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Introduction: Several fluorescent proteins (FPs) and chromoproteins (CPs) are present in anthozoans and play possible roles in photoprotection. Coral tissues in massive corals often display discoloration accompanied by inflammation. Incidences of the pink pigmentation response (PPR) in massive Porites, described as inflammatory pink lesions of different shapes and sizes, has recently increased worldwide. FPs are reported to be present in PPR lesions, wherein a red fluorescent protein (RFP) appears to play a role in reducing reactive oxygen species. However, to date, the biochemical characterization and possible roles of the pigments involved are poorly understood. The present study aimed to identify and characterize the proteins responsible for pink discoloration in massive Porites colonies displaying PPRs, as well as to assess the differential distribution of pigments and the antioxidant properties of pigmented areas. Method: CPs were extracted from PPR lesions using gel-filtration chromatography and identified via genetic analysis using liquid chromatography-tandem mass spectrometry. The coexistence of CPs and RFP in coral tissues was assessed using microscopic observation. Photosynthetic antivity and hydrogen peroxide-scavenging activitiy were measured to assess coral stress conditions. Results: The present study revealed that the same CP (plut2.m8.16902.m1) isolated from massive Porites was present in both the pink spot and patch morphologies of the PPR. CPs were also found to coexist with RFP in coral tissues that manifested a PPR, with a differential distribution (coenosarc or tip of polyps' tentacles). High hydrogen peroxide-scavenging rates were found in tissues affected by PPR. Discussion and Conclusion: The coexistence of CPs and RFP suggests their possible differential role in coral immunity. CPs, which are specifically expressed in PPR lesions, may serve as an antioxidant in the affected coral tissue. Overall, this study provides new knowledge to our understanding of the role of CPs in coral immunity.
ABSTRACT
Absolute configuration at 12 stereocenters in the 36-membered macrocyclic ring portion of poecillastrin C (1) was disclosed by chemical degradation and NMR analyses of 1, chemical synthesis, and molecular modeling techniques.
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In the end of March 2018, an unprecedented food poisoning incident due to ingestion of the visceral balls of geoduck Panopea japonica occurred in Japan. The patient, presented with symptoms of numbness on the lips and general weakness, was diagnosed as paralytic shellfish poisoning (PSP). The patient immediately treated with the mechanical ventilation recovered and left the hospital after 3 days treatment. Saxitoxins (STXs) in the plasma and urinary samples collected from the patient on the first and second day after hospitalization were analyzed by ultra high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC/MS/MS) and liquid chromatography with post-column fluorescent detection (LC/FLD). The STXs levels of 499.1 and 6.0 µg/L of STX dihydrochloride equivalent (STX·2HCl eq.) were quantitated by LC/FLD in the urinary samples on the first and second day, respectively. In addition, geoducks harvested from the same areas of the PSP causative specimens after the incident were analyzed by LC/FLD, and the results showed the level of STXs in their whole bodies of the geoducks exceeding 0.8 mg STX·2HCl eq./kg which is the maximum levels of STX in CODEX STAN 292-2008. Prominent toxins in STXs that detected in urinary and geoduck samples and identified by UHPLC/MS/MS and LC/FLD were gonyautoxin-1+4 (GTX1+4). These results concluded that the incident was the food poisoning due to STXs accumulated in the geoducks. This is the first PSP case caused by consumption of geoducks in Japan. This is also the first PSP case that causative toxins are detected in urinary samples of patients involved in PSP in Japan.
Subject(s)
Saxitoxin , Shellfish Poisoning , Tandem Mass Spectrometry , Animals , Humans , Chromatography, High Pressure Liquid , JapanABSTRACT
Lipophilic marine biotoxin azaspiracids (AZAs) are produced by dinoflagellates Azadinium and Amphidoma. Recently, several strains of Azadinium poporum were isolated from Japanese coastal waters, and detailed toxin profiles of two strains (mdd421 and HM536) among them were clarified by several detection techniques on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and liquid chromatography-quadrupole time of flight mass spectrometry (LC-QTOFMS). In our present study, AZA analogues in seven strains of A. poporum from Japanese coastal waters (including two previously reported strains) were determined by these detection techniques. The dominant AZA in the seven strains was AZA2 accompanied by small amounts of several known AZAs and twelve new AZA analogues. Eight of the twelve new AZA analogues discovered in our present study were detected as bi-charged ions on the positive mode LC/MS/MS. This is the first report describing AZA analogues detected as bi-charged ions with hexose and sulfate groups in their structures.
