Disruption of the Golgi apparatus by brefeldin A blocks cell polarization and inhibits directed cell migration.

The role of the Golgi apparatus in the motile activity of fibroblasts was examined with brefeldin A (BFA), which disrupts the Golgi apparatus in a variety of cells. Upon incubation with BFA, Swiss mouse 3T3 fibroblasts lost their typical polarized morphology, in which the leading edge is characterized by intensive lamellipodia formation. BFA affected cell asymmetry as demonstrated by a decrease in the morphometric indices, dispersion, and elongation. After BFA treatment, cells showed little protrusional activity and did not form a dense actin network at the leading edge, and consequently the rate of cell migration into an experimental wound was significantly reduced. In addition, BFA prevented an increase in pseudopodial activity and prevented the formation of long processes induced by phorbol 12-myristate 13-acetate. The effects of BFA on cell shape and protrusional activity were quantitatively similar to those observed with the microtubule-disrupting agent nocodazole, although BFA had no effect on microtubule integrity. These results suggest that the integrity of both the Golgi apparatus and microtubules is necessary for the generation and maintenance of fibroblast asymmetry, which is a prerequisite for directed cell migration.     

Light microscopic morphometric analysis of rat ileal mucosa

A light microscopic morphometric analysis of IgA-containing immunocytes within samples of ileal mucosa was performed. The following groups of rats were studied: (1) animals raised in a gnotobiotic environment (microbial reduction); (2) animals with iatrogenic self-filling intestinal blind loops (microbial proliferation); and (3) control animals (sham operation). The unlabeled antibody enzyme immunohistochemical localization technique was employed for the identification of intracellular IgA. Component quantitation involved use of a micrometer component quantitator. Numerical density of the immunocyte population was determined by component quantitation of individual and total immunocyte volumes and by application of the Floderus equation. The methodology employed provided a precise quantitative analysis of all mucosal components of normal and manipulated rat ileum. A statistically significant reduction in the volume percentage of IgA-containing immunocytes in association with both microbial reduction and microbial proliferation was observed. The volume percentage reduction of the IgA-containing immunocyte population associated with gnotobiosis may reflect decreased microbial antigenic stimulation, whereas that associated with microbial proliferation may reflect the presence of an increased population of immunocytes producing non-IgA immunoglobulins.     

老年糖尿病人甲襞微循环改变与眼底血流动力学变化

目的　观察老年 2型糖尿病 (2DM)病人甲襞微循环改变及眼底动脉血流动力学变化并探讨其临床意义。　 方法 　对 40例老年 2DM病人 (DM组 )及 3 0例健康老年人 (对照组 )行甲襞微循环观测并用彩色多普勒超声显像 (CDFI)检测其双眼底动脉血流动力学变化。　 结果 　DM组的管袢变细 ,畸形增多 ,其数目、形态、血液流态及袢周状态的分值及总积分明显高于对照组 ,而血液流速显著低于对照组 (P <0 0 1) ;DM组眼底血流的脉搏指数 (PI)及阻力指数 (RI)高于对照组 (P <0 0 1)。　 结论　老年DM病人微循环明显障碍 ,可作为诊治其并发症及判断疗效的筛选指标 ;老年DM病人眼底血流PI及RI与眼底病变有极高相关性 ,可作为早期发现眼底微血管病的一种可靠方法。     

Component quantitation of rat ileum

Precise and accurate light microscopic morphometric analyses of biological tissue can be achieved utilizing component quantitative techniques. Component quantitation refers to measurements of the relative volumes of components in tissue sections. Such an assessment is predicated upon the mathematically verifiable assumption that direct quantitative relationships exist between an aggregate of profiles of a component contained per unit area in multiple sections and an aggregate of profiles contained per unit volume. A linear scanning device (micrometer component quantitator) was initially employed for quantitative analyses of pancreas. This quantitative technique has subsequently been applied to normal rat ileum conventionally processed for light microscopy, and the requisite sampling parameters have been defined. An identical technique was then applied to physiologically manipulated rat ileum—a gnotobiotic group, a group with ileal self-filling blind loops, and a group with ileal Thiry-Vella loops. The results observed support the following conclusions. (1) The volume percentage of the various components of the rat ileal wall of control animals was defined utilizing the micrometer component quantitator. (2) Hypertrophy of the ileal muscularis externa within the ileal self-filling blind loops was observed, probably secondary to mechanical obstruction. (3) Atrophy of the ileal epithelium within the gnotobiotic group and within the Thiry-Vella loops was observed, possibly secondary to an altered endogenous microbial flora. (4) Recognition of quantitative variations among the histological components of the intestinal wall in association with physiological manipulations or pathologic states was (is) feasible by utilization of this component quantitative technique.A portion of this paper was presented from platform at the 97th Annual Session of the American Association of Anatomists, Seattle, Washington, April 8–12, 1984.     

