Relationship between GnRH-induced LH increase profiles in the serum and vaginal mucus of Japanese Black beef cows.

This study aimed to investigate the relationship between increases in the luteinizing hormone (LH) profiles in the serum and vaginal mucus of cows induced by gonadotropin-releasing hormone (GnRH). Samples for LH determination were collected from Japanese Black beef cows during estrus, which was induced with a controlled internal progesterone-releasing device and the administration of cloprostenol immediately before GnRH administration and every 30 min from the start of GnRH administration until 6.5 h. The peak serum LH concentration was clearly identified at 2.5 h post-GnRH administration, with serum concentrations returning to near-pre-GnRH-administration values after 6.5 h, whereas the peak vaginal mucus LH concentration was identified 4.5 h after GnRH administration. These results indicate that the LH secretion peak in vaginal mucus appeared about 2 h after peak LH secretion in the serum.

Pinpoint modification strategy for stabilization of single guide RNA.

The clustered regularly interspaced short palindromic repeats-CRISPR associated protein9 (CRISPR-Cas9) system, which includes a single guide RNA (sgRNA) and a Cas9 protein, is an emerging and promising gene editing technology that produces specific changes, including insertions, deletions, or substitutions, in desired targets. This approach can be applied in novel therapeutic areas for multiple cancers and genetic diseases, including Parkinson's disease, sickle cell disease, and muscular dystrophy. However, there are many limitations to its potential application to therapeutics. CRISPR-Cas9 activity without side effects, delivery of CRISPR-Cas9 to the target cell within the desired tissue including liver, lungs, brain and muscle and the expression of Cas9 endonuclease in the target cell are key factors in achieving therapeutic efficacy. Generally, single-stranded RNA is immediately degraded in cells and biological fluids such as serum, as chemically unmodified single-stranded RNA shows extremely poor stability against nuclease degradation. To overcome this limitation, sgRNA is chemically modified to obtain a highly stable sgRNA for efficient gene editing in cells and in vivo. Here, we identified the cleavage site of sgRNA for pinpoint modification in biological tissues using mass spectrometry and improved stability of pinpoint modified sgRNA in these fluids. Although improved efficiency provided by modified sgRNA has already been reported, we identified the cleavage site by mass spectrometry and revealed that the stability increased with the pinpoint modification strategy for the first time in this study. In future studies, the efficiency of pinpoint modification strategy for the potential application of sgRNA by systematic routes, including intravenous and subcutaneous administration will be assessed.

Biodegradation and Structural Modification of Humic Acids in a Compost Induced by Fertilization with Steelmaking Slag under Coastal Seawater, as Detected by TMAH-py-GC/MS, EEM and HPSEC Analyses.

The compost's humic acid (HA) content decreased when it was fertilized in coastal seawater with steelmaking slag, as confirmed. This study clarified the cause for this change by a detailed analysis of the structural changes in HAs based on the TMAH-py-GC/MS, HPSEC, and 3D-EEM spectra. An increase in the levels of pyrolysates of tannic acid with a low polymerization degree was attributed to the biodegradation of a high polymerized aromatic structure. Moreover, analyses of 3D-EEM, supported by HPSEC, indicated that approximately 20 kDa of the fluorescent matter was generated at the protein-like peaks (Ex/Em = 220/340 and 275/350 nm) in HAs derived from a mixture of compost with steelmaking slag. It would be caused due to the formation of HAs from the bacterial by-product by a catalytic reaction of the steelmaking slag. From these findings reported herein, we conclude that bio-degradation was a major reason for the decreased HA content, and the formation of HA from a part of the degradation products. This would be a reason for the structural modification of HA under the seawater condition.

Mechanism of the Elution of Iron from a Slag-Compost Fertilizer for Restoring Seaweed Beds in Coastal Areas-Characteristic Changes of Steelmaking Slag and Humic Acids Derived from the Fertilizer during the Elution Process.

