Capillary Loops Research Articles

Dermatoscopic assessment of nailfold capillary abnormalities in Behçet's disease and correlation of the findings with disease activity and severity

Background: A small number of studies used nailfold capillaroscopy (NC) in the evaluation of nailfold capillary changes in Behçet's disease (BD). The purpose of this study was to investigate the characteristics and frequency of nailfold capillary changes in BD by dermatoscopy and videodermatoscopy and to develop a scoring system for those capillary changes to predict the activity and severity of the disease. Methods: We performed NC on 40 BD patients and 20 healthy controls with dermatoscopy and videodermatoscopy. Capillary morphology, distribution, and density were analyzed qualitatively and quantitatively. We also assessed the activity and severity of the disease with BD Current Activity Form and BD Severity Score Classification and evaluated the relation of these scores with morphology scores and capillary density. Results: Capillary morphologic alterations were encountered significantly more in BD group (P < 0.05). Loss of continuity in capillary loops and irregularity of capillary distribution were significantly more frequent in BD group compared to healthy controls (P = 0.003 and P < 0.001). Morphology score was significantly higher in BD patients compared to control group (P < 0.001); however, we could not detect a significant relation between capillary morphology and density and the activity and severity of BD. Conclusion: Although we could not demonstrate a significant relation between capillary changes and the activity and severity of BD, we consider that NC performed with dermatoscopy can reflect the presence and extent of microvascular involvement and thereby might have diagnostic and prognostic value in BD.

Dermatomyositis: A dermatological perspective

Dermatomyositis (DM) is an autoimmune idiopathic inflammatory myopathy which has distinctive skin lesions, specific autoantibodies, and most importantly, symmetrical proximal muscle weakness and muscle inflammation of subacute onset. Some cases of DM present with only characteristic skin lesions without muscle disease and referred to as clinically amyopathic DM. Characteristic skin lesions include violaceous discoloration around the eyelids (heliotrope rash), erythematous rash over the face, erythematous scaly papules over the interphalangeal and metacarpophalangeal joints (Gottron's papules), erythema over the same joints (Gottron's sign), photosensitive pruritic erythema over the anterior chest and neck (V-sign), erythema extending to the shoulders and back (Shawl sign), distorted cuticles with dilated capillary loops over the proximal nail folds, and hyperkeratotic fissured skin over the palms and lateral aspect of the fingers (Mechanic's hands). Causes for mortality and morbidity in DM include malignancy, respiratory disease, and cardiac disease. After ruling out internal associations, Mangement involves photoprotection and medications including systemic corticosteroids, antimalarials, mycofenolate mofetil, biologicals and IV immunoglobulins, depending upon patient parameters.

THE VALUE OF NARROW BAND IMAGING ENDOSCOPY IN DIAGNOSIS OF HYPOPHARYNGEAL AND LARYNGEAL CANCER AND FOLLOWING - UP POST TREATMENT

Background: To evaluate the value of narrow band imaging (NBI) endosocopy in diagnosis of hypopharyngeal and laryngeal cancer and following – up post treatment. Material and methods: A total of 75 patients included 36 patients with hypopharyngeal cancer and 39 patients with laryngeal cancer who had diagnosed at Department of Otoloryngology – Hue Central Hospital from 5/2017 to 5/2018. A prospective cohort study was conducted. Results: The age group 51 - 60 years occurred most often, 33.3%, the mean age was 62.1 ± 13.4. The UICC stage III was 65.3%. Tumor was in ulcerlarative and infiltrate form (89.4%), edema and inflammation of magrin tumor (41.3%), invasive (58.7%). Intrapapillary capillary loops – IPCL - type V was predominant, type V-n was 46.7%. The tumor with IPCL type V-n had strong enhancement (51.3%) and moderate enhancement (44.4%) after contrast medium injection on CT scan. One month after treatment, there were 33.3% of tumor – free, 53.7% of mucosal edema and 13% tumor size-decreasing on NBI image. Conclusion: NBI endoscopy is an useful tool for diagnosing of hypopharyngeal and laryngeal cancer and following – up post treatment. Key words: narrow band imaging endoscopy, hypopharyngeal cancer, laryngeal cancer

Computer-assisted diagnosis of early esophageal squamous cell carcinoma using narrow-band imaging magnifying endoscopy.

