Fourier transform infrared imaging and infrared fiber optic probe spectroscopy identify collagen type in connective tissues

Hyaline cartilage and mechanically inferior fibrocartilage consisting of mixed collagen types are frequently found together in repairing articular cartilage. The present study seeks to develop methodology to identify collagen type and other tissue components using Fourier transform infrared (FTIR) spectral evaluation of matrix composition in combination with multivariate analyses. FTIR spectra of the primary molecular components of repair cartilage, types I and II collagen, and aggrecan, were used to develop multivariate spectral models for discrimination of the matrix components of the tissues of interest. Infrared imaging data were collected from bovine bone, tendon, normal cartilage, meniscus and human repair cartilage tissues, and composition predicted using partial least squares analyses. Histology and immunohistochemistry results were used as standards for validation. Infrared fiber optic probe spectral data were also obtained from meniscus (a tissue with mixed collagen types) to evaluate the potential of this method for identification of collagen type in a minimally-invasive clinical application. Concentration profiles of the tissue components obtained from multivariate analysis were in excellent agreement with histology and immunohistochemistry results. Bone and tendon showed a uniform distribution of predominantly type I collagen through the tissue. Normal cartilage showed a distribution of type II collagen and proteoglycan similar to the known composition, while in repair cartilage, the spectral distribution of both types I and II collagen were similar to that observed via immunohistochemistry. Using the probe, the outer and inner regions of the meniscus were shown to be primarily composed of type I and II collagen, respectively, in accordance with immunohistochemistry data. In summary, multivariate analysis of infrared spectra can indeed be used to differentiate collagen type I and type II, even in the presence of proteoglycan, in connective tissues, using both imaging and fiber optic methodology. This has great potential for clinical in situ applications for monitoring tissue repair

Hypersegmentation of neutrophil nuclei in peripheral blood of patients with localized and advanced cancer of the larynx and laryngopharynx

Neutrophilic granulocytes have a wide spectrum of functional activity. In recent years, the functional significance of neutrophils in the development and course of malignant neoplasms has been discussed. It has been shown that neutrophilic granulocytes can play pro- or antitumor activity. The aim of the study was to assess the structural and functional features of neutrophils in patients with varying degrees of prevalence of cancer of the larynx and laryngopharynx. Forty-one patients (aged 35-67) with newly diagnosed cancer of the larynx and laryngopharynx were examined and divided into subgroups according to the TNM classification: the first subgroup (14 patients) with a localized tumor process consisted; and the second subgroup (27 patients) with a widespread tumor process. The relative and absolute number of neutrophils was assessed, and the neutrophil-lymphocyte ratio (NLR) was determined. The content of neutrophils with varying degrees of nuclear segmentation in the blood was calculated, the activity of myeloperoxidase, cationic proteins, alkaline phosphatase, and the degree of neutrophil activation in the NBT test was determined cytochemically. Concentration of interleukin-8 was determined using ELISA. In patients with cancer of the larynx and laryngopharynx the number of neutrophils (p = 0.045) and NLR (p = 0.033), as well as serum concentration of interleukin 8 (p = 0.011), increased compared to healthy individuals. The proportion of cells with hypersegmented nuclei in the neutrophil population (p < 0.001) and cytotoxic potential increased with the spread of tumor process. A direct correlation (r = 0.42, p = 0.03) was found between the T index, which reflects the volume of the tumor, and the content of hypersegmented neutrophils. It can be argued that such a simple and accessible laboratory parameter as the degree of segmentation of the nuclei of neutrophilic granulocytes can be used as one of the criteria to assess and predict the course of the tumor process

Effects of intranasal Imunofan administration upon phagocytic activity in treatment of exudative otitis media in children