The characteristic changes in steelmaking slag and humic acids (HAs) derived from a slag-compost fertilizer and their relation to the elution of Fe were evaluated in tank tests in Mashike, Hokkaido and Tsushima, Nagasaki. Analyses of iron, nitrogen and phosphate in the eluate, changes in the chemical states on the surface of the steelmaking slag, and the macro-structural features of the isolated HAs were investigated during the test. Temporal changes in Fe concentrations in the tanks were consistent with data collected in previous studies. Analyses of the surface by 57Fe Mössbauer spectroscopy showed that the concentration of Fe2+ in the fertilizer decreased and the ratio of Fe3+ increased, indicating that Fe2+ was preferentially eluted from the slag surface. The yields of HAs were significantly decreased during the test when steelmaking slag was mixed with compost. Changes in the UV-vis absorptivities indicated that de-aromatization had occurred. These results indicate that microorganisms that were present under these experimental conditions became activated by the mixing of compost with steelmaking slag, and are closely related to the continuous elution of Fe. The residual Fe in the fertilizer after the tests was investigated by analyzing seawater and the levels of extractable Fe. The trends for extractable Fe concentrations were consistent with the results obtained by 57Fe Mössbauer spectroscopy and UV-vis spectra.

Influence of Land Use on the Structural Feature of Sedimentary Humic Acids in Rivers in Northwest Hokkaido, Japan.

Colloidal organic matter is an important factor in our understanding of the transport of trace metals by rivers to coastal areas and the characteristics of organic matter in the sediment, including the effect of humic substances on the transport of such metals. The structural features of humic acids (HAs) derived from surface sediments collected from the mouths of four rivers in northwest Hokkaido, Japan, were evaluated from the perspective of land use in their basins. The sediments in two rivers (referred to as NS and SK) were lacking in HAs, but the sediments of the others (referred to as SM and PO) contained relatively high levels of HAs. UV-vis spectroscopic characteristics suggested that the HAs in SM and PO contained components derived from humified materials. The PO and SM basins contain rice paddy fields, and TMAH-py-GC/MS analysis showed that the pyrograms of paddy soil HA had a similar character to those of PO-HA and SM-HA samples. Thus, the differences of the structural features can be attributed to the land use in river basins. The findings indicate that the properties of organic river sediments are heavily influenced by the type of land use in the river basins.

Roles of Microbial Activity and Anthraquinone-2,7-disulfonate as a Model of Humic Substances in Leaching of Iron from Hematite into Seawater.

Fertilization with a mixture of steelmaking slag and compost can affect the supply of dissolved iron used to restore seaweed beds, however, the mechanisms of iron elution from the fertilizer are not well understood. In the present study, the microorganism was isolated from Fe-fertilizer incubated in coastal seawater for 6 months, and was identified as Exiguobacterium oxidotolerans by 16S rDNA sequencing. The iron elutability of the bacteria was proved based on the increasing of dissolved iron by incubation with Fe2O3 (hematite) under a seawater-like condition. The value of ORP was changed by inoculated of the bacteria from ca. 0 to ca. -400 mV, which is anticipated concerning to reduction of Fe. The concentration of eluted iron was largely depended on those of organic acids produced by bacteria. From the results, it was proved that E. oxidotolerans is capable of Fe reductive eluting of iron from Fe2O3 into seawater. Anthraquinone-2,7-disulfonate (AQDS), which can play as an electron acceptor/donor between microbe and insoluble Fe2O3 particles, enhanced the effect of iron bio-leaching.

Design, Synthesis, and Evaluation of the Highly Selective and Potent G-Protein-Coupled Receptor Kinase 2 (GRK2) Inhibitor for the Potential Treatment of Heart Failure.

