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PubMed1999

Application of capillary nongel sieving electrophoresis for gene analysis.

Capillary electrophoresis (CE) has proved to be a strong tool for DNA analysis and has found abundant applications in the fields of restriction fragment sizing, mutation screening, polymerase chain reaction (PCR) product characterizing and forensic identifying. CE may be the main alternative to slab gel electrophoresis. Capillary nongel electrophoresis is the most favorable mode when aiming for this purpose because of its advantages of long lifetime, easy operation, good reproducibility, and low expense. In this paper, a new kind of sieving matrix, with mannitol as the additive for capillary electrophoresis, as well as related methods and their application for gene analysis were reported. Nine DNA fragments amplified by multiplex PCR from a normal dystrophin gene were well separated by this system. Three different deletions were found in Duchenne muscular dystrophy (DMD) patients. Three to four copies of the sex-determination region of the Y chromosome (SRY) gene, as well as the phenylalanine hydroxylase (PAH) gene, could be detected in mixed samples. The frequencies of short tandem repeats (STR) in PAH genes was analyzed in 61 normal Chinese individuals and 6 phenylketonuria families. One case of prenatal gene diagnosis was performed. By using this matrix, CE coupled with reverse transcription PCR (RT-PCR), the analysis of the alternative splicing expression pattern of the fragile X mental retardation 1 (FMR1) gene in adult lung tissue was achieved.

Adult↗
PubMedDate not supplied

[Metabolic functions of the skin].

20 percent of the skin's energy expenditure goes on metabolic exchanges. It is in the microcirculation that the metabolic flow can be identified. The pre-terminal structures responsible for the variability of the outflow must be distinguished from the terminal structures where the hydraulic translations take place. In the network stereo-composed of terminal capillaries may be seen incidental flux (from the arteriole to the venule), interrupted transitory flux, and inverted flux. These circulatory mechanisms are admirably adapted to the variable and rhythmic needs of the metabolism. But the whole mechanism is fragile.

Biomechanical Phenomena↗
PubMed2003

Effects of high hydrostatic pressure on several sensitive therapeutic molecules and a soft nanodispersed drug delivery system.

PURPOSE: According to the development in the last decade of industrial processes using high hydrostatic pressure (HHP) for preservation of several commercial food products, novel sterilization or decontamination processes for pharmaceutical products could be conceivable. The aim of this work is to evaluate the effects of HHP on the integrity of insulin and heparin solutions, suspension of monoclonal antibodies and Spherulites. METHODS: High performance liquid chromatography, thin layer chromatography, capillary electrophoresis assays, ELISA tests, laser granulometry and spectrophotometry analyses have been performed to compare HHP treated drugs (in a domain of pressure and temperature ranging respectively from 20 up to 500 MPa and from 20 degrees C up to 37 degrees C) vs. untreated ones. RESULTS: No difference has been detected except for monoclonal antibodies that are altered above 500 MPa. CONCLUSIONS: The structure integrity of sensitive molecule due to the small energy involved by HHP and the development of industrial plants (intended for the decontamination of food products) confer to this technology the potential of a new method for sterilization of fragile drugs and an original alternative to aseptic processes and sterilizing filtration.

Antibodies, Monoclonal↗
PubMed1989

[Biomechanics of the bones and skeleton. III. Microstructure].

The authors analyzed micro- and ultrastructure of Haversian system--osteon--from the aspect of mechanics--biomechanics. The evaluation takes into account biological factors and instead of mathematical formulae and calculations prefer a comparison with technical constructions, particularly for emphasizing differences or similarities. The analysis of osteon revealed that it consists of fibres, net and tubes. The orientation of tube-like lamellae is in the direction of compressive strain, whereas the resultant course of torison and bending stress may be well observed in the direction of collagen fibres. A slight assymetry of osteons suggests that they are stressed by pressure in a slightly excentric direction. In the course of stress lengthwise there is the development of extension of the Haversian canal supporting the blood flow in the capillary. In the course of stress in the osteon lengthwise there are developing pressure, torsion and bending strains. Liquid phase can, together with collagen, absorb considerable portion of kinetic energy, thereby decreasing the strain of external strengths to act on the fragile mineralized component. Without the viscous and collagen elements the mineralized component would not be able to resist the dynamic forces which develop during the stresses in the bone tissue. Identification of mechanic phenomena of osteon provides information on structural principle of the whole bones. The authors applied in their study the knowledge from the discovery of USSR No. 181 and a discovery of CSSR No. 43.