Subject(s)
Dinoflagellida , Polyether Toxins , Spiro Compounds , Tandem Mass Spectrometry , Chromatography, Liquid , Japan , Dinoflagellida/chemistry , Marine Toxins/analysis , Spiro Compounds/analysisABSTRACT
An accurate and efficient method was developed for the determination of azaspiracid shellfish toxins (azaspiracids-1, -2, and -3), neurotoxic shellfish toxins (brevetoxins-2 and -3), diarrhetic shellfish toxins (okadaic acid and dinophysistoxins-1 and -2), and the amnesic shellfish toxin (domoic acid) in mussels (Mytilus galloprovincialis). Lipophilic marine biotoxins (azaspiracids, brevetoxins, and okadaic acid group) were extracted with 0.5 % acetic acid in methanol under heating at 60°C to improve the extraction efficiency of okadaic acid group toxins and then cleaned up with a C18 solid-phase extraction cartridge. Domoic acid was extracted with 50 % aqueous methanol and then cleaned up with a graphitized carbon solid-phase extraction cartridge. Lipophilic marine biotoxins and domoic acid were quantified by reversed-phase liquid chromatography coupled to electrospray ionization tandem mass spectrometry. The developed method had insignificant matrix effects for the nine analytes and good recoveries in the range of 79.0 % to 97.6 % at three spiking levels for all analytes except brevetoxin-2 (43.8-49.8 %). The developed method was further validated by analyzing mussel tissue certified reference materials, and good agreement was observed between certified and determined values.
Subject(s)
Bivalvia , Kainic Acid/analogs & derivatives , Oxocins , Polyether Toxins , Spiro Compounds , Tandem Mass Spectrometry , Animals , Okadaic Acid/analysis , Tandem Mass Spectrometry/methods , Chromatography, Reverse-Phase , Methanol , Chromatography, Liquid/methods , Shellfish/analysis , Marine Toxins/analysis , Bivalvia/chemistry , Solid Phase ExtractionABSTRACT
INTRODUCTION: Robot-assisted surgery is increasingly deployed in colorectal surgery, and decompression surgery using a stent is considered a standard treatment for malignant stenosis of the large intestine. Surgery after stent placement is also frequently performed. However, the anastomosis method remains controversial. PRESENTATION OF CASE: A 75-year-old woman visited our hospital's internal medicine department with chief complaints of bloody stool and constipation for the past year and colonoscopy was scheduled. After taking laxatives to prepare for treatment, abdominal pain was noticed and an emergency request was made. A diagnosis of colorectal malignant stricture and rectosigmoid junction cancer was made and a stent was placed during emergency colonoscopy. After intestinal decompression, a diagnosis of rectosigmoid junction cancer (UICC 8th; T3N0M0 Stage IIa) was rendered and robotic-assisted high anterior resection of the rectum and lymph node D3 dissection were performed. Reconstruction was performed using Gambee anastomosis outside the body cavity. The postoperative course was uneventful. DISCUSSION: The double stapling technique is simple, but in this case, the obstructed intestinal tract was swollen. Meanwhile, Gambee anastomosis, which allows adjustment of tightness, was considered effective. CONCLUSION: Gambee anastomosis is a valid option when robot-assisted rectal resection is performed after intestinal decompression with stent placement for malignant stricture of the rectosigmoid junction. It is important to select a hand-sewn or mechanical anastomosis by considering the condition of the organ to be anastomosed and the site of the anastomosis.
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A rare case of an ascending colon injury and ileal perforation in a 34-year-old male patient due to blunt abdominal trauma caused by a road traffic accident is reported in this study. This paper reports the clinical and imaging findings of seat belt syndrome. The seat belt syndrome primarily involves soft tissue injury; however, lacerations of the colon, small intestine, and mesentery have rarely been reported in the literature. However intestinal injuries, including bowel perforation and mesenteric injuries due to seat belt syndrome, must not be underestimated because they usually require emergency laparotomy because of accompanying peritonitis and hemorrhaging, and can be lethal if left untreated. Therefore, when an ascending mesocolon hematoma and free gas in the peritoneal cavity are present, gastrointestinal perforation due to seat belt syndrome should be suspected. In this case, gastrointestinal perforation was suspected based on the computed tomography findings, and emergency surgery was performed; the patient's course was uneventful without any postoperative complications. Early diagnosis and management are essential to prevent associated morbidity and mortality.
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α-Form hydrated crystals form a lamellar gel in which the alkyl chains of the amphiphilic molecules are hexagonally arranged within bilayers below the gel-liquid crystal phase transition temperature. In practice, the lamellar gel network with excess water is called an "α-gel", particularly in the cosmetics industry. In this study, the hydration or water sorption of amphiphilic materials in water vapor was assessed using a humidity-controlled quartz crystal microbalance with dissipation monitoring (QCM-D) technique. The amphiphilic materials used in this study were hexadecyl phosphate salts neutralized with L-arginine (C16P-Arg), CsOH (C16P-Cs), KOH (C16P-K), and NaOH (C16P-Na). Small- and wide-angle X-ray scattering measurements revealed that C16P-Arg and C16P-Cs yielded α-form hydrated crystals. Humidity-controlled QCM-D measurements demonstrated that C16P-Arg and C16P-Cs more readily underwent hydration or water sorption than C16P-K and C16P-Na. The key conclusion is that the significant hydration ability of C16P-Arg and C16P-Cs promotes the formation of the corresponding α-form hydrated crystals.