A novel class of therapeutic drug candidates for heart failure, highly potent and selective GRK2 inhibitors, exhibit potentiation of ß-adrenergic signaling in vitro studies. Hydrazone derivative 5 and 1,2,4-triazole derivative 24a were identified as hit compounds by HTS. New scaffold generation and SAR studies of all parts resulted in a 4-methyl-1,2,4-triazole derivative with an N-benzylcarboxamide moiety with highly potent activity toward GRK2 and selectivity over other kinases. In terms of subtype selectivity, these compounds showed enough selectivity against GRK1, 5, 6, and 7 with almost equipotent inhibition to GRK3. Our medicinal chemistry efforts led to the discovery of 115h (GRK2 IC50 = 18 nM), which was obtained the cocrystal structure with human GRK2 and an inhibitor of GRK2 that potentiates ß-adrenergic receptor (ßAR)-mediated cAMP accumulation and prevents internalization of ßARs in ß2AR-expressing HEK293 cells treated with isoproterenol. Therefore, 115h appears to be a novel class of therapeutic for heart failure treatment.

Primary closure of a spontaneous esophageal rupture under hand-assisted laparoscopy: a case report.

Spontaneous rupture of the esophagus, which is also known as Boerhaave's syndrome, is a rare life-threatening condition that requires urgent surgical management. The optimal treatment involves surgical repair of the esophageal defect, which is usually accomplished via laparotomy, thoracotomy, or both, and mediastinal debridement. Here, we report a case of spontaneous rupture of the esophagus that was treated with suturing repair and drain insertion using a hand-assisted laparoscopic approach.

[A Case of Interstitial Pneumonitis Induced by Lapatinib plus Letrozole].

Lapatinib is an orally bioavailable dual inhibitor of the intracellular domain of both the HER2 protein and the epidermal growth factor receptor. This dual inhibitor can effectively prevent the downstream signal transduction initiated by a tyrosine kinase, thereby reducing the proliferation rate of tumor cells. Lapatinib was demonstrated to be beneficial in patients with HER2-positive locally advanced and metastatic breast cancer. We present here a case of interstitial pneumonitis that occurred after lapatinib treatment. A 79-year-old woman was diagnosed with Stage III infiltrating ductal carcinoma of the right breast. She underwent a modified radical mastectomy in January 2009, followed by anthracycline and paclitaxel plus trastuzumab administration. In November 2015, lung metastatic disease was detected. Therefore, lapatinib plus letrozole administration was initiated. Twelve days after starting treatment, she developed severe eruptions along with dyspnea. Radiography and a CT scan showed a diffuse ground glass shadow. Both her symptoms and the radiographic findings improved dramatically after the start of high-dose corticosteroid therapy. Clinicians should be aware that lapatinib has the potential to cause lung injury.

[Local Microwave Hyperthermia for Advanced or Recurrent Breast Cancer].

The purpose of our study was to test the efficacy and toxicity of hyperthermia for treating breast cancer. Ten patients received treatment (AC, paclitaxel, S-1, and aromatase inhibitor) in combination with hyperthermia. The hyperthermia device was a microwave heating device with water loaded and water-cooled waveguides. The temperature was monitored subcutaneously in the skin under the aperture of the waveguide. Two patients had a partial response to treatment with only mild toxicity (grade 1 acute skin toxicity). Therefore, hyperthermia combined with chemotherapy for treating breast cancer seems to be effective and generally tolerable. A larger patient cohort is needed to confirm these results in the future.

A case of deep femoral artery aneurysm.

A 58-year-old man presented with paralysis and pain in the left leg, and a mass was found in his thigh. Because of the growth of the mass and the worsening of his symptoms, the patient visited our hospital. Multidetector computed tomography revealed a large deep femoral arterial (DFA) aneurysm. Surgical intervention was planned because of the large size of the aneurysm, the high risk of perforation and the worsening symptoms. Aneurysmectomy and revascularization of the distal DFA with an artificial blood vessel graft were performed. DFA aneurysms are extremely rare. These aneurysms have a high rate of rupture, and surgery plays an important role in their treatment. However, standard methods have not yet been established because of their rarity of DFA aneurysm. We describe a case of DFA aneurysm in a patient who was successfully treated with aneurysmectomy and revascularization with an artificial blood vessel graft.

[The synergistic effect of local microwave hyperthermia and chemotherapy for advanced or recurrent breast cancer].