We developed a computer-assisted diagnosis model to evaluate the feasibility of automated classification of intrapapillary capillary loops (IPCLs) to improve the detection of esophageal squamous cell carcinoma (ESCC). We recruited patients who underwent magnifying endoscopy with narrow-band imaging for evaluation of a suspicious esophageal condition. Case images were evaluated to establish a gold standard IPCL classification according to the endoscopic diagnosis and histological findings. A double-labeling fully convolutional network (FCN) was developed for image segmentation. Diagnostic performance of the model was compared with that of endoscopists grouped according to years of experience (senior > 15 years; mid level 10 - 15 years; junior 5 - 10 years). Of the 1383 lesions in the study, the mean accuracies of IPCL classification were 92.0 %, 82.0 %, and 73.3 %, for the senior, mid level, and junior groups, respectively. The mean diagnostic accuracy of the model was 89.2 % and 93.0 % at the lesion and pixel levels, respectively. The interobserver agreement between the model and the gold standard was substantial (kappa value, 0.719). The accuracy of the model for inflammatory lesions (92.5 %) was superior to that of the mid level (88.1 %) and junior (86.3 %) groups (P < 0.001). For malignant lesions, the accuracy of the model (B1, 87.6 %; B2, 93.9 %) was significantly higher than that of the mid level (B1, 79.1 %; B2, 90.0 %) and junior (B1, 69.2 %; B2, 79.3 %) groups (P < 0.001). Double-labeling FCN automated IPCL recognition was feasible and could facilitate early detection of ESCC.

Glomerular basement membrane duplication is a predictor of the prognosis of diabetic nephropathy in patients with type 2 diabetes.

Several clinical parameters and pathological findings are known to be predictors of the deterioration of diabetic nephropathy (DN). Glomerular basement membrane duplication (GBM-DP) is a pathological feature representing endothelial injury, which is commonly observed in DN. In the present study, we investigated the association between GBM-DP and the renal prognosis in DN. The study enrolled 80 patients with renal biopsy-proven DN who were managed at Chiba-East Hospital from 2005 to 2012. We confirmed the pathological findings according to the Renal Pathology Society classifications, and we further evaluated the GBM-DP, which was defined as double contours of the GBM that expanded more than 10% of capillary loops in the most affected nonsclerotic glomerulus. We used Cox regression models to estimate hazard ratios (HRs) for end-stage renal disease (ESRD), with adjustment for age, sex, systolic blood pressure, HbA1c, estimated glomerular filtration rate (eGFR), and urinary protein excretion (UP) at baseline. Of the 80 patients, 56 were male (70.0%) and the mean age was 59.1years. The median eGFR and UP were 42ml/min/1.73m2 (IQR 30, 59) and 3.1g/gCr (IQR 1.2, 5.2). Twenty-seven patients progressed to ESRD and one patient died during the median observational period of 2.9years (IQR 1.5, 4.3). The multivariable analyses showed that GBM-DP was significantly associated with ESRD (HR 3.18 [95% confidence interval (CI): 1.02-9.87], p = 0.045). We newly identified GBM-DP as a strong prognostic predictor in DN patients. Further study is needed to clarify the pathogenic mechanism of GBM-DP in DN.