Biomechanical Phenomena↗
PubMed2000

Use of capillary electrophoresis for high throughput screening in biomedical applications. A minireview.

Diagnosis of inherited diseases or cancer predispositions frequently involves determination of specific mutations or polymorphisms. The number of characterized monogenetic and polygenetic diseases is significantly rising every year. As a result, an increasing number of patient samples with a rising complexity of genetic diseases require molecular diagnostics. In order to apply genetic analyses to large groups of patients or population screening, automation of a sensitive and precise method is highly desirable. Capillary electrophoresis (CE) facilitates the development of methods which can rapidly process large number of patient samples in an automated fashion. In contrast, conventional techniques including Southern blotting, sequencing or standard gel electrophoresis are time consuming, cost ineffective and require substantial amounts of each specimen. Robustness, ease of operation, good reproducibility and low cost are the main advantages of CE. Currently, most protocols adapted to automated CE represent (i) analyses of DNA fragment length or DNA restriction patterns (RFLP), (ii) analyses of single-strand conformation polymorphism (SSCP) and (iii) microsatellite analyses. Recently, automated detection of variations in the FRAXA (CGG)n region (fragile X syndrome), LDL receptor gene, p53 gene, MTHFR (methylenetetrahydrofolate reductase) gene, HFE gene and others has been established on CE systems. These applications clearly demonstrate the suitability of CE for high throughput screening in medical applications.

Clinical Laboratory Techniques↗
PubMedDate not supplied

[Ecchymotic patches of the fingers and Gardner-Diamond vascular purpura].

Ecchymotic patches on the fingers and vascular purpura in Gardner-Diamond syndrome are two benign but recurrent clinical disorders occurring chiefly in young women. They involve superficial cutaneous hemorrhagic signs. Both disorders testify to microvascular fragility without perturbation of general hemostasis. Diagnosis is easy for those familiar with the disorders. Ecchymotic episodes accompanied by sharp pain begin at the roots of the fingers. Microtraumatism is soon seen to be involved. Ecchymosis develops at the flexor muscles of the fingers but "capillary" resistance is only reduced in one third of cases. The pathogenic explanation involves rupture of a small post-capillary vein due to disorders in the control system of kinins and local metabolites. The autoerythrocyte sensitization syndrome described in 1955 by Gardner and Diamond is characterized by painful and febrile episodes followed by purpuric and ecchymotic lesions of the skin on various parts of the body, but not necessarily on the legs. Dysneurotonic effects are frequent but diagnosis is based upon promotion of the disorder by intradermal or subcutaneous injection of a minimal quantity of autologous blood. Immunological effects are considered but the ailment involves microvascular control disorders, notably of the capillary-vein segment, in particular at Copley's endo-endothelial fibrin film. Imbalance in fibrin formation and lysis is associated with perturbation of the kinin and serotonin systems. In addition to the use of certain bioflavonoids and calcium inhibitors, prevention and treatment should involve consideration of drugs which affect plasticity in the vascular wall and pericapillary interstitial conjunctiva and the rheological properties of blood flow and interstitial tissue.

Ecchymosis↗
PubMed1991

Enhancement of metastatic potential by gamma-interferon.

Preincubation of murine colon 26 colon adenocarcinoma cells with gamma-interferon (IFN-gamma), but not alpha-interferon, produced a significant increase in experimental pulmonary metastases in syngeneic BALB/c and T-cell-deficient BALB/c nude mice. The enhancement was seen after as little as 1 h of exposure to 1 unit/ml of IFN-gamma and persisted for at least 72 h following removal of the cytokine. IFN-gamma exerted its effects by increasing the pulmonary retention of cells during the first 6 h following tumor cell injection. During this period all cells visualized in the lung were trapped in pulmonary capillaries. The enhancement was not due to modulations in class I major histocompatibility complex surface antigen expression; nor was it due to alterations in cell size, adhesion to components of the extracellular matrix in vitro, heterotypic or homotypic adhesion, sensitivity to lysis by activated peritoneal macrophages, osmotic fragility, enhancement of surface class II major histocompatibility complex antigen expression, or enhancement of intercellular adhesion molecule-1 (ICAM-1). Colon 26 was completely resistant to natural killer cell-mediated lysis in vitro, and IFN-gamma did not modulate the ability of colon 26 to form conjugates with isolated splenocytes. In vivo elimination of anti-asialo GM1 + cells increased pulmonary metastasis, and in such mice, there was no longer a difference in metastatic potential between control and IFN-gamma-treated cells. We conclude that low doses of IFN-gamma generated at the site of the tumor by host-infiltrating cells or during cytokine therapy could enhance the survival of tumor cells in the circulation and enhance their metastatic potential.