Exudative otitis  media  in  childhood is most  often  associated with  chronic inflammation in the  nasopharyngeal area,  with  immediate participation of phagocytic cells. Our  paper  presents  the  data  on evaluation of clinical  and immunological efficacy of intranasal Imunofan use included into  complex therapy of exudative  otitis  media.  Dynamic observation (before  treatment, 1 and  3 months after treatment) of these parameters included regular  evaluation of the  neutrophil and  monocyte amounts in peripheral blood  and  in smear imprints from nasal mucosa, determination of myeloperoxidase activity in circulating neutrophils, and the content of interleukin IL-8 and IL-18  in the nasal washouts. The clinical status was assessed using a scoring system, which subjectively reflected the state of the nasopharynx and auditory function. Fourty-three children aged from 3 to 7 years with exudative  otitis media associated with chronic adenoiditis were examined. Patients of the first group (22 children) were treated using only conventional approaches (basic therapy). The patients from  the  second  group  (21 children) received  Imunofan in addition to the  basic therapy. The  control group consisted of 16 relatively  healthy  children. Before  treatment of the  children with exudative  otitis  media, an increase  in the relative content of monocytes in their blood,  a decreased activity of myeloperoxidase and lower concentration of IL-8  and  IL-18  in the  nasal  wash was observed  in comparison with  healthy  controls. No differences in severity  of clinical  symptoms were revealed  between  the  groups  of patients. Baseline  therapy was not  accompanied by positive  dynamics in the  clinical  pattern of the  disease.  Relative  monocytosis and reduced activity of neutrophilic myeloperoxidase persisted  in peripheral blood;  the concentration of IL-8  and IL-18  in the  nasal washings  remained low. Following intranasal use of Imunofan, the  number of circulating monocytes was restored by the  third  month from  the  start  of treatment, there  was an  increased activity  of myeloperoxidase registered  in blood neutrophils, as well as higher IL-8  and IL-18  concentrations in the nasal washings. Normalization of the phagocytos-related parameters, according to this scoring,  was associated with clinical remission of the disease. The revealed relationships between clinical data and the results obtained in the course  of laboratory research  suggest a positive effect of Imunofan as an agent that may enhance effectiveness of conventional basic therapy  of otitis media in children

Влияние гипертрофии больших слюнных желез на ультраструктуру сперматогенного эпителия крыс

Purpose. The aim of this study was to ascertain the characteristics of major salivary glands endocrine effect onspermatogenesis.Materials and methods. Mature white outbred male rats (2 months, 153 ± 18 g) consisted of the following groups (each containing 30 rats): intact, control, and group of rats subjected to multiple amputation of incisors. To achieve hypertrophy of major salivary glands multiple amputation of incisors was performed: incisors were cut to a level of 1-2 mm above the gingival margin under ether anesthesia once every 3 days within 2 weeks. Animals of the control group were anesthetized with ether at the same time. Rats were sacrificed by CO2 asphyxia after 2, 3, 4, 6, 8 and 10 weeks after the first amputation of incisors. Fragments of the rat testes were examined on a JEM-1400 “JEOL” (Japan) transmission electron microscope. On electron microscopy images the specific vacuolization of the cytoplasm of Sertoli cells, spermatogonia, spermatocytes and spermatids (standard units) was analyzed by the point counting method. In spermatogenic cells the proportion of mitochondria (%) with morphological signs of swelling was assessed.Results. Transient ultrastructural changes of Sertoli and spermatogenic cells develop in the rats convoluted seminiferous tubules as a result of multiple amputation of the incisors, such as phagosomes and pronounced vacuolization in the Sertoli cells cytoplasm, cytoplasm vacuolization and mitochondrial swelling in spermatogenic cells. Sporadic spermatogenic cells with signs of nuclear (chromatin fragmentation, its condensation on the periphery of the nucleus) and cytoplasm (destruction of membrane organelles) destruction appeared as a result of multiple incisors’ amputation. Ultrastructural changes of Sertoli and spermatogenic cells are most pronounced at 2-3 weeks, decrease at 4 week and are completely leveled by the 6th week of the experiment.Conclusion. Hypertrophy of major salivary glands, caused by multiple amputations of incisors, has similar to sialoadenectomy effect on the spermatogenic epithelium. Multiple incisors’ amputation cause transient  depression of granular convoluted cells function. Probably submandibular gland granular convoluted tubules cells endocrine factors make the greatest contribution to the regulation of spermatogenesis in rats. Цель. Выяснение особенностей эндокринного влияния больших слюнных желез на сперматогенез половозрелых крыс.Материалы и методы. Половозрелые белые беспородные самцы крыс (возраст 2 мес, масса тела (153 ± 18) г) составили три группы (по 30 особей): интактная, контрольная и крысы, подвергшиеся многократной ампутации резцов. Для оценки эндокринного  влияния эпителиоцитов ацинусов и протоков больших слюнных желез моделировали их гипертрофию путем многократной ампутации резцов. Крыс выводили из эксперимента на 2-, 3-, 4-, 6-, 8- и 10-ю нед после первой ампутации резцов. Семенники животных оценивали при помощи трансмиссионной электронной микроскопии. На электронограммах анализировали удельный объем вакуолизации цитоплазмы сустентоцитов, сперматогоний, сперматоцитов и сперматид (усл. ед.), в сперматогенных клетках оценивали количество митохондрий (%) с морфологическими признаками набухания.Результаты. В ранние сроки эксперимента в извитых семенных канальцах крыс развивается вакуолизация цитоплазмы сустентоцитов, а также ультраструктурные изменения сперматогенных клеток (вакуолизация цитоплазмы, альтерация митохондрий, разрушение мембранных органелл, фрагментация хроматина). Изменения структуры  сперматогенных клеток и сустентоцитов максимально выражен на 2–3-й нед, снижаются  на 4-й нед и полностью нивелируются к 6-й нед эксперимента.Заключение. Гипертрофия больших слюнных желез, вызванная многократной  ампутацией резцов, оказывает на сперматогенный эпителий влияние, схожее с эффектом сиалоаденэктомии. В результате многократной ампутации резцов угнетается  функциональное состояние клеток гранулярных извитых трубок поднижнечелюстных желез. Эндокринные факторы, которые вносят наибольший вклад в регуляцию  сперматогенеза у крыс, вырабатываются клетками гранулярных извитых трубок