Early identification of esophageal squamous neoplasm by hyperspectral endoscopic imaging

Esophageal squamous neoplasm presents a spectrum of different diatheses. A precise assessment for individualized treatment depends on the accuracy of the initial diagnosis. Detection relies on comprehensive and accurate white-light, iodine staining, and narrow-band imaging endoscopy. These methods have limitations in addition to its invasive nature and the potential risks related to the method. These limitations include difficulties in precise tumor delineation to enable complete resection, inflammation and malignancy differentiation, and stage determination. The resolution of these problems depends on the surgeon’s ability and experience with available technology for visualization and resection. We proposed a method for identifying early esophageal cancerous lesion by endoscopy and hyperspectral endoscopic imaging. Experimental result shows the characteristic spectrum of a normal esophagus, precancerous lesion, canceration, and intraepithelial papillary capillary loop can be identified through principal component score chart. The narrow-band imaging (NBI) image shows remarkable spectral characteristic distribution, and the sensitivity and specificity of the proposed method are higher than those of other methods by ~0.8 and ~0.88, respectively. The proposed method enables the accurate visualization of target organs, it may be useful to capsule endoscope and telemedicine, which requires highly precise images for diagnosis.

The added value of magnifying endoscopy in diagnosing patients with certain gastroesophageal reflux disease

PurposeIn most cases gastroesophageal reflux disease proceeds without macroscopic erosions in the esophagus. We aimed to clarify if abnormalities detectable in magnifying endoscopy may offer additional diagnostic criteria for gastroesophageal reflux disease and to what histopathologic structures do they correspond. Patients/methodsEsophageal mucosa above and below Z-line was evaluated under x115 magnification in 67 gastroesophageal reflux disease patients (11 with erosive reflux disease, 28 with Barrett’s esophagus, 28 with nonerosive reflux disease) and in 12 patients without gastroesophageal reflux disease (negative control group). Features characteristic of gastroesophageal reflux disease were specified by comparing erosive reflux disease and Barrett’s esophagus patients with negative control group. Afterwards the presence of identified features were evaluated in nonerosive reflux disease group. Interobserver agreement in the recognition of the proposed criteria was rated. Biopsies collected from the mucosa above Z-line were evaluated histologically after hematoxylin and eosin staining. ResultsEndoscopic lesions characteristic of gastroesophageal reflux disease were: microerosions, abnormal intrapapillary capillary loops, obscured palisade vessels, white points, big triangular indentations of Z-line and villous mucosa below Z-line. The presence of two or more of the above features indicated gastroesophageal reflux disease with 97% sensitivity and 75% specificity. Substantial interobserver agreement was achieved in evaluation of obscured palisade vessels, abnormal intrapapillary capillary loops and white points. Endoscopic lesions were correlated to histology. Lesions identified with magnifying endoscopy were helpful in discerning between negative control group and nonerosive reflux disease patients. ConclusionsMagnifying endoscopy reveals abnormalities that can be used as additional endoscopic diagnostic criteria of gastroesophageal reflux disease.

A Cross-Sectional Observational Study of Nailfold Capillary Morphology in Uveitis

ABSTRACTPurpose: We performed nailfold capillary microscopy to explore microvasculature abnormalities in uveitis overall and uveitis stratified in various ways.Methods: This was a cross-sectional, case–control, observational study. One hundred and seven uveitis patients and 130 control subjects were included. We used a JH-1004 capillaroscope to perform nailfold capillary video microscopy on the fourth and fifth digits of each subject’s nondominant hand. Videos were evaluated for hemorrhages, dilated capillary loops > 25 µm, and avascular zones > 200 µm. Univariate analyses were used for the assessment of case–control morphological differences and multivariate analyses were performed to assess the relation between nailfold capillaroscopic findings and uveitis subgroups.Results: In univariate analysis, uveitis patients were more likely to have higher tortuosity ratings and reduced capillary density compared to controls (p < 0.001 for both); furthermore, dilated capillary loops, avascular zone and hemorrhages were more frequent in uveitis versus control subjects (p < 0.001 for all). Among cases, every unit increase in capillary density (vessels/mm) was associated with active uveitis (n = 72 cases) versus inactive disease (n = 35 cases; odds ratio (OR) = 1.7; (95% confidence interval (CI), 1.2–2.5) in multivariate analysis. Furthermore, the presence of any nailfold hemorrhage versus the absence of hemorrhage was more likely to be associated with posterior and panuveitis (n = 41 cases combined) compared to anterior and intermediate uveitis (n = 66 cases combined; OR = 5.8; 95% CI, 2.3–14.2). Moreover, we found a positive correlation between peripheral retinal leakage and nailfold capillaries dilation (r = 0.33; p = 0.015) that was not strictly significant based on the number of comparisons made.Conclusions: Our study provides support for non-ocular capillary bed abnormalities in uveitis, with interesting correlations based on disease stage and anatomical classification.