Adenocarcinoma↗
PubMed1994

Moyamoya disease: the disorder and surgical treatment.

OBJECTIVE: To discuss the clinical features of moyamoya disease, the studies that aid in diagnosing this disorder, and the reported outcomes of surgical treatment. DESIGN: We review the manifestations of moyamoya disease in children and adults and the recent reports of the various surgical procedures. MATERIAL AND METHODS: Moyamoya disease is a chronic cerebrovascular disorder in which stenosis of the major arteries of the circle of Willis at the base of the skull progresses to occlusion. The diagnosis is based on the angiographic findings of the "puff of smoke" appearance of the abnormal capillary vessels at the base of the skull. Three surgical procedures are used to manage this disease: anastomosis of the superficial temporal artery to the middle cerebral artery, encephalomyosynangiosis, and encephaloduro-arteriosynangiosis. RESULTS: In children with this disease, cerebral ischemic events, including strokes, occur. In adults, the fragile abnormal vessels can rupture and cause intracerebral hemorrhage. The mortality rate for adults is higher than that for children. Most published reports support the efficacy of surgical treatment in children but not in adults. CONCLUSIONS: The natural history of moyamoya disease is poor; neurologic deterioration due to strokes and hemorrhage is progressive. Seizures and intellectual deterioration can occur.

Adolescent↗
PubMed2026

Clinical Application of Long-Read Sequencing for FMR1 Gene Mutation Detection in Populations From Shandong, China.

BACKGROUND: Fragile X syndrome (FXS) is a common inherited intellectual disability. In this study, long-read sequencing was used for the FMR1 gene detection. METHODS: Men with familial inherited intellectual disability and women with indications for FXS screening were defined as high-risk populations and were included in this study along with non-high-risk reproductive-aged women. PCR-capillary electrophoresis was used for preliminary screening of non-high-risk reproductive-aged women, and long-read sequencing was performed on abnormal samples and samples from high-risk populations. Prenatal diagnosis using long-read sequencing was performed for pregnant women in need. RESULTS: The prevalence of mutation in high-risk females was 3.10% (7/226). 3 mutations were detected in male samples, with a mutation ratio of approximately 8.3% (3/36). The three most common CGG repeats were 29, 30, and 36, respectively. Analysis of AGG interruption pattern in 242 samples identified 908 AGG interruptions, involving 67 different patterns. The most frequent AGG interruption pattern was (CGG)9AGG(CGG)9AGG(CGG)9. Furthermore, long-read sequencing was successfully applied for prenatal diagnosis in two pregnant women, and dynamic mutation of CGG repeat was detected within one family. CONCLUSION: Long-read sequencing-based assay cannot only accurately detect CGG repeat and AGG interruption, but also simultaneously identify other abnormalities of the FMR1 gene. Long-read sequencing offers a broader detection scope and better characterization of FXS-related genetic features.

Humans↗
PubMedDate not supplied

Pheochromocytoma. Cytologic findings on intraoperative scrape smears in five cases.