The purpose of our study was to test the efficacy and toxicity of hyperthermia in conjunction with chemotherapy for breast cancer. Between 2009 and 2014, 6 patients diagnosed with breast carcinoma were selected for this retrospective study. Patients received standard chemotherapy (AC followed by paclitaxel ) in combination with hyperthermia. The hyperthermia device employed microwave heating with water loaded and water-cooled waveguides. The temperature was monitored subcutaneously in the skin under the aperture of the waveguide. Following hyperthermia therapy, 4 patients had a partial response to treatment and the toxicity was mild, consisting of Grade 1 acute skin toxicity. Therefore, hyperthermia combined with chemotherapy for breast cancer seems to be effective and generally tolerable. A larger patient cohort is needed to confirm these results.

Determination of labile Fe(II) species complexed with seawater extractable organic matter under seawater conditions based on the kinetics of ligand-exchange reactions with ferrozine.

A fertilizer, comprised of a mixture of steel slag and compost, was used to restore seaweed beds in barren coastal areas. Complex Fe(II) species, supplied by steel slag, play a significant role in supplying Fe(II) to coastal areas and stimulating seaweed growth. Seawater extractable organic matter (SWEOM) from compost is generally assumed to serve as a chelator of Fe(II) in the fertilizer. It is considered that the bioavailability of Fe(II)-SWEOM complexes is higher in the dissociable (labile) species. In the present study, a method for determining labile species of Fe(II)-SWEOM complexes in seawater (pH 8.0, I = 0.7) was developed. The method is based on a ligand-exchange reaction between SWEOM and ferrozine (FZ). Because Fe(II) is readily oxidized to Fe(III) under normal seawater conditions, ascorbic acid was added as an antioxidant. The coloring for the Fe-FZ complex in the presence of SWEOM was retarded. This retarding can be attributed to a ligand-exchange reaction between FZ and labile Fe(II)-SWEOM complexes. Conditional binding constants for the labile Fe(II)-SWEOM complexes and binding capacities of labile sites in SWEOM to Fe(II) were evaluated for a variety of total Fe(II) concentrations.

The utility of hyperthermia for local recurrence of breast cancer.

BACKGROUND: Hyperthermia has long been used in combination with chemotherapy or radiation therapy for the treatment of superficial malignancies, in part due to its sensitizing capabilities. Patients who suffer from superficial recurrences of breast cancer have poor clinical outcomes. Skin metastases may particularly impair the quality of life due to the physical appearance, odor and bleeding. CASE PRESENTATION: A 66-year-old woman underwent mastectomy and axillary lymph node dissection for breast cancer. Nine years post-operatively, local metastases developed in the left axillary area (measuring 5 cm in diameter). Initially the tumor did not respond to radiation therapy and chemotherapy. Therefore, we added hyperthermia combined with them. Eight weeks later, the tumor became nearly flat and the patient noted improved activity in her daily life. CONCLUSION: Hyperthermia may accelerate the antitumor effects of radiation therapy and chemotherapy. This treatment provides an alternative for unresectable breast cancer skin metastases.

Binding characteristics and dissociation kinetics for iron(II) complexes with seawater extractable organic matter and humic substances in a compost.

A steel-slag/compost fertilizer can be useful in supplying complex Fe(II) species to barren coastal regions. Seawater extractable organic matter (SWEOM) was examined for use as a novel chelator of Fe(II) in the compost. The dissociation kinetics for Fe(II)-SWEOM were evaluated, based on the rate of ligand-exchange with ortho-phenanthroline. The ΔH(‡) for the Fe(II)-SWEOM (19 kJ mol(-1)) was significantly smaller than the corresponding values for Fe(II) complexes with humic substances (27 kJ mol(-1)), suggesting that the Fe(II)-SWEOM is kinetically less stable.

Simultaneous development of adenocarcinoma and gastrointestinal stromal tumor (GIST) in the stomach: case report.

