Prediction of cervical lymph node metastasis from immunostained specimens of tongue cancer using a multilayer perceptron neural network.

BACKGROUND: Although cervical lymph node metastasis is an important prognostic factor for oral cancer, occult metastases remain undetected even by diagnostic imaging. We developed a learning model to predict lymph node metastasis in resected specimens of tongue cancer by classifying the level of immunohistochemical (IHC) staining for angiogenesis- and lymphangiogenesis-related proteins using a multilayer perceptron neural network (MNN). METHODS: We obtained a dataset of 76 patients with squamous cell carcinoma of the tongue who had undergone primary tumor resection. All 76 specimens were IHC stained for the six types shown above (VEGF-C, VEGF-D, NRP1, NRP2, CCR7, and SEMA3E) and 456 slides were prepared. We scored the staining levels visually on all slides. We created virtual slides (4560 images) and the accuracy of the MNN model was verified by comparing it with a hue-saturation (HS) histogram, which quantifies the manually determined visual information. RESULTS: The accuracy of the training model with the MNN was 98.6%, and when the training image was converted to grayscale, the accuracy decreased to 52.9%. This indicates that our MNN adequately evaluates the level of staining rather than the morphological features of the IHC images. Multivariate analysis revealed that CCR7 staining level and T classification were independent factors associated with the presence of cervical lymph node metastasis in both HS histograms and MNN. CONCLUSION: These results suggest that IHC assessment using MNN may be useful for identifying lymph node metastasis in patients with tongue cancer.

Interchanges and movements of humpback whales in Japanese waters: Okinawa, Ogasawara, Amami, and Hokkaido, using an automated matching system.

Humpback whales in the western North Pacific are considered endangered due to their small population size and lack of information. Although previous studies have reported interchanges between regions within a population, the relationship between the geographic regions of a population in Japan is poorly understood. Using 3,532 fluke photo IDs of unique individuals obtained from four areas in Japan: Hokkaido, six IDs (2009-2019); Ogasawara, 1,477 IDs, from two organizations (1) Everlasting nature of Asia (1987-2020) and (2) Ogasawara Whale Watching Association, (1990-2020); Amami, 373 IDs (1992-1994, 2005-2016); Okinawa, 1,676 IDs (1990-2018), interchanges were analyzed. The ID matchings were conducted using an automated system with an 80.9% matching accuracy. Interchange and within-region return indices were also calculated. As a result, number of matches and interchange indices follow locations, Hokkaido-Okinawa (3, 0.31), Amami-Ogasawara (36, 0.06), Amami-Okinawa (222, 0.37), and Okinawa-Ogasawara (225, 0.08), respectively. Interchange indices among Japanese areas were much higher than the indices between Ogasawara/Okinawa and Hawaii (0.01) and Mexico (0.00) reported in previous studies, indicating that the Japanese regions are utilized by the same population. At the same time, the frequency of interchanges among the three breeding areas vary, and the high within-region return indices in respective breeding areas suggest the site fidelity of the whales in each area at some level. These results indicate the existence of several groups within the population which are possibly be divided into at least two groups based on geographical features: one tend to utilize Ogasawara and the Mariana Archipelago; the other utilize Amami, Okinawa, and the Philippines, migrating along the Ryukyu and Philippine Trench. The matching results also suggest that Hokkaido is possibly be utilized as a corridor between northern feeding areas and southern breeding areas at least by individuals migrating to Okinawa area.

Major and trace elements in the surface water of Tonle Sap Lake, Mekong River, and other tributary rivers in Cambodia.

To evaluate the seasonal water circulation of Tonle Sap Lake and its tributary rivers in Cambodia, the spatial distribution patterns of major and trace elements in surface water were investigated. Based on the similarity of the dissolved elemental concentrations, the water samples were mainly divided into the three groups: samples with relatively high percentages of Ca, Mo, and Sb (Subcluster B1); samples with high Si, Al, and Fe (B2); and samples with high Na, K, and Mg (B3). During the rainy season, the elemental composition of lake water (B1) appeared to be greatly influenced by the intrusion of water from the Mekong River (B1) through the Tonle Sap River (B1). During the dry season, the type of lake water shifted to B3, suggesting that the lake water stored during the rainy season was replaced by inflow from other tributaries and groundwater in its vicinity. Thus, the seasonal changes in the elemental composition of the lake water were largely controlled by surface water and groundwater circulation. The dissolved As concentration was higher in the lake water and during the dry season than that in the river water and during the rainy season, indicating the discharge of As from the lake's bottom sediment during the dry season. Although the redox cycling of Fe and Mn appeared to be less important due to the shallow water depth in the lake, there are potential risks of As poisoning induced by the formation of an anoxic water mass and increment in the concentration of phosphorus if eutrophication continues to progress.