BACKGROUND: There have been few studies describing the cytology of adrenal pheochromocytoma (PC). Although fine needle aspiration (FNA) for a preoperative diagnosis of PC is generally considered a contraindication, this tumor can be an unsuspected finding in adrenal FNA performed for other reasons. STUDY DESIGN: Scrape cytology smears prepared in five cases of PC were examined for different cytomorphologic features. The results were correlated with the corresponding permanent histologic sections. RESULTS: Previously described features, like cellular smears showing cells with abundant, poorly defined fragile cytoplasm, bare nuclei, anisonucleosis, "salt and pepper" chromatin, variable nucleoli and few ganglion cell-like cells, were noted. In addition, several previously unreported cytologic features were observed: (1) loosely cohesive PC cells along a ramifying, delicate central core; (2) intracytoplasmic microvesicular (not hyaline/homogeneous) globules; and (3) different arrangements of capillary-stroma and PC cells (Zellballen pattern; empty capillary rings; stroma with adherent, intact PC cells or fragments of disrupted PC cell cytoplasm). CONCLUSION: The cytologic appearance of PC may resemble that of other neuroendocrine tumors; however, it can be diagnostic when combined with proper clinical data and ancillary tests.

Adrenal Gland Neoplasms↗
PubMed2001

Structural fragility of blood vessels and peritoneum in calponin h1-deficient mice, resulting in an increase in hematogenous metastasis and peritoneal dissemination of malignant tumor cells.

We have observed weak expression of calponin h1, which stabilizes the actin filament system, in blood vessels within human malignant tumors. This observation suggested that because of a deficiency in stabilization by calponin h1, the structure of blood vessels in malignant tumors is fragile compared with blood vessels in normal tissues. We therefore generated calponin h1-deficient (CN(-/-)) mice to examine the effect of calponin h1 on the integrity of the barrier system in blood vessels against cancer metastasis. The CN(-/-) mice exhibited morphological fragility of the tissues, including the uterus and blood vessels. In particular, we frequently observed bleeding into the surrounding tissue from blood vessels of the ocular fundus in CN(-/-) mice. In addition, mesothelial cells, which usually express calponin h1 in normal (CN(+/+)) mice, were retracted in the CN(-/-) mice. When fluorescein was injected i.v. into mice, the CN(-/-) mice exhibited a greater and more rapid leakage of fluorescein from the blood vessels of the ocular fundus compared with the CN(+/+) mice. In the CN(-/-) mice receiving i.v. inoculations of B16 melanoma cells, significantly more metastatic nodules were formed in the lung than in the CN(+/+) mice. When B16 melanoma cells were injected i.p., the severity of peritonitis carcinomatosa was greater in CN(-/-) than in CN(+/+) mice. These results indicate that calponin h1 plays an important role in the regulation of the integrity of the blood vessels and peritoneum, which in turn is an important factor influencing the frequency of cancer metastasis. The CN(-/-) mice, which exhibit fragile blood vessels and peritoneum, could serve as sensitive and useful host models to investigate cancer metastasis.

Actins↗
PubMed1994

Postnatal retinal vascularization in former preterm infants with retinopathy of prematurity.

PURPOSE: To study the postnatally vascularized retina in former preterm infants in whom retinopathy of prematurity (ROP) stages 2 to 4a developed and spontaneously regressed. METHODS: Matched fundus photographs and fluorescein angiograms of the temporal peripheral retinas of 133 eyes (72 patients) were obtained after 2 years of age (mean, 7.7 years; range, 2-16.2 years) and were quantified by two masked observers with respect to the following parameters: (1) macular ectopia (in disc diameters); (2) vessel traction (in 30 degrees sectors); (3) radial length of postnatally vascularized retina (in disc diameters); and (4) capillary scaffolding of postnatally vascularized retina (as a density). These cicatricial outcomes were then compared with their active worst ROP stage. RESULTS: Of the 133 retinal montages, the following active worst ROP stages had been documented: 30 with stage 2, 42 with stage 3 mild, 32 with stage 3 moderate, 20 with stage 3 severe, and 9 with stage 4a. As active worst ROP stage increased, macular ectopia and vessel traction increased, and radial length and capillary scaffolding of postnatal retinal vascularization decreased. Retinal holes were documented frequently in eyes with high myopia. CONCLUSIONS: The peripheral retina in former preterm infants warrants close scrutiny for possible late rhegmatogenous retinal detachments. Prolonged retinal traction (by remnant shunt and extraretinal fibrovascular proliferation) between stable, posterior, prenatally vascularized retina, and unstable, postnatally vascularized retina may lead to the development of retinal holes characteristically located in the fragile, anterior, undifferentiated, nonvascularized retina.

Adolescent↗
PubMed1995

Study of the intraosseous vessels of the femoral head in patients with fractures of the femoral neck or osteoarthritis of the hip.

