity may represent the most efficient route to the specific synthesis of glycosidic bonds.

[The usefulness of adjuvant therapy using gamma knife radiosurgery for the recurrent or residual nonfunctioning pituitary adenomas].

We evaluated the treatment results of nonfunctioning pituitary adenomas in the era of radiosurgery. Between January 1994 and December 2003, we operated on 44 patients with nonfunctioning pituitary adenomas. 43 patients were operated on by transsphenoidal surgery and one patient was operated on by the transcranial approach. Total removal was able to be achieved in 13 patients (30%). Gamma knife radiosurgery was performed for residual tumor in 26 patients and for recurrence in 2 patients. The mean tumor diameter at the gamma knife radiosurgery was 18.2 mm (7.9 to 26.3 mm). The treatment dose was a mean of 12.3 Gy (8 to 16 Gy) to the tumor margin. The mean follow-up period after radiosurgery was 36.4 months. Tumor growth control was able to be achieved in 26 patients (93%). Two patients (7%) required adrenal and thyroid hormonal replacement during the follow-up period after radiosurgery due to radiation-induced endocrinopathy. None of the patients suffered from new cranial nerve deficits. This included optic neuropathy. Surgical resection using transsphenoidal surgery and subsequent gamma knife radiosurgery for residual and recurrent tumor proved to have a highly effective tumor growth control rate, and maintained the quality of life in patients with nonfunctioning pituitary adenomas.

Radiosurgery for pituitary metastases.

Gamma knife radiosurgery (GKS) was used to treat seven patients with pituitary metastases between November 1994 and February 2003. The diagnoses were based on magnetic resonance imaging and clinical symptoms in six patients and by previous surgery in one patient. The cancer originated in the lung in five patients, and in the breast in two patients. The tumor volume was 0.2 to 9.6 cm3 (mean 4.0 cm3). The marginal dose was 10 to 14 Gy (mean 11.9 Gy) because of the close proximity to the optic apparatus. The maximum radiation dose to the optic apparatus was 8 to 10 Gy (mean 9.5 Gy). The survival period after GKS was 0.3 to 42 months (mean 11.5 months). Five patients died of systemic disease, and one patient died of unknown causes 10 days after GKS. Tumor growth was controlled in five of the six patients (83%) followed up after GKS. Tumor regrowth was seen 18 months after GKS in one patient. The clinical symptoms improved in five of the six patients (83%) followed up. GKS is effective and useful for the primary treatment of pituitary metastases with limited survival and less invasiveness compared to conventional radiation therapy.

[The treatment for asymptomatic meningiomas in the era of radiosurgery].

We evaluated the treatment results in asymptomatic meningiomas in the era of radiosurgery. We encountered 56 patients with asymptomatic meningiomas from January, 1994 to December, 2001. There were 16 male patients and 40 female patients. The most common location was the cerebral convexity. We performed gamma knife radiosurgery for three patients after diagnosis of cavernous sinus meningiomas. The other patients were followed-up after diagnosis for a mean follow-up period of 30.8 months (5 months to 8 years). Tumor growth was verified in 25 patients (43%) and two patients (3.7%) suffered neurological deficits. Among the tumor growth patients, operative resection was performed in six patients (11%) and gamma knife radiosurgery was performed in four patients (7.5%). The other patients were placed under observation. We decided the treatment strategy paying attention to the tumor location, tumor growth speed, tumor size and operative difficulty. The tumor growth rate was 0.1 cm/year among the patients with tumor growth. It was 0.72 cm/year (0.02 to 1.54 cm/year) in the operative group, 0.3 cm/year (0.04 to 0.76 cm/year) in the gamma knife radiosurgery group and 0.08 cm/year (0.01 to 0.76 cm/year) in the observed group. One patient suffered procedure-related deterioration of paresis after operation. The treatment of asymptomatic meningiomas must be decided depending on their natural history and locations. We recommend early radiosurgery for cavernous sinus meningiomas.

[The surgical treatment strategy and results of parasellar meningiomas in the era of radiosurgery].

We evaluated the surgical treatment results of parasellar meningiomas in the era of radiosurgery. We treated 24 patients of parasellar meningiomas surgically. The median age was 60 yrs (ranging from 29 to 82 yrs). The most common tumor location was the sphenoid ridge in 12 patients and the tuberculum sellae in 7 patients. The pterional approach using fronto-temporal craniotomy was performed for all patients. The residual or recurrent tumors were treated by gamma knife radiosurgery (GKS). We are able to follow up these cases for a median of 3.8 yrs (ranging from 1 to 8 yrs) after the operations. The radicality of tumor resection was Simpson grade II in 13 patients (54%), grade III in 3 patients (13%) and grade IV in 7 patients (33%). Clinical improvement was achieved in 81% of the patients. Of the patients who had visual disturbance preoperatively, 8 patients (73%) showed improvement, but 3 patients suffered deterioration postoperatively. None of the patients died. One patient suffered transient memory disturbance and one patient suffered mild facial numbness postoperatively. Boost radiosurgery for the residual tumors was performed for six patients and tumor growth control was able to be achieved in all patients, with a median of 3.1 years follow-up period (ranging from 0.5 to 6 yrs). Five patients with tumor regrowth or recurrence were treated by GKS. We recommend fronto-temporal craniotomy with nonradical resection for parasellar meningiomas and radiosurgery for residual and recurrent tumors. This strategy will achieve good functional outcome with long-term tumor growth control.