Double orifice mitral valve; a coincidental finding.

A double orifice mitral valve (DOMV) represents a rare congenital malformation characterised by two valve orifices with two separate subvalvular apparatus. This case demonstrates the necessity of careful imaging of the mitral valve apparatus, not only in patients with atrioventricular septal defects, but also in patients with congenital left obstructive heart disease.     

A method to detect microfilariae of Dipetalonema witei in the peripheral blood of mastomys natalensis (author's transl)]

A new cheap and time saving procedure is described for the detection of microfilariae of Dipetalonema witei in the peripheral blood of Mastomys natalensis. A special apparatus delivering an ether-air mixture is used to anesthetize the experimental animals for a short period in a very careful way and without side effects. The ether wapour induces a rapid shift of microfilariae within one minute from the lung to the peripheral blood.     

Interleukin-13 in systemic sclerosis: relationship to nailfold capillaroscopy abnormalities

 The aim of this study was to investigate whether interleukin-13 (IL-13) serum levels correlate to different nailfold capillaroscopy (NC) findings in patients with systemic sclerosis (SSc). IL-13 serum levels were measured using an ELISA method. The following NC abnormalities were considered: the presence of giant loops, haemorrhages, loss of capillaries, disorganisation of the vascular array, ramified/bushy capillaries and sludging of blood. A semiquantitative rating scale was adopted to score these changes, as well as a rating system for avascular areas and three morphological NC patterns (`early', `active' and `late'). Mean capillary density was determined by counting the total number of capillaries in a 1 mm length, and the arterial and venous diameters of the capillary as well as the total loop diameter were measured. In SSc patients IL-13 serum levels were significantly higher than in controls (P < 00.1), whereas in patients with (n=8) and without (n=24) abnormal IL-13 serum levels (>17 pg/ml) the comparison of the NC features showed significantly relevant differences concerning a more frequent `active' NC pattern (P < 0.02), the presence of haemorrhages (P < 0.0037) and sludging of blood (P < 0.038), as well as larger total loop (P < 0.036) and arterial (P < 0.03) diameters, in those patients with elevated IL-13 serum levels. The study confirmed that IL-13 serum levels are higher in the sera of patients with SSc, and shows for the first time the significant correlations between this serological finding and some of the main relevant SSc capillaroscopic features, leading us to believe that this cytokine not only seems to sustain the immunological and fibrotic process of SSc, but might have a role in determining the more severe microvascular lesions in this disease. Received: 15 July 2002 / Accepted: 16 October 2002     

Modifications of pancreatic islet tissue after internal biliary fistula (experimental research)

Summary The morphometric study of the endocrine pancreas carried out in normal dogs and in dogs with a total internal biliary fistula (cholecystojejunostomy after closure of the common bile-duct) has shown that, following this operation, obvious and statistically significant changes take place which indicate a considerable increase of the insular mass. This anatomic modification, probably induced by the suppression of bile flow into the duodenum, is the result both of hypertrophy and hyperplasia of the endocrine pancreatic apparatus and seems to go hand in hand with an increase in insulinopoietic activity.     

Functional morphology of the adrenal cortex and incretory kidney structures in the salt-losing form of adrenogenital syndrome in children

The adrenals and kidney incretory structures (the juxtaglomerular apparatuses and renomedullary interstitial cells) were studied in 12 cases of a desalinization form of the adrenogenital syndrome in children who died at the age of 1-6 mos of acute water-electrolytic disturbances in order to assess function of the structures involved in the regulation of the water-salt equilibrium. The adrenals and kidneys from 7 children of the same age who died of mechanical asphyxia, were taken as controls. The depth of adrenocortical zones, volumes of nuclei and nucleoli in different cortical zones were determined; the juxtaglomerular index, cell count (including vacuolized cells), the area of the juxtaglomerular apparatus and mesangium were determined in the juxtaglomerular apparatus. The amount of cell lipid granules (the interstitial-cellular index), the mean granule volume and the total granule volume were counted in renomedullary interstitial cells. The results were statistically processed, and a paired correlation analysis of all the studied parameters was performed. In the desalinization form of the adrenogenital syndrome morphometric investigation of the glomerular and fascicular zones revealed compensatory processes, not of hyperplastic nature but mainly of hypertrophic nature; morphological and morphometric characteristics of the kidney incretory structures indicated tension of the renin-angiotensin system and probably a decrease in prostaglandin synthesis in salt and water loss. In the control group parameters of structures related to the regulation of the water-salt equilibrium, especially in the adrenal cortex, showed good correlation. In the desalinization form of the adrenogenital syndrome some other relationships between these structures developed.     