Bearing in mind earlier studies which established a link between arteriosclerosis and mineral loss, or fragility of the bones, and also our recent study showing that patients with arterial disorders of the lower limbs also suffered from osteoporosis, we carried out a histological study of the number and appearance of the intraosseous vessels and trabecular bone volume in the femoral heads of patients undergoing surgery for either fracture of the femoral neck or osteoarthritis of the hip. The number of thick-walled vessels, arterioles or arterial capillaries was significantly diminished in the femoral heads of patients with fractures of the femoral neck (p = 0.007). In addition, in the latter patients, arteriosclerotic vascular lesions (rupture of the internal elastic lamina, medial thickening and fibrosis) were more frequent than in patients with osteoarthritis of hip. The possibility that, through chronic ischemia, arteriosclerosis may lead to disturbance of bone remodelling and loss of the mechanical properties of bone has not been contradicted by these findings.

Aged↗
PubMed1981

Red cell flexibility and oxygen affinity in patients with angina pectoris and normal coronary arteries.

Plasma viscosity, haemoglobin-oxygen affinity and red cell flexibility were determined in 16 patients who had angina pectoris but no objective evidence for spasms or obstructions in the coronary arteries. Reference data were obtained from healthy controls and from a group of patients with angina pectoris and multivessel coronary artery disease. In the non-coronary angina group mean plasma viscosity (+/- SD) was in the normal range (1.3 leads to 0.06 centipoise). The same was true for haemoglobin-oxygen affinity (P50: 25.9 +/- 1.7 mmHg). Both red cell rigidity (71 +/- 20 mmHg) and red cell fragility (142 +/- 95 mg/l) were significantly higher (p les than 0.0025) than in the control group (54 +/- 10 mmHg and 63 +/- 29 mg/l, respectively). The group with coronary artery obstructions did not differ from the controls. Thus rigid red cells appear to be related to the syndrome of angina pectoris despite normal coronary arteries. In the light of previous findings of local perfusion abnormalities in these patients, inflexible erythrocytes could be an additional factor compromising myocardial capillary flow.

Adolescent↗
PubMed1997

Age-dependent susceptibility in mumps-associated hydrocephalus: neuropathologic features and brain barriers.

Central nervous system susceptibility to viral infection is often age dependent for unclear reasons. In this study, we examined the age-dependent susceptibility of the brain in mumps virus-induced hydrocephalus in hamsters, and evaluated the relationship between neuropathologic features and brain barriers using glial fibrillary acidic protein and zonula occludentes 1 (ZO-1) immunohistochemistry. In a group intracerebrally inoculated with mumps virus at 2 days of age, pathologic findings such as periventricular edema, ependymal cell loss, and ventricular dilation were more prominent and the distribution of mumps virus antigen was wider than in a group inoculated at 30 days of age. ZO-1-immunoreactive tight junctions in the hydrocephalic brains of the 2-day group were severely damaged in the choroid plexus and ependyma, and in white matter capillaries as early as 3 days after inoculation. These changes were not apparent in the hydrocephalic brains of the 30-day group. Prominent cortical dissemination of virus in the 2-day group was related to underdeveloped perivascular glial foot processes in brain parenchyma. Periventricular edema in the 2-day group was linked to ependymal and blood-brain barrier tight-junction permeability. Our results suggest that tight junctions in the early postnatal period are more immature and fragile than in the adult. We concluded that brain susceptibility in mumps virus-induced hydrocephalus is intimately related to the maturity of brain barriers.

Aging↗
PubMed1992

[Clinicopathological study on progressive hereditary nephritis: observations of ultrastructural lesions in the glomerular basement membrane].

