Development and validation of a HPLC-MS/MS method for the analysis of fatty acids - in the form of FAME ammonium adducts - in human whole blood and erythrocytes to determine omega-3 index.

Recent scientific studies in the field of health and nutrition have unanimously affirmed the importance of consuming the omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), because of their cardioprotective properties. Fatty acid profiling in erythrocyte membranes allows the omega-3 index, which is a recognized indicator of the risk of developing cardiovascular disease, to be calculated. One consequence of the upward trend in healthy lifestyles and longevity is an increase in the number of studies into the omega-3 index, which requires a reliable method for the quantitative analysis of fatty acids. This article describes the development and validation of a sensitive and reproducible liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the quantitative analysis of 23 fatty acids (in the form of fatty acid methyl esters, FAMEs) in 40 µl of whole blood and erythrocytes. The list of acids includes saturated, omega-9 unsaturated, omega-6 unsaturated and omega-3 unsaturated fatty acids as well as their trans-isomers. The limit of quantitation was 250 ng ml-1 for C12:0, C16:0 and C18:0; and 62.5 ng ml-1 for other FAMEs, including EPA, DHA and trans-isomers of FAME C16:1, C18:1 and C18:2 n-6. Sample preparation for fatty acid (FA) esterification/methylation with boron trifluoride-methanol (BF3) has been optimized. Chromatographic separation has been carried out on a C8 column in gradient mode using a mixture of acetonitrile, isopropanol and water with the addition of 0.1% formic acid and 5 mM ammonium formate. As a result, the problem of separating the cis- and trans-isomers of FAME C16:1, C18:1 and C18:2 n-6 has been solved. The electrospray ionization mass spectrometry (ESI-MS) detection of FAMEs, in the form of ammonium adducts, has been optimized for the first time, which has made the method more sensitive that when the protonated species are used. This method has been applied to 12 samples from healthy subjects that consumed omega-3 supplements and has proven to be a reliable tool for determining the omega-3 index.

[One thousand endoscopic submucosal dissections. Experience of the national center]. / Odna tysyacha endoskopicheskikh podslizistykh dissektsii. Opyt natsional'nogo tsentra.

BACKGROUND: Endoscopic submucosal dissection (ESD) is a perspective method of organ-sparing treatment of benign colon tumors. MATERIAL AND METHODS: The study included 1.000 patients with colon neoplasms who underwent ESD between October 2016 and October 2021. All surgeries were performed under intravenous sedation. RESULTS: Mean dimension of tumors was 3.4 cm, median of surgery time - 51 (31; 101) minutes. Conversion of endoscopic approach occurred in 7.6% of cases. The main cause of conversion was unsatisfactory lifting in submucosal injection process. Incidence of en bloc and R0 resections was 84.1% and 68.3%, respectively. Postoperative morbidity was 2.9% that correlates with the world literature data. CONCLUSION: Endoscopic submucosal dissection is an effective and safe method for benign colon neoplasms. Considering high incidence of en bloc resection and low rate of local recurrence in benign neoplasms, further research of efficacy and safety of ESD in early colon cancer is needed.

[Endoscopic tattooing of colorectal neoplasms and laparoscopic surgery: technical aspects and recommendations]. / Éndoskopicheskii tatuazh obrazovanii tolstoi kishki i laparoskopicheskaia khirurgiia. Tekhnicheskie aspekty i rekomendatsii.

Accurate mapping of colorectal neoplasms is needed in many clinical situations. Intraoperative mapping of small lesions previously detected by endoscopy is often challenging, especially during laparoscopic surgery. Tumor location assessed during colonoscopy may be inaccurate because of limitations of the procedure. Small flat neoplasms with signs of invasiveness, which are hard to detect by palpation, hold a special place. The same situation is observed for nonradical endoscopic resection of malignant polyps or early cancer, when visual examination shows that the tumor masses have been completely resected but histological examination reveals the positive lateral or horizontal resection margin. Endoscopic tattooing is an effective, safe, and economically sound method to mark intraluminal colorectal neoplasms, which allows one to perform minimally invasive surgeries without using additional operating room resources.

NORMAL MEASUREMENTS OF PRESSURE IN ANAL CANAL DURING SPHINCTEROMETRY ON S4402 MSM AND WMP SOLAR GI DEVICES.