Lysosomes and pancreatic islet function: adaptation of beta-cell lysosomes to various metabolic demands

The effect of various functional demands on the lysosomes of pancreatic islet beta cells was studied in vivo. To expose pancreatic islets to different metabolic situations, normal syngeneic mouse islets were transplanted to either lean mice, alloxan-diabetic mice, or obese hyperglycemic mice. Two weeks after transplantation, primary and secondary beta-cell lysosomes of the islet grafts were analyzed by morphometry. The beta-cell lysosomes and secretory granules of the endogenous islets of lean and obese hyperglycemic mice were also measured. The beta cells of the islets transplanted to lean normoglycemic mice showed only a moderately developed synthetic apparatus and a great number of secretory granules. They had mainly secondary lysosomes, frequently containing secretory granule material, indicating a high crinophagic activity. The islet beta cells exposed to the high serum glucose concentration of alloxan-diabetic and obese hyperglycemic mice had an extensive synthetic apparatus, but a very small content of secretory granules. In these beta cells, there was a predominance of small primary lysosomes, indicating a low crinophagic activity. It is concluded that crinophagy may provide a mechanism for the pancreatic beta cell to moderate its content of insulin. When its secretory granule stores are diminished due to increased demands on insulin secretion, the beta cell seems able to drastically decrease the crinophagic activity. The detailed morphometric analysis of the endogenous islets of the lean and obese hyperglycemic mice showed that the beta cells of the obese hyperglycemic mice had a smaller number and size of the secretory granules.(ABSTRACT TRUNCATED AT 250 WORDS)     

Juxtaglomerular apparatus T-cell infiltration affects glomerular structure in Type 1 diabetic patients

Aims/hypothesis Type 1 diabetes is an autoimmune disorder associated with T-cell mediated injury to multiple endocrine tissues. T-cell infiltration of the juxtaglomerular apparatus could be associated with changes in local renin angiotensin system activity and, thus, with changes in the renal microenvironment. We examined the frequency of juxtaglomerular apparatus T-cell infiltration early in Type 1 diabetes and tested whether this is associated with renal structure and function.Methods We classified 89 Type 1 diabetic patients by immunohistochemical analysis as either juxtaglomerular apparatus T-cell positive (n=37) or T-cell negative (n=38). Borderline cases (n=14) were not considered further.Results T-cell positive patients had a shorter duration of diabetes (6.7±2.5 years) than T-cell negative patients (9.2±5.0 years, p=0.011) and lower albumin excretion rate, but they had a similar glomerular filtration rate and blood pressure. Renal biopsy morphometric analysis showed similar glomerular basement membrane width and mesangial fractional volume in these two groups. However, glomerular capillary surface density (p=0.0012) and filtration surface per glomerulus (p=0.0155) were greater in the T-cell positive patients.Conclusion/Interpretation Increased filtration surface per glomerulus could be associated with glomerular filtration rate preservation in diabetes. Thus, juxtaglomerular apparatus immunologic injury in Type 1 diabetes patients could delay the clinical consequences of diabetic nephropathy.Abbreviations ECM Extracellular matrix - RAS renin-angiotensin-system - JGA juxtaglomerular apparatus - NHDN natural history of diabetic nephropathy - IP immunoperoxidase - GV mean glomerular volume - TGF-B transforming growth factor-B - Vv(Mes/glom) mesangial fractional volume per glomerulus - GBM glomerular basement membrane - Sv(PGBM/glom) surface density of the peripheral glomerular capillary perglomerulus - TFS total filtration surface per glomerulus - Vv(Int/cortex) fractional volume of renal cortical interstitium See the Acknowledgements for the list of investigators of the International Diabetic Nephropathy Study Group (IDNSG)     

Repair of single-stranded loops in heteroduplex DNA transfected into mammalian cells.