Magnifying Endoscopy with Narrow Band Imaging in Estimating the Invasion Depth of Superficial Esophageal Squamous Cell Carcinomas

Background/Aim: The aim of this study was to compare the accuracy of magnifying endoscopy with narrow band imaging for the diagnosis of depth of invasion by Japan Esophageal Society (JES) classification and inter- and intraobserver agreement of JES intrapapillary capillary loop (IPCL) classification. Methods: It was a retrospective observational study that has analyzed 136 patients with esophageal malignant neoplasia with magnifying endoscopy narrow band imaging to compare JES’s IPCL classification to the histopathologic findings and to evaluate the inter- and intraobserver agreement. Results: Histopathologic examinations revealed 34 (25.7%) intraepithelial neoplasias, 70 (51.5%) squamous cell carcinomas (SCCs) in the epithelium or with invasion into the lamina propria mucosa, 21 (15.4%) SCCs with invasion into the muscularis mucosa or mild invasion into the submucosa, and 11 (8.1%) SCCs with moderate or deep invasion into the submucosa. IPCL types B1, B2, and B3 also showed high accuracies of 80.8, 83.1, and 94.1%, respectively. The kappa values for inter- and intraobserver agreements of the IPCL classifications were moderate to almost perfect. Conclusions: In the present study, the JES’s IPCL classification has good accuracy to predict the depth of SCC invasion and moderate to almost perfect intra- and interobserver agreements.

A Simple Bioreactor-Based Method to Generate Kidney Organoids fromPluripotent Stem Cells.

SummaryKidney organoids made from pluripotent stem cells have the potential to revolutionize how kidney development, disease, and injury are studied. Current protocols are technically complex, suffer from poor reproducibility, and have high reagent costs that restrict scalability. To overcome some of these issues, we have established a simple, inexpensive, and robust method to grow kidney organoids in bulk from human induced pluripotent stem cells. Our organoids develop tubular structures by day 8 and show optimal tissue morphology at day 14. A comparison with fetal human kidneys suggests that day-14 organoid tissue most closely resembles late capillary loop stage nephrons. We show that deletion of HNF1B, a transcription factor linked to congenital kidney defects, interferes with tubulogenesis, validating our experimental system for studying renal developmental biology. Taken together, our protocol provides a fast, efficient, and cost-effective method for generating large quantities of human fetal kidney tissue, enabling the study of normal and aberrant kidney development.

Retinal Vascular Degeneration in the Transgenic P23H Rat Model of Retinitis Pigmentosa.

Retinitis pigmentosa (RP) is a group of inherited retinal degenerative diseases involving a progressive degeneration of photoreceptor cells. Following the loss of photoreceptors, retinal vascularization tends to decrease, which seems to play a role in the degenerative process of retinal cells. This study reports changes in retinal vascular network architecture in the P23H rat model of RP at different stages of retinal degeneration. Homozygous P23H line-3 rats of ages ranging from 18 days to 16 months were used in this study. Age-matched Sprague-Dawley (SD) rats were used as control animals. Vertical sections and wholemount retinas were immunolabeled for type IV collagen or stained using NADPH diaphorase histochemistry, and retinal vascular networks were drawn using a camera lucida. The superficial and deep capillary plexus (DCP) were fully developed at P18 in P23H rat retinas and showed no differences from the control animals. In 4-month-old P23H rat retinas, the superficial and intermediate capillary plexus were similar to those observed in age-matched SD rats, but a reduction in the DCP could be observed in these animals, with a significant decrease in both capillary density and capillary loops. At 16 months, the DCP was completely lost, and only vessels exhibiting an abnormal, tortuous dead-end could be observed. The middle capillary plexus had virtually disappeared at this age. Only perpendicular vessels connecting the superficial and DCP were found. The superficial plexus showed no changes in the vascular surface with age. In RP, photoreceptor loss is accompanied by degenerative changes in the retinal vascular network. The disruption of the capillary plexus, with loss of capillary density and capillary loops, can hamper the normal supply of oxygen and nutrients to retinal cells, thus accelerating retinal degeneration. Therefore, changes in retinal vascularization must be taken into account in the design of therapies targeting retinal degenerative diseases.