Mineral maturity and crystallinity index are distinct characteristics of bone mineral

The purpose of this study was to test the hypothesis that mineral maturity and crystallinity index are two different characteristics of bone mineral. To this end, Fourier transform infrared microspectroscopy (FTIRM) was used. To test our hypothesis, synthetic apatites and human bone samples were used for the validation of the two parameters using FTIRM. Iliac crest samples from seven human controls and two with skeletal fluorosis were analyzed at the bone structural unit (BSU) level by FTIRM on sections 2–4 lm thick. Mineral maturity and crystallinity index were highly correlated in synthetic apatites but poorly correlated in normal human bone. In skeletal fluorosis, crystallinity index was increased and maturity decreased, supporting the fact of separate measurement of these two parameters. Moreover, results obtained in fluorosis suggested that mineral characteristics can be modified independently of bone remodeling. In conclusion, mineral maturity and crystallinity index are two different parameters measured separately by FTIRM and offering new perspectives to assess bone mineral traits in osteoporosis

ОСОБЕННОСТИ ВОСПАЛИТЕЛЬНОГО ИНФИЛЬТРАТА СЛИЗИСТОЙ ОБОЛОЧКИ ЖЕЛУДКА У ПАЦИЕНТОВ С ПРЕДОПУХОЛЕВЫМИ ИЗМЕНЕНИЯМИ И РАКОМ, АССОЦИИРОВАННЫМИ С ВИРУСОМ ЭПШТЕЙНА – БАРР

Characteristics of inflammatory infiltrate in the gastric mucosa of patients with gastric dysplasia (n=56) and gastric cancer (n=50) with different levels of humoral immune response to Epstein-Barr virus (EBV) and EBV viral load were studied. In patients with dysplasia of the gastric mucosa, the increase in antibody titers to VCA IgG leaded to a significant decrease in the level of lymphocytes, neutrophils and macrophages and an increase in the number of eosinophils and plasma cells. When the levels of IgA to viral capsid antigen (VCA) and IgG to EBV early antigens (EA) were increased, the number of neutrophils in the composition of the cellular infiltrate was significantly decreased. In gastric cancer patients with different levels of humoral immune response to EBV, the number of plasma cells and eosinophils in the inflammatory infiltrate of the tumor was decreased when increasing the titers of IgG to VCA and IgA to VCA. When VCA/IgA titer was high, the number of neutrophils in a tumor was decreased and the proportion of macrophages was slightly increased. The data obtained can serve as additional criteria for indentifying markers for viral infection of the gastric mucosa.Исследованы особенности воспалительного инфильтрата СОЖ у больных с предопухолевыми изменениями (n=56) и раком желудка (n=50) с различным уровнем гуморального иммунитета к вирусу Эпштейна – Барр (ВЭБ) и вирусной нагрузки ВЭБ. У больных с диспластическими изменениями СОЖ при увеличении титра антител к IgG к ВКА происходят изменения клеточного состава инфильтрата, выражающиеся в существенном снижении количества лимфоцитов, нейтрофилов и макрофагов и увеличении количества эозинофилов и плазмоцитов. При увеличении титра IgA к ВКА и IgG к РА ВЭБ в составе клеточного инфильтрата достоверно падает количество нейтрофилов. Состав клеточного инфильтрата опухоли у больных раком желудка с различным уровнем гуморального иммунитета к ВЭБ имел другие особенности: при увеличении титра IgG к ВКА и IgA к ВКА в воспалительном инфильтрате опухоли снижается количество плазмоцитов и эозинофилов. При высоких титрах IgA к ВКА снижается и количество нейтрофилов в опухоли и несколько возрастает доля макрофагов. Полученные данные могут служить дополнительными критериями для выявления маркеров вирусного поражения слизистой оболочки желудка