Modulation of Cell Adhesion and Differentiation on Collagen Gels by the Addition of the Ovalbumin Secretory Signal Peptide.

Ovalbumin (OVA) is the most abundant protein in egg whites that is unnecessary in the egg yolk-based food industry. The development of OVA-based functional materials is of great interest. Collagen is a major component of the extracellular matrix. In this study, an OVA fragment, the OVA secretory signal peptide (OVA SP), was loaded in collagen gels, which were used as a cell scaffold for various types of cells. We examined the effect of OVA SP loaded in collagen gels to cell properties. The peptide was initially bound to the collagen fibers and then released from the gel. Our results indicate that the released OVA SP suppressed the integrin-mediated cell adhesion and focal adhesion formation. However, the adhesion of NIH3T3 cells was not suppressed by treatment with a chelating agent and an anti-ß1 antibody. Our results suggest that OVA SP nonspecifically interacts with cell surface proteins. The adhesion of various cell types on collagen gels were changed by the addition of OVA SP, depending on their integrin expression pattern. Additionally, the differentiation of MC3T3-E1 osteoblastic cells was promoted on the OVA SP-loaded collagen gels. This suggests that OVA SP may modulate both the differentiation and the adhesion of cells cultured on the collagen gels.

Low-temperature-modulated fruit ripening is independent of ethylene in 'Sanuki Gold' kiwifruit.

Fruit ripening in response to treatments with propylene, 1-methycyclopropene (1-MCP), and low temperature was characterized in 'Sanuki Gold' kiwifruit, Actinidia chinensis Planch. Propylene treatment immediately induced rapid fruit softening, increased AC-PG (polygalacturonase) and AC-EXP (expansin) mRNA accumulation, and stimulated an increase in the soluble solid concentration (SSC) and a decrease in titratable acidity (TA). After 3 d exposure to propylene, ethylene production and AC-PL (pectate lyase) mRNA accumulation were observed. 1-MCP treatment after 24 h exposure to propylene eliminated AC-PG mRNA accumulation and suppressed continued changes in SSC and TA. Application of 1-MCP at the start of the treatment, followed by continuous propylene exposure, markedly delayed fruit softening, and the expression of the cell wall-modifying genes, and changes in the SSC and TA, indicating that kiwifruit become insensitive to ethylene at least for 3 d following 1-MCP exposure. Surprisingly, significant fruit softening, mRNA accumulation of AC-PG, AC-PL, and AC-EXP, and decreased TA were observed without ethylene production in intact fruit stored at low temperature for 1 month, but not in fruit stored at room temperature. Repeated 1-MCP treatments (twice a week) failed to inhibit the changes that occurred in low temperature storage. These observations indicate that low temperature modulates the ripening of kiwifruit in an ethylene-independent manner, suggesting that kiwifruit ripening is inducible by either ethylene or low temperature signals.

The possible involvement of dopamine D3 receptors in the regulation of gastric emptying in rats.