Effects of the differential expression of ZO-1 and ZO-2 on podocyte structure and function.

Glomerular podocytes in the kidney originate from columnar epithelial cells possessing tight junctions. During podocyte differentiation, tight junctions are replaced by slit diaphragms, which are formed between foot processes and function as a blood filtration barrier. Although the expression of most tight junction components is suppressed during podocyte differentiation, several components, including ZO-1 and ZO-2, are consistently expressed. We recently showed that podocyte-specific deletion of ZO-1 gene impaired slit diaphragm formation, leading to proteinuria and glomerular sclerosis. Here, we address the relevance of ZO-2, whose sequence is highly similar to ZO-1, in the maintenance of the structure and function of podocytes. In glomerular development, the spatiotemporal expression of ZO-2 was similar to that of ZO-1 until the capillary loop stage. Subsequently, the distribution patterns of ZO-1 and ZO-2 diverged at the maturation stage, when slit diaphragms are formed. This divergence could partly rely on the ability of ZO-2 to interact with the slit diaphragm membrane proteins. Podocyte-specific deletion of the ZO-2 gene did not cause overt defects; however, double knockout of ZO-1 and ZO-2 genes accelerated the defects observed in ZO-1 knockout mice. These results suggest that ZO-2 plays supportive roles in the ZO-1-dependent regulation of podocyte filtration barrier.

Can the depth of invasion of early esophageal cancer be predicted based on endoscopic evidence?

Esophageal cancer is one of the leading causes of cancer-related death worldwide. Its poor prognosis is related to an often late diagnosis. An earlier diagnosis and treatment however, is related to a better outcome. Early stage esophageal cancer can be diagnosed and treated endoscopically with minimally invasive techniques, which is associated with lower mortality and morbidity than surgery. Whether esophageal carcinoma can be treated endoscopically depends mainly on the risk of lymph node metastasis, which itself correlates to the invasion depth of the tumor. The question is whether endoscopy can accurately determine the invasion depth and thus the treatment modality. Articles used for this review were identified by searches of PubMed and references of relevant articles. Lesion morphology has some predictive value for the depth of invasion for squamous cell carcinoma (SCC) and esophageal adenocarcinoma (EAC). An intramucosal cancer generally has a flat appearance (Paris 0-IIa, 0-IIb,). By contrast, a submucosally invasive cancer often has an excavated (0-IIc, 0-III) and sometimes a polypoid morphology (0-I). In SCC, classification of surface vessels and intrapapillary capillary loops (IPCLs) allows accurate assessment of invasion depth. Generally, mucosal lesions are an indication for endoscopic treatment. However recent studies have shown that tumors with submucosal infiltration and low risk profile for metastasis can also be treated safely by endoscopic resection. Endoscopic assessment allows a rather accurate estimation of invasion depth of early esophageal cancer. To determine the final treatment modality however the final histological staging obtained by endoscopic mucosal resection (EMR) or endoscopic submucosal dissection (ESD) is crucial.

Narrow band imaging for risk stratification of glottic cancer within leukoplakia.