AIM: To assess normal measurements of pressure in anal canal during sphincterometry on S4402 MSM and WPM Solar GI devices. MATERIALS AND METHODS: The study included 126 patients with colonic polyps. inclusion criteria were absence of anal incon- tinence and defecation disorders. Seventy-three patients were assessed with S4402 MSM device, Included were 28 males (mean age 56,2±10,2 years) and 45 females (mean age 54,9±13,7 years). Fifty-three subjects were assessed via WPM Solar GI device: 23 women (mean age 51,4±11,1 years) and 30 males (mean age 65,1±15,9 years). RESULTS: Sphincterometry results using S4402 MSM device in males were as follows: mean resting pressure - 52,1+198 mm Hg; maximal resting pressure - 60,3±21,9mm Hg; mean pressure at voluntary contraction - 118,2±41,5 mm Hg and maximal pressure at voluntary contraction - 174,2±56,8 mm Hg. Corresponding values in females were 37,1±15,3 mm Hg, 43,8±15,5 mm Hg; 75,1±29,5 mm Hg and 99,1±39,7 mm Hg, respectively. Using WPM Solar GI sphincterometry the following figures were obtained in males: resting pressure - 43-61 mm Hg; maximal voluntary contraction pressure - 121-227 mm Hg; mean pressure - 106-190 mm Hg; maximal pressure with coughing test - 45-175 mm Hg; at straining minimal pressure decreased to 19-43 mm Hg (20-60%). In females the results were as follows: resting pressure - 41-63 mm Hg; maximal pressure at voluntary contraction 110-178 mm Hg; mean pressure - 88-146 mm Hg; maximal pressure at coughing test - 76-126 mm Hg, pressure decrease at straining to 28-52 mm Hg, relaxation up to 19-40%.

[Mutation analysis of K-ras protooncogene in colorectal adenocarcinomas and polyps in Russian patients].

To estimate diagnostic value of K-ras mutations during cancer risk group formation, they were studied in the samples of sporadic carcinomas (n = 33) and malignant (n = 13) polyps of large intestine obtained during surgery or polypectomy. Using PCR analysis, restriction analysis, SSCP analysis and automated sequencing, eight various point mutations were revealed. Six of them were located in codon 12 and two, in codon 13 of the K-ras gene. Mutation frequency in carcinomas, benign and malignant polyps was 43, 49, and 69%, respectively. In the healthy tissue of the large intestine, no changes in codons 12 and 13 in the K-ras gene were observed. Mutations in the groups of Russian patients examined partially overlapped. In patients with colorectal carcinoma the mutation frequency in the K-ras gene was not associated with disease onset age, location, and the extent of tumor differentiation while it was associated with the stage of tumor process. The maximum mutation frequency was revealed in polyps of patients over 70 years of age as well as in the adenomas of villous histology and large size ((1 cm). No correlation between the K-ras mutation frequency and the extent of polyp dysplasia was observed.

[Is subtotal resection of the colon actual for ulcerative colitis in the century of ileal reservoirs?].

The authors describe results of surgical treatment of 112 patients operated for a severe resistant form of ulcerative colitis (UC). In 26 (23.2%) of observations the operations were performed for complications, in 65 (58%)--for hormone-resistent, and in 21 (18.8%)--for hormone-dependent forms of UC. In 80 (71.4%) cases subtotal resection of the colon with ileo- and sigmostomies was fulfilled, in 26 (23.2%) a single stage ablation of all large intestine was fulfilled with the formation of permanent ileostomy and in 5 (5.4%) of observations the first step was finished with the formation of a reconstructive-plastic reservoir anastomosis. The number of complications in the nearest postoperative period was in 12.8% of patients, lethality was 2.3%. The development of a strict strategy of treatment for severe forms of the disease including the present-day using conservative and surgical methods of treatment depending on the individual character of each patient with UC made it possible to minimize the number of postoperative complications and decrease lethality and also to restore anal defecation with good functional results in 30.1% of operated patients.

[Results of ligature of distal branches of the upper rectal artery in chronic hemorrhoid with the assistance of Doppler ultrasonography]. / Rezul'taty ligirovaniia distal'nykh vetvei verkhnei priamokishechnoi arterii pod kontrolem ul'trazvukovoi doplerometrii pri khronocheskom gemorroe.

In October 2000 to February 2002 Doppler ultrasonography assisted suturing and ligation of hemorrhoidal arteries were performed in 102 patients with chronic hemorrhoids (57 men, 45 women, mean age 44 +/- 4.3 years). The mean duration of the disease was 10 +/- 4.6 years. Elimination of scarlet blood and hemorrhoidal prolapse without concomitant proctologic diseases were indications for surgery. Sixty-nine patients were followed up for more than 12 months, the follow-up lasted on the average 12 +/- 4 months. There were no clinical symptoms in 57 (82.6%) on the 69 patients. Combined ligation of arterial branches and nodes with latex rings was performed in 7 patients with stages III-IV hemorrhoids. Sclerotherapy was performed in 2 patients with stage III with occasional elimination of blood on defecation. Hemorrhoidectomy was performed in 3 (2.9%) patients with stage IV hemorrhoid due to ineffective ligation. Suturing and ligation of distal branches of the upper rectal artery decrease the inflow of arterial blood to hemorrhoidal nodes without venous outflow disorders. Fixation of hemorrhoidal nodes to the muscular wall eliminates the symptoms of hemorrhoidal prolapse. These factors lead to elimination of hemorrhoidal symptoms in 82.6% of the patients operated on.