AIM: The inhibitory effect of dopamine on gastric motility is thought to be mediated via a decrease in acetylcholine release resulting from stimulation of enteric neuronal dopamine D(2) receptors. The aim of this study was to investigate the possible involvement of the dopamine D(3) receptor in the regulation of gastric motility in rats using selective dopamine D(3) receptor agonists or a dopamine D(3) receptor antagonist. MAIN METHODS: Gastric emptying was assessed using the phenol red method after rats were treated with varying doses of dopamine D(3) receptor agonists or a dopamine D(3) receptor antagonist. KEY FINDINGS: S(+)-PD 128,907 (0.01-1 mg/kg, s.c.), a selective dopamine D(3) receptor agonist, dose-dependently delayed gastric emptying in rats. Other dopamine D(3) receptor agonists (i.e., R(+)-7-OH-DPAT [0.03-1 mg/kg, s.c.] and quinpirole [0.01-1 mg/kg, s.c.]) also delayed gastric emptying in rats. Both the selective dopamine D(1) and D(5) receptor agonist SKF-38393 and the selective dopamine D(4) receptor agonist PD 168,077 failed to delay gastric emptying in rats. The selective dopamine D(3) receptor antagonist (+)-S 14297 (10mg/kg, s.c.) partially inhibited the S(+)-PD 128,907-induced delay in gastric emptying. Although an administration of S(+)-PD 128,907 (1-100 µg/kg) into the 4th cerebral ventricle partially and dose-dependently delayed gastric emptying in rats, its administration into the lateral cerebral ventricle did not affect gastric emptying. SIGNIFICANCE: The results presented here suggest that peripheral dopamine D(2) receptors and, at least in part, dopamine D(3) and central dopamine D(2)/D(3) receptors play an important role in the regulation of gastric motility in rats.

Stereoselective synthesis of (E)-2-en-4-ynoic acids with ynolates: catalytic conversion to tetronic acids and 2-pyrones.

A highly torquoselective olefination of alkynoates to provide functionalized tetrasubstituted olefins, (E)-2-en-4-ynoic acids, is described. Addition of Brønsted acids dramatically switched the mode of the Ag(I)-catalyzed cyclization of the resulting enyne carboxylic acids to give either tetronic acids or 2-pyrones.

Effect of mosapride citrate hydrate on the colon cleansing action of polyethylene glycol electrolyte lavage solution (PEG-ELS) in guinea pigs.

Polyethylene glycol electrolyte lavage solution (PEG-ELS) is widely used for colon cleansing prior to colonoscopy and colonic surgery. It has recently been shown that coadministration of PEG-ELS and mosapride citrate hydrate (mosapride), a selective 5-HT(4)-receptor agonist, is clinically useful for barium enema examination as it allows adequate barium coating. However, there is no report showing that mosapride has beneficial effects on colon cleansing and its underlying mechanism in experimental animals. In the present study, we investigated the effects of mosapride on colonic transit and on the colon cleansing action of PEG-ELS in guinea pigs. Mosapride (10 - 20 mg/kg, i.g.) significantly enhanced colonic transit rate in guinea pigs. Although PEG-ELS alone showed adequate colon cleansing action, excess fluid remained in the colon. Coadministration of mosapride (20 mg/kg) and PEG-ELS, regardless of mosapride timing, reduced colonic content weight (dry residue and water amount) as compared to PEG-ELS alone. These findings suggest that mosapride enhances the colon cleansing action of PEG-ELS via an increase in colonic transit in guinea pigs, that is, it reduces not only fecal residue but also excessive fluid in the colonic lumen. It is therefore believed that coadministration of mosapride and PEG-ELS can allow better visualization in barium enema examination.

Pure rotational spectra of the CCCF radical.

Pure rotational transitions of a new carbon-chain radical CCCF in a supersonic jet have been observed for the first time using a Fourier-transform microwave spectrometer with a pulsed-discharge nozzle. The radical was produced by a pulsed electric discharge in a C(2)H(2) and CF(4) mixture diluted to 0.1% and 0.1% with Ne, respectively. Rotational transitions with spin and hyperfine splittings have been observed in the region from 9.1 GHz for N(K(a)K(c)) = 1(01)-0(00) to 27.3 GHz for N(K(a)K(c)) = 3(03)-2(02). The rotational constant, the spin-rotation interaction constant, and the hyperfine coupling constants due to the F nucleus have been precisely determined from the least-squares analysis, yielding B = 4555.8043(44), gamma(eff) = -7.105(16), b(F,eff) = 368(19), and c(eff) = -284.832(61) MHz. The determined molecular constants were compared with those obtained from high-level ab initio calculations and concluded that the CCCF radical has a bent ground state X(2)A(').

Fourier-transform microwave spectroscopy of the CCCCl radical.