This study investigates relevance of narrow band imaging (NBI) in stratifying risk of malignant transformation within leukoplakia. We conducted a prospective analysis that included 62 patients with 91 changes of leukoplakia on vocal folds. The NBI was obtained before microsurgery. Categorization of the lesion as benign was made when vessels of surrounding epithelium were classified as type I, II, or IV according to Ni classification. If there were visualized intraepithelial papillary capillary loops of type V, the lesion was classified as malignant. Results were compared to the histopathological diagnosis. The NBI assessment classified 75 lesions (82.4%) as benign and 16 (17.6%) as malignant. Histopathological results revealed the diagnosis of no dysplastic changes or low grade dysplasia in 77 cases (84.6%). Another 14 cases (15.4%) occurred with high-grade dysplasia, carcinoma in situ, and invasive cancer. Sensitivity, specificity, and accuracy of NBI in predicting malignancy within leukoplakia were 100%, 97.4%, and 97.8%, respectively. The kappa index was 0.92 (95% confidence interval 81.1%-100%). The noninvasive procedure of an NBI endoscopy may be recommended as an accurate method in predicting the risk of malignant transformation within the vocal fold leukoplakia and, therefore, would be useful in the clinic for planning the patient's therapy.

Young Ossabaw Pigs Fed a Western Diet Exhibit Early Signs of Diabetic Retinopathy.

PurposeRecent clinical data suggest an increasing prevalence of obesity and type 2 diabetes in adolescents, placing them at high risk of developing diabetic retinopathy during adult working years. The present study was designed to characterize the early retinal and microvascular alterations in young Ossabaw pigs fed a Western diet, described as a model of metabolic syndrome genetically predisposed to type 2 diabetes.MethodsFour-month-old Ossabaw miniature pigs were divided into two groups, lean and diet-induced obesity. Obese pigs were fed a Western diet with high-fat/high-fructose corn syrup/high-choleric content for 10 weeks. Blood and retina were collected for biochemical profiling, trypsin digest, flatmounts, Fluoro-Jade C staining, electron microscopy, quantitative PCR, immunohistochemistry, and Western blots.ResultsYoung Ossabaw pigs had elevated fasting blood glucose after feeding on a Western diet for 10 weeks. Their retina showed disrupted cellular architecture across neural layers, with numerous large vacuoles seen in cell bodies of the inner nuclear layer. Microvessels in the obese animals exhibited thickened basement membrane, along with pericyte ghosts and acellular capillaries. The pericyte to endothelial ratio decreased significantly. Retina flatmounts from obese pigs displayed reduced capillary density, numerous terminal capillary loops, and string vessels, which stained collagen IV but not isolectin IB4. Quantitative PCR and Western blots showed significantly high levels of basement membrane proteins collagen IV and fibronectin in obese pigs.ConclusionsThis is the first study to describe the ultrastructural neuronal and vascular changes in the retina of young Ossabaw pigs fed a Western diet, simulating early signs of diabetic retinopathy pathogenesis.

The microvascular and morphostructural changes of nails in psoriatic patients with nail disease; a link between ultrasound and videocapillaroscopy findings in the nailfold.

The objective of this study is to evaluate the link between nail fold vessel resistive index (NVRI) measured by ultrasound (US) and capillary loops diameters measured using nailfold videocapillarascopy (NVC), and to assess the morphological appearance of the nail bed in patients with psoriatic nail disease (PND) as compared with healthy controls (HCs). This study was conducted in patients with PND and HCs. General demographic data were collected and clinical assessments were performed for all subjects. The nail plate thickness (NPT) was measured on gray scale using US. The NVRI was measured using color Doppler (CD) US. The measurements of the apical, arterial, venous limb diameters and morpho-structural changes (tortuous, cross-linked capillaries) were assessed using NVC. Thirty-four patients with PND and 15 HCs were enrolled in this study. The two groups were matched for age and body mass index (BMI). Patients with PND had higher NPT and NVRI in comparison with HCs [(20 (17-23) vs 14 (14-15), p<0.001), (0.55 (0.51-0.61) vs 0.43 (0.38-0.49), p<0.001), respectively]. A higher proportion of patients with PND had tortuous capillaries than HCs (62% and 20% respectively, p=0.005). The mean NVRI was higher in patients with PND who had tortuous capillaries than patients who did not have tortuous capillaries (0.58 (0.7) and 0.52 (0.09), respectively p=0.033). Microvascular changes can be detected easily using non-invasive methods such as US and NVC. These methods can provide an objective data to better assess PND.