BACKGROUND: Gastrointestinal stromal tumors (GISTs) and adenocarcinoma are distinct neoplasms originating from different cell layers. Approximately 20% of patients with GIST develop other cancers. CASE PRESENTATION: We report a case of the coexistence of adenocarcinoma and gastrointestinal stromal tumor (GIST). Gastric endoscopy showed the ulcerated tumor with bleeding along the lesser curvature of the proximal stomach and a submucosal nodule that measured about 3 cm in diameter in the lower part of the stomach body. Their pathological examination showed gastric cancer (poorly differentiated diffuse adenocarcinoma) and GIST (low-risk category). Further, immunohistochemical staining for C-kit and CD34 was positive, while that for SMA and S-100 was negative. CONCLUSION: Although it is not easy to speculate on the coexistence of adenocarcinoma and GIST, pre-and post-operative diagnoses may be essential, and such cancer development is not considered to be unusual.

Design, synthesis, and structure-activity relationships of spirolactones bearing 2-ureidobenzothiophene as acetyl-CoA carboxylases inhibitors.

The co-crystal structure of the human acetyl-coenzyme A 2 (ACC2) carboxyl transferase domain and the reported compound CP-640186 (1b) suggested that two carbonyl groups are essential for potent ACC2 inhibition. By focusing on enhancing the interactions between the two carbonyl groups and the amino acid residues Gly(2162) and Glu(2230), we used ligand- and structure-based drug design to discover spirolactones bearing a 2-ureidobenzothiophene moiety.

[A successful case treated with S-1 and zoledronic acid for multiple bone metastasis of breast cancer].

The patient was a 55-year-old woman undergoing surgery for T2, N1, M0, stage II B breast cancer in November 1997. UFT and tamoxifen were administered as adjuvant chemotherapy. S-1 and zoledronic acid were then administered for the recurrence of multiple bone metastasis detected in January 2008. A significant improvement was achieved, and a long-term positive effect was confirmed until an exacerbation of bone metastasis in February 2010. S-1 and zoledronic acid therapy have few adverse drug reactions, they promptly manifest their effect, and are favored as a first choice for treating bone metastasis of breast cancer.

[A long-term survivor treated with S-1 and gemcitabine for recurrence following an operation for advanced gall bladder cancer].

The patient was a 75-year-old woman who had undergone an operation for T4, N0: stage IV a gall bladder cancer in May of 2003. UFT was given as adjuvant chemotherapy. Then S-1 and gemcitabine (GEM) were given because of the recurrence of peritoneal dissemination in June 2006. Though a partial response was maintained, it was judged as progressive disease in July 2007. The peritoneal tumor was solitary, and an operation was performed. S-1 and GEM were given again in January 2008 because of multiple lung metastases. Long-term survival was confirmed until she died of peritonitis carcinomatosa in August 2009. To our knowledge, no case of a long survivor with S-1 and GEM for recurrence after a gall bladder cancer operation has been reported in the literature. Thus, future clinical trials and the accumulation of further cases are warranted.

Photoelectron spectroscopy and Ab initio calculations of peroxy form of SO(4)(-) anion.

A photoelectron spectrum is reported for gas-phase SO(4)(-) formed in an electron-impact ionized free jet of SO(2)(1%)/O(2)(10%)/Ar. The vertical detachment energy (VDE) of the SO(4)(-) species is determined to be 3.78 +/- 0.02 eV, which is significantly smaller than the VDE value reported for the sulfur-centered form of SO(4)(-) extracted from an Na(2)S(2)O(8) solution. With the aid of MP2/6-311+G(2df) calculations, the newly formed species is identified as a peroxy form of SO(4)(-), which is well described as OOSO(2)(-)(C(1), (2)A) having an S-O linkage between the O(2) and SO(2) units. The electronic properties of OOSO(2)(-) are characterized by the singly occupied molecular orbital (SOMO) constructed by an out-of-phase superposition of the 1a(2) orbital of SO(2) and the half-filled 2ppi(g)* of O(2)(-). On the basis of these findings, a possible formation mechanism of OOSO(2)(-) is discussed in terms of a "solvent-mediated" addition reaction, where O(2)(-) stabilized by solvation attacks SO(2) while retaining the 2ppi(g)* excess electron.