Four boys and six girls with progressive hereditary nephritis were studied clinicopathologically. Renal biopsy was performed 16 times in ten cases. Mean age at renal biopsy was 7.3 years old (range 2 to 14 years old). The obtained results were as follows: (1) Montages of electron micrographs were prepared to complete one whole glomerulus. The length of the glomerular basement membrane (GBM) with the characteristic splitting of the lamina densa (Reticulation) was measured and expressed as a percentage of the total length of the GBM. The range of the percentage of the GBM with Reticulation was from 2 to 43% (13.4 +/- 10.0%, mean +/- SD, n = 16). In 4 cases of the 5 cases performed serial renal biopsy, the percentage of the GBM with Reticulation at the 2nd biopsy increased compared with the 1st one. (2) Protein excretion in the urine, serum albumin, alpha 2-globulin, fibrinogen and total cholesterol showed the correlation with the percentage of the GBM with Reticulation. (3) Incomplete ruptures (deep invasion of the epithelial cells into the thickened GBM with Reticulation) were observed. Those suggested that the GBM became fragile associated with the expansion of Reticulation and finally ruptured. Gaps of the GBM were observed 0 to 3 per in one glomerulus (0 to 1.87 per 1mm GBM) and the serial biopsies showed an increase in the number of the gaps as time passed. (4) This study showed the increase in factors activating the blood coagulation such as total cholesterol and fibrinogen, with the expansion of the GBM with Reticulation. And in a nephrotic case, fibrin strands were observed in the glomerular capillary loops and in the GBM. These findings suggest that the activation of the blood coagulation plays a role for the damage of the glomeruli in progressive hereditary nephritis.

Adolescent↗
PubMedDate not supplied

[Hormones and venous system].

Sex hormones have an effect on venous "content" and "container" according to their chemical nature, their dosage and their mode of administration: 17 beta-estradiol (endogenous): protective effect; synthetic estrogens, at normal or low doses: thrombogenic; oral natural estrogens: thrombogenic; extra-digestive natural estrogens: non thrombogenic; non steroid progestagens (androgenic): thrombogenic; non androgenic progestagens: non thrombogenic. Clinically, the venous disease si characterized by sudden episodes occurring at key-periods of the hormonal life: puberty, pregnancy, menopause, oral contraceptives intake, substitute treatments of menopause, premenstrual syndrome. Evaluation of these different situations shows that an early treatment is possible and needed, which, although not providing a new venous wall for these constitutionally fragile patients, may act effectively at two levels: 1) correction of the haemodynamic disorder (venous reflux in the saphenous arches and the perforators; 2) resorption of tissue infiltration. As primary prevention, in a patient with hormonal disorders or who must be treated with estrogens or progestagens, the objective of our treatment is to protect the venous wall and encourage the return circulation. One must: 1) reinforce the vaso-constrictive effect and the parietal tone, 2) limit collagen and elastin alteration, 3) reinforce capillary permeability and decrease the interstitial edema, 4) normalize the haemorheological constants, 5) restore the balance hemostasis-fibrinolysis. The opinion of a phlebologist seems essential before prescribing a hormonal treatment and monitoring the effects of the treatment. Cooperation between gynaecologists and phlebologists is particularly essential in the interpretation of the clinical disorders as well as discussing the venous risk, the dosage and the administration route of sex hormones.

Catecholamines↗
PubMed1990

Cause of thrombosis in human atherosclerotic arteries.

Although it has been known for a long time that thrombosis nearly always develops in atherosclerotic arteries--and almost never in normal vessels--the mechanism through which atherosclerosis promotes thrombosis was unknown until this problem was explored through histologic examination of complete serial section sets of thrombosed atherosclerotic arteries. These studies, repeatedly confirmed, revealed that the thrombi are triggered by microscopic cracks in the collagen cap of advanced plaques. Blood most often seeps from the arterial lumen through the cracks into the underlying lipid gruel before the breaks or cracks are plugged by the thrombi (which function as hemostatic seals of the breaks). These results are parallel to results of experimental studies in which the synergism of endotheliotoxic and pressor agents produced thrombi over cap breaks and sub-break hemorrhages only in arteries with advanced collagen-rich plaques, not in arteries with early atherosclerosis or in normal vessels. This finding indicates that advanced atherosclerosis makes the arterial wall much more fragile and that, once broken, this wall exposes the blood to powerful thrombogenic materials that do not exist in normal arterial tissue. At present, human and experimental evidence suggest that the thrombogenic fissures of advanced plaque caps can be promoted by several factors, such as a surge in intraarterial pressure or insults that damage the caps structurally and increase their vulnerability to any type of stress such as certain metabolic, exogenous chemical and immune insults, spontaneous molecular changes of collagen with time and hemorrhages of capillaries that invade advanced plaques from the adventitia or the arterial lumen.

Capillaries↗