Repair of heteroduplex DNA, generated between two interacting DNA molecules during homologous recombination, has been implicated as a contributing factor in the process of gene conversion. To assess patterns of heteroduplex repair in mammalian cells, we constructed 13 different heteroduplexes from simian virus 40 wild-type and deletion mutant DNAs. Each heteroduplex contained one or multiple single-stranded loops in the intron of the gene for large tumor antigen, which is not essential during lytic infection. After transfection into cultured monkey cells, cellular repair was evaluated by restriction analysis of the amplified viral progeny from 1123 individual plaques, each representing the clonal expansion of a single repair event. Single-stranded loops were corrected prior to replication with an overall efficiency of 90%. At the position of a loop, one of the two heteroduplex strands served as a template for accurate repair 98% of the time. Repair of single-stranded loops was biased nearly 2 to 1 in favor of the strand without the loop. The efficiency, accuracy, and strand bias of repair were unaffected by loop size within the tested range, which was 25-247 nucleotides. The excision tract associated with repair of single-stranded loops rarely exceeds 200-400 nucleotides in length.     

Cyclophosphamide treatment improves microvessel damage in systemic sclerosis

The objective of this study is to evaluate nailfold videocapillaroscopy changes in scleroderma patients treated regularly on cyclic basis with iloprost and to find associations with clinical, serologic, and pharmacological variables. Forty-nine patients affected by systemic sclerosis (44 women and five men, mean age 52.4 years, mean disease duration 8.0 years, 31 patients with limited cutaneous subset and 18 with diffuse cutaneous form of the disease) underwent two nailfold videocapillaroscopies at a distance of 3 years from each other; the examinations were performed by an operator blinded to clinical features and to drug treatment. Six patients showed an amelioration of nailfold videocapillaroscopic abnormalities who changed from active to early pattern; five of these cases (83.3%) had been given cyclophosphamide therapy and the remaining case methotrexate plus azathioprine. Cyclophosphamide administration was significantly associated with amelioration of nailfold videocapillaroscopic pattern (p < 0.001). None of the patients who received cyclophosphamide demonstrated worsening of the microvascular lesions; the progression of nailfold videocapillaroscopic pattern was inversely correlated to cyclophosphamide treatment (p = 0.02). In our study, cyclophosphamide treatment demonstrated to be effective for scleroderma microvascular damage as directly observed by nailfold videocapillaroscopy.     

Circulating bile salt-dependent lipase originates from the pancreas via intestinal transcytosis

BACKGROUND & AIMS: Bile salt-dependent lipase (BSDL) has been detected in human blood, where it is assumed to play a substantial role in atherosclerosis. The origin of this circulating enzyme is unknown. The aim of this study was to show that blood BSDL originates from pancreatic exocrine secretions via a transcytotic motion across the intestinal epithelium. METHODS: Fluorescein isothiocyanate- or [(125)I]-labeled human pancreatic BSDL was instilled into the lumen of intestinal loops of the rat, and combined biochemical and immunocytochemical investigations were performed in intestinal cells and in the blood of these animals. RESULTS: In vivo pancreatic BSDL is internalized by duodenal enterocytes. The pancreatic enzyme was associated with microvilli and present in endocytic vesicles and Golgi apparatus as well as along the basolateral membrane of enterocytes. It was also detected in intestinal interstitial spaces. Radiolabeled pancreatic BSDL internalized by the duodenal epithelium is the one further detected in circulation. The radiolabeled protein was immunoprecipitated from plasma and had a 100-kilodalton molecular mass compatible with native pancreatic BSDL. In blood, BSDL was mainly associated with low-density lipoproteins. CONCLUSIONS: These in vivo data show that BSDL, normally present in blood, originates from exocrine pancreatic secretion and support the pathophysiologic relevance of BSDL transcytosis through the intestinal mucosa cell lining. Based on this, the implication of circulating BSDL in atherosclerosis merits careful attention.     