Pure rotational spectra of the CCCCl radical in a supersonic jet have been observed for the first time by Fourier-transform microwave spectroscopy. The radical was produced by a pulsed electric discharge in a C(2)H(2) and CCl(4) mixture diluted to 0.3% and 0.2% with Ne, respectively. Transitions with spin and hyperfine splittings were observed for two isotopologs, CCC(35)Cl and CCC(37)Cl, in the region from 11.4 GHz for N=2-1 to 34.2 GHz for N=6-5. The molecular constants including the hyperfine coupling constants due to the Cl nucleus have been determined precisely. From the rotational analyses and high-level ab initio calculations, the molecular structure of the CCCCl radical is concluded to be bent in the ground electronic state.

The effect of alkynyl groups on torquoselectivity. Highly stereoselective olefination of alkynyl ketones with ynolates.

We have shown the first experimental evidence of the alkynyl substituent effect on torquoselectivity and succeeded in effecting the highly torquoselective olefination of alkynyl ketones to provide functionalized tetrasubstituted olefins.

Laser induced fluorescence spectroscopy of NC3O.

Laser induced fluorescence spectra of the NC(3)O radical in a supersonic jet have been observed. The radical was produced in a pulsed electric discharge of HC(3)N and O(2) diluted to 0.3% with Ar. A total of 17 vibronic bands with a radiative lifetime of approximately 30 ns have been observed in a region from 27 000 to 27 500 cm(-1). The observed vibronic bands are classified as (2)Pi(12)-(2)Pi(12), (2)Pi(32)-(2)Pi(32), and (2)Sigma-(2)Sigma types. The upper states of the (2)Sigma-(2)Sigma bands have large spin-rotation constants, which should be denoted as Sigma((+)) and Sigma((-)). From high-level ab initio calculations and rotational analyses, the observed transition was assigned to the B (2)Pi-X (2)A(") transition. Dispersed fluorescence spectra from the upper (2)Sigma and (2)Pi vibronic levels have also been observed, yielding fundamental vibrational frequencies for the nu(1), nu(2), nu(3), and nu(7) modes of the ground state.

Total synthesis of (+)- and (-)-sundiversifolide via intramolecular acylation and determination of the absolute configuration.

Intramolecular acylation of an organolithium leads to an efficient stereocontrolled total synthesis of both enantiomers of sundiversifolide. The absolute configuration was determined by HPLC analysis and allelopathy assay. The gamma-lactone moiety resulted from a butenolide was obtained by the condensation of a bicyclic alpha-hydroxyhemiacetal with Ph3P=CMe(CO2R).

Synthesis of multisubstituted furans, pyrroles, and thiophenes via ynolates.

An efficient synthetic method for the preparation of multisubstituted furans, thiophenes, and pyrroles using ynolates was developed. This novel formal [4 + 1] annulation by C2-C3 and C3-C4 bond formations includes cycloaddition, cyclization, decarboxylation, and dehydration as key steps.

Heteroatom-guided torquoselective olefination of alpha-oxy and alpha-amino ketones via ynolates.

Ynolates were found to react with alpha-alkoxy-, alpha-siloxy-, and alpha-aryloxyketones at room temperature to afford tetrasubstituted olefins with high Z selectivity. Since the geometrical selectivity was determined in the ring opening of the beta-lactone enolate intermediates, the torquoselectivity was controlled by the ethereal oxygen atoms. From experimental and theoretical studies, the high Z selectivity is induced by orbital and steric interactions rather than by chelation. In a similar manner, alpha-dialkylamino ketones provided olefins with excellent Z selectivity. These products can be easily converted into multisubstituted butenolides and gamma-butyrolactams in good yield.

[A case of stage I adenocarcinoma of the lung complicated with inflammatory lesions with an 8-year clinical course before surgery; CT and pathological correlations].

A nodular shadow near left hilum of lung was detected by chest radiography in November 1993 in a 79-year-old woman with hypertension and diabetes. Chest radiography in April 2001 revealed growth of the shadow. Chest CT showed a localized ground-glass opacity in the periphery of the lesion shadow. The resected specimen showed that the nodular shadow was papillary adenocarcinoma and the localized ground-glass opacity indicated the presence of inflammatory lesions. The pathological stage was stage I. Our case was a rare adenocarcinoma complicated with inflammatory changes including a granuloma that appeared 8 years later.

Stereoselective olefination of unfunctionalized ketones via ynolates.