Distribution of individual components of basement membrane in human colon polyps and adenocarcinomas as revealed by monoclonal antibodies.

Double-label immunofluorescence was used to monitor basement-membrane composition and integrity in 22 human colon polyps, 36 adenocarcinomas and 2 metastases. Cryostat sections were stained with polyclonal anti-laminin anti-serum combined with monoclonal antibodies (MAbs) to all major basement-membrane components (laminin, entactin/nidogen, collagen type IV and large heparan sulfate proteoglycan), as well as to keratin 8. In all adenocarcinomas, including mucinous, basement membranes were altered more at the invasive front than in the parenchyma. The degree of this alteration was inversely correlated with the level of tumor differentiation. An uncoordinated loss of basement membrane components (dissociation of markers), previously described by us in rat colon adenocarcinomas, was also found in human tumors. In the great majority of adenocarcinomas a pronounced stromal reaction was seen. It was manifested by the presence of fibrillar deposits of basement-membrane components, mainly of collagen type IV and/or heparan sulfate proteoglycan. This reaction was never observed in polyps and may be derived from myofibroblasts reported to accumulate in colon cancer stroma. The combined use of antibodies to basement-membrane components and to a specific keratin may constitute an adequate immunohistochemical test for the presence of invasion, and may be useful in the histologic analysis of polyps, especially in dubious cases.

[Results of endoscopic interventions in large and giant epithelial neoplasms of the large intestine]. / Rezul'taty éndoskopicheskikh vmeshatel'stv pri bol'shikh i gigantskikh épitelial'nykh novoobrazovaniiakh tolstoi kishki.

The paper presents the analysis of the results obtained for 319 patients at endoscopic interventions for large and giant tumors of the colon. Altogether 415 neoplasms were removed: 207 nodular, 126 creeping and 82 flat. The latter two were removed using specially developed technique of endoscopic electroresection of the intestinal mucosa along with the tumor. Malignancy was established in 16.1% of the cases. During the endoscopic procedure 2 patients were diagnosed to have colon perforations, 24 intensive hemorrhage which was stopped endoscopically in 22 cases. One patient developed subsequently necrosis of the intestinal wall. Surgical treatment was performed in 5 patients: for hemorrhage, colon wall perforation, malignancy in 2, 2 and 1 patient, respectively. One-eight-year follow-up of 316 patients reports cure in 217 (68.6%), relapses in 79 patients (71 of them were endoscopically successfully retreated), cancer in 15 and corrosive strictures in 5 patients.

[Complex evaluation of the healing of intestinal anastomoses in the early postoperative period]. / Kompleksnaia otsenka zazhivleniia kishechnykh anastomozov v rannem posleoperatsionnom periode.

Eighty-six patients were subjected to complex examination in the immediate postoperative period to check the condition of intestinal anastomoses. In 58 (67.4%) patients inflammation in the zone of the anastomosis was found to be negligible and produced no specific clinical manifestations; 28 (32.6%) patients had marked inflammatory changes in the region of the anastomosis. This condition, interpreted as anastomositis, is favourable for the development of incompetence of the sutures of the anastomosis, which was diagnosed in 4.7% of patients. Timely diagnosis of anastomositis and application of a complex of nonoperative measures is conducive to reduction of the frequency of postoperative complications and mortality.

[Healing of the anastomoses in restorative-reconstructive surgery of the large intestine]. / Zazhivlenie anastomozov v vosstanovitel'no-rekonstruktivnoi khirurgii tolstoi kishki.

An analysis of healing the intestinal anastomoses in 52 patients after restorative-reconstructive operations on the colon is presented. In 36.5% of the cases a complicated course of the early postoperative period was observed: anastomositis--in 28.8% and incompetence of the anastomosis sutures--in 7.7%. Such complications were mainly observed in patients older than 60 years of age with a concomitant pathology, with a one-trunk colostoma and when forming low colorectal anastomoses.

[Morphological and cytophotometric characteristics of polyps of the colon and rectum]. / Morfologicheskaia i tsitofotometricheskaia kharakteristika polipov tolstoi i priamoi kishok.

Material from 60 patients with colon and rectum polyps as well as "normal" mucous membrane (44 patients) taken at some distance from polyps are studied. DNA content was measured by a flow cytometry method (ICP-II) in parallel to morphological (cytological and histological) examination of biopsies. Analysis of material was performed by groups depending on the epithelial dysplasia degree in the adenomas. Diploid cells dominated in the adenomas. Significant increase of cells in S- and (G2 + M)-phases of cell cycles is noted in the adenomas with a severe epithelial dysplasia as compared to the adenomas with a mild or moderate dysplasia. Aneuploid cells are detected in 5 adenomas, 4 of them morphologically were those with a severe dysplasia including 2 cases in which it was difficult to differentiate between a severe dysplasia and carcinoma although the signs of invasion were absent.