Ослабление местного мукозального иммунитета как фактор риска рецидивирования полипозного риносинусита

Assessment of the role of mucosal immunity in nasal polyposis recidivation was carried out. The density and the composition of inflammatory infiltrate in polyps from the patients with the primary and the recurrent nasal polyposis were researched. In nasal lavage the composition of the cells and the content of interleukin-4 and interferon-γ were estimated. It was shown that the polyposis recidivation occurs during the progression of local immune insufficiency.Проведена оценка роли местного иммунитета слизистой оболочки носа в рецидивировании полипозного риносинусита. В биоптатах полипов, удаленных у пациентов с первичным и рецидивирующим полипозом, исследовали плотность и состав воспалительного инфильтрата. В назальном лаваже оценивали клеточный состав и содержание интерлейкина-4 и интерферона-γ. Показано, что рецидивирование назальных полипов протекает на фоне прогрессирующей локальной иммунной недостаточности

Resveratrol Delays Age-Related Deterioration and Mimics Transcriptional Aspects of Dietary Restriction without Extending Life Span

22 páginas, 4 figuras.A small molecule that safely mimics the ability of dietary restriction (DR) to delay age-related diseases in laboratory animals is greatly sought after. We and others have shown that resveratrol mimics effects of DR in lower organisms. In mice, we find that resveratrol induces gene expression patterns in multiple tissues that parallel those induced by DR and every-other-day feeding. Moreover, resveratrol-fed elderly mice show a marked reduction in signs of aging, including reduced albuminuria, decreased inflammation, and apoptosis in the vascular endothelium, increased aortic elasticity, greater motor coordination, reduced cataract formation, and preserved bone mineral density. However, mice fed a standard diet did not live longer when treated with resveratrol beginning at 12 months of age. Our findings indicate that resveratrol treatment has a range of beneficial effects in mice but does not increase the longevity of ad libitum-fed animals when started midlife.This work was supported by grants from the American Heart Association (0425834T to J.A.B. and 0435140N to A.C.) and from the NIH (RO1GM068072, AG19972, and AG19719 to D.A.S.), (HL077256 to Z.U.), (HD034089 to L.W), (2RO1 EY011733 to N.S.W.), Spanish grant (BFU2005-03017 to P.N.), and by the generous support of Mr. Paul F. Glenn and The Paul F. Glenn Laboratories for the Biological Mechanisms of Aging.Peer reviewe

Молекулярные маркеры воспаления в бронхиальном содержимом при различных фенотипах тяжелой бронхиальной астмы

The complex immunologic estimation of inflammatory reactions in bronchial contents at 30 patients with various phenotypes of a serious bronchial asthma is spent. It is established that the population structure of lymphocytes and cytokines a profile of the induced phlegm reflects expression and character of inflammatory disturbances in respiratory tracts that will allow to use the given non-invasive method for revealing of molecular markers of serious forms of a bronchial asthma. At the serious terapevticheskirefractory form of a bronchial asthma «brittle» a phenotype in respiratory tracts total СD4+ -, СD19+ -cell and concentration interleukina-4 against depression interleukina-10 is enlarged. At the serious form of a bronchial asthma of a phenotype «the chronic asthma with the fixed bronchial obstruction» in a bronchial secret prevails CD8+ -lymphocytes, concentration interleukina-8, the transforming growth factor-β increases.Проведена комплексная иммунологическая оценка воспалительных реакций в бронхиальном содержимом у 30 пациентов с различными фенотипами тяжелой бронхиальной астмы (БА). Установлено, что популяционный состав лимфоцитов и цитокиновый профиль индуцированной мокроты отражает выраженность и характер воспалительных нарушений в дыхательных путях, что позволит использовать данную неинвазивную методику для выявления молекулярных маркеров тяжелых форм БА. При тяжелой терапевтически резистентной форме БА «brittle»-фенотипа в дыхательных путях увеличиваются общее количество СD4+ -, СD19+ -клеток и концентрация интерлейкина-4 на фоне снижения интерлейкина-10. При тяжелой форме БА фенотипа «хроническая астма с фиксированной бронхиальной обструкцией» в бронхиальном секрете преобладают CD8+ -лимфоциты, возрастает концентрация интерлейкина-8, трансформирующего фактора роста β