Ynolates react with ketones at room temperature to afford alpha,beta,beta-trisubstituted acrylates (tetrasubstituted olefins) with 2:1-8:1 geometrical selectivities. This can be regarded as a new olefination reaction of ketones giving tetrasubstituted olefins in good yield, even in the case of sterically hindered substrates. The reaction mechanism involves cycloaddition of ynolates with a carbonyl group and subsequent thermal electrocyclic ring-opening of the resulting beta-lactone enolates. The stereoselectivity is determined in the ring-opening, which is regulated by torquoselectivity. In this paper, we describe the scope and limitations of olefination of ketones via ynolates and discuss the stereocontrol mechanism.

Effects of exercise and beta-blocker on blood pressure and baroreflexes in spontaneously hypertensive rats.

BACKGROUND: Exercise training or beta-blocker decreases high blood pressure (BP) and improves abnormal baroreflex function associated with hypertension. This study was undertaken to examine whether the effects of exercise training are additive to beta-blocker in spontaneously hypertensive rats (SHR). METHODS: At 5 weeks of age, SHR were allocated to four groups: sedentary control, exercise training, treatment with moderate dose of bisoprolol, and their combination. Systolic BP was monitored by the tail-cuff method under restrained conditions. Sigmoidal mean arterial pressure (MAP)-heart rate (HR) reflex curves were obtained in rats at 17 weeks of age under quiet conditions before and after atenolol to ensure sympathetic blockade and to determine the vagal component of gain. After studying baroreflex function, intrinsic HR was obtained by additional administration of atropine. RESULTS: Before atenolol, both exercise training alone and bisoprolol alone lowered resting MAP and HR, and decreased upper plateau (maximal tachycardia) and lower plateau (maximal bradycardia), resulting in decreased sympathetic component of HR range (upper plateau - intrinsic HR) and increased vagal component of HR range (intrinsic HR - lower plateau). After atenolol, both exercise training alone and bisoprolol alone increased the gain of vagal component. Exercise training had no additive effect on any parameters to bisoprolol except for systolic BP and HR measured by the tail-cuff method. CONCLUSIONS: Exercise training and bisoprolol have similar effects concerning resting hemodynamics and baroreflex function in SHR. Although additive effects of exercise training to bisoprolol are not evident under quiet, nonstressful conditions, some additive effects may be obtained under stress such as restrain.

Pneumonia in horses induced by intrapulmonary inoculation of Streptococcus equi subsp. zooepidemicus.

To evaluate the possibility that Streptococcus equi subsp. zooepidemicus (S.z) the causative bacterial agent of equine shipping fever pneumonia (ESFP), as well as to investigate its pathogenesis, 10 horses (seven Thoroughbreds and three Anglo-Arab species, ranging from 2-4 years in age) were experimentally inoculated, via an endoscope, into bronchus of the lung lobe with a dose of 30 ml of 1-7 x 10(8) CFU/ml of S.z. After inoculation, autopsy and pathological examinations were sequentially conducted 30 min, 1, 2, 3, 4, 17, 20 hr and 2 weeks later. Pneumonia induced by the intrapulmonary inoculation of S.z was characterized by small purulent pneumonic foci in the inoculated areas. With the lapse of time, these foci developed into serous hemorrhagic pneumonia, hemorrhagic purulent pneumonia, and then purulent, coagulation necrotic pneumonia. These pathomorphological characteristics of experimental pneumonia closely resemble those naturally occurring ESFP. There is strong evidence that S.z. is implicated as a causal factor in ESFP. S.z. grew in the mucus, exudate, and pulmonary effusions. Further, the bacteria showed resistance against phagocytosis by pulmonary alveolar macrophages (PAM) and neutrophils. Inhibition of PAM and neutrophil function is considered to be important in the development of pneumonia. With the progression of the disease, the neutrophils often adhered to the endothelial surface of the alveolar capillary lumen and played a role in generating coagulation necrosis of lung tissues.

Synthesis of alpha,alpha-dibromo esters as precursors of ynolates.

Aliphatic alpha,alpha-dibromo esters, precursors of ynolates, were synthesized via bromination of lithium alpha-bromo ester enolates with 1,2-dibromotetrafluoroethane in good yields. alpha-Trimethylsilyl-alpha,alpha-dibromo esters were synthesized via radical bromination.