Erythrocyte aggregation in vivo and diabetic microangiopathy. Physiopathologic determinants and clinical correlations

Conjunctival angioscopy is a non invasive way for visualization and quantitative evaluation of the microcirculation. Grading red blood cell aggregation is possible in vivo using Ditzel's four grades score. The two-dimensional organization of conjunctival microvessels allows morphometric quantification of microvascular density. The aim of our study was to evaluate the determinant factors of erythrocyte aggregation and the place of conjunctival angioscopy in the early detection of diabetic microangiopathy. Conjunctival parameters were red blood cell aggregation (Ditzel's score) and morphometric evaluation of capillary, venular and arteriolar density. Criteria of diabetic microangiopathy were microalbuminuria and retinal fluorescein angiogram. Both conjunctival angioscopy and retinal angiogram were scored independently. Results obtained in 30 type 1 diabetic patients, with multifactorial statistical analysis, show that red blood cell aggregation in vivo is an essential discriminant factor for diabetic retinopathy and nephropathy. This rheological phenomenon depends more on duration of diabetes (analysis of variance p = .0005, r = .65) and metabolic control than on albuminemia or fibrinogenemia such as in non-diabetic patients. Manual morphometric data confirmed vascular rarefaction associated with excessive red blood cell aggregation (beta = .50). These results suggest that grading red blood cell aggregation in vivo is an interesting tool for physiopathological and clinical studies of diabetic microangiopathy.     

Double-Sided Metasurface Array for a Dual-Band and Polarization-Independent Microwave-Energy-Harvesting System

In this article, we present a simple and novel design of a double-sided metasurface for a dual-band and polarization-independent microwave-energy-harvesting system. The proposed metasurface is constructed from the dual-sided design of 8 × 8 unit cells. Different from the regular dual-band unit cells that contain two loops or multiple shapes of resonators printed in the same layer, the proposed metasurface is based on designing double loops, each combined with two arms of a dipole printed on the top and bottom sides of a single substrate. Thus, the bottom layer is utilized to generate the second frequency band of interest. Three main numerical simulations were conducted to investigate the performance of a single unit cell, a 2 × 2 supercell, and an array of an 8 × 8 metasurface structure. The numerical simulation demonstrated that 98% and 95% of the incident energy is collected at two bands of 1.8 and 6.5 GHz for the proposed harvester.     

Morphometric analysis of the parathyroid glands and thyroid gland C-cells during prolonged administration of triiodothyronine and mercazolyl

The aim of the study was to perform a systemic morphometric analysis of the parathyroid glands and parafollicular thyrocytes under the conditions of the suppression of function of the thyroid follicular apparatus. Water suspensions of triiodothyronine and methniasole were administered through a probe for 24 days to adult albino male rats (6 in each group) at daily doses of 50 micrograms/kg and 6 mg/kg respectively; 6 animals were controls. The level of total calcium in the blood serum of animals in the study groups did not differ from control values. The administration of the drugs induced the development in the parathyroid glands of morphological signs of their increased function (hypertrophy of the parathyrocytes, their nuclei and cytoplasm in both series of experiments, an increase in the mean count of nucleoli in the nucleus in the administration of triiodothyronine). Signs of elevated secretory activity of the population of parafollicular thyrocytes (a decrease in the relative frequencies of C-cells with the cytoplasm filled in with secretory granules; an increase in the proportion of cells with cytoplasmic zones at the vascular poles free of granules) were detected in the thyroid.     

Application of Morphometric Analysis to Identify Alewife Stock Structure in the Gulf of Maine

Abstract

Alewife Alosa pseudoharengus is an anadromous clupeid fish of long-standing ecological and socioeconomic importance along the Atlantic coast of North America. Since the 1970s, Alewife populations have been declining throughout the species’ range. A number of hypotheses have been proposed to explain the decline, but a lack of basic information on population demographics inhibits hypothesis testing. In this study, we evaluated the use of morphometric analysis to discriminate among spawning stocks of Alewives collected from 24 sites in Maine and one site in Massachusetts. We first identified 10 morphometric measurements that were not influenced by the freezing–thawing process, and then used principal component and discriminant function analyses to develop stock-structure classification models from these 10 measurements. Classification models were able to discriminate Alewives to be from Maine or the single Massachusetts site 100% of the time. In addition, classification models correctly classified pooled sampling sites from the extreme western and eastern parts of Maine with 64% accuracy. Morphometric analysis may therefore provide an easily accessible, comparatively fast, and inexpensive method to discriminate marine-captured Alewives spawned in areas separated by major biogeographic regions, large geographic distances (100s of kilometers), or both, and thus help inform questions about stock composition at these spatial scales for assessment surveys and bycatch events.

Received March 30, 2012; accepted October 14, 2012.