Prolonged cold ischemia storage of renal allografts shifts DSA specificity and enhances glomerular injury.

Abstract Prolonged cold ischemia storage (CIS) of donor organs is a major risk factor for allograft injury. We tested how increased CIS time influences humoral alloimmunity in a mouse model of renal transplantation. B6 (H-2b) mice received BALB/c (H-2d) renal allografts subjected to 0.5 h or 6 h CIS. At 14 d posttransplant, 6 h CIS recipients had increased titers of donor-MHC class II but not class I reactive IgG DSA, elevated frequencies of both class I and class II-reactive antibody secreting cells, higher frequencies of donor-reactive IFNγ-secreting T cells and higher proportion of graft glomeruli infiltrated with macrophages compared to 0.5 h CIS controls. B cell depletion inhibited DSA generation, intragraft C4d deposition and macrophage glomerular infiltration on d14. Regardless of CIS duration, serum creatinine levels at d 60 posttransplant were comparable to non-transplanted mice. By d 60, 0.5 h CIS grafts had moderate segmental sclerosis in a limited number of glomeruli. In contrast, 6 h CIS grafts had extensive glomerular injury including thickened capillary loops, sclerosis, thrombotic microangiopathy, intracapillary fibrin thrombi, and mesangiolysis, and elevated intragraft levels of acute kidney injury markers NGAL and osteopontin. In addition, 6 h CIS recipients had increased serum levels of anti-class II but not anti-class I IgG DSA whereas the frequencies of IFNγ-producing donor-reactive T cells were low and comparable in both groups. Our findings suggest that the augmented humoral rather than cellular immune responses account for the transplant glomerulopathy after prolonged CIS. Posttransplant inflammation spreads the specificity of DSA responses from class I to class II and may thus influence renal allograft pathology.

Nailfold Capillaroscopy of Fingers and Toes - Variations of Normal.

Nailfold capillaroscopy is the only method for morphological assessment of nutritive capillaries. The literature data about capillaroscopic findings in healthy individuals are scarce. To evaluate and compare the capillaroscopic findings of fingers and toes in healthy subjects. 22 healthy individuals were included in the study. Capillaroscopic examination was performed with videocapillaroscope Videocap 3.0 (DS Medica). Exclusion criteria were as follows: history of vasospasm, presence of accompanying diseases, taking any medications, arterial hypertension in first degree relatives, overweight or obesity (body mass index > 25kg/m2) and presence of chronic arterial or venous insufficiency. Poor visibility of nailfold capillaries was found significantly more frequently in the toes (22.7%, 5/22) as compared with fingers (0/22). Slight irregularities in capillary distribution and orientation to their parallel axis were significantly more common in the toes (31.8%, 7/22) as compared with fingers (9%, 2/22), (p<0.05). The mean diameter of the arterial (0.012±0.002mm) and the venous limb (0.017±0.002mm) of the toes did not differ significantly as compared to the respective parameters in the fingers (0.013±0.002mm for the arterial limb, p=0.46 and 0.018±0.002mm for the venous limb, p=0.25). The mean capillary density also did not differ significantly in the fingers and toes. The mean capillary length of the toes (0.165±0.096mm) was shorter as compared with hands (0.220±0.079mm), but the difference was not statistically significant (p=0.37). Presence of tortuous capillaries (>10%) was found significantly more often in the toes (12/22) as compared with fingers (6/22, χ2=6.769, p<0.05). Short capillary loops (length<100µm) were observed significantly more often in the toes (11/22 - toes, 1/22 - fingers, χ2=14.666, p<0.05). Capillaroscopic examination of the toes shows some differences as compared to those of the fingers such as greater number of cases with poor visibility and slight irregularities of distribution, greater number of shorter capillaries and increased tortuosity, which might be related to the thicker epidermis of the toes and increased capillary pressure due to gravity. The values of the major capillaroscopic parameters such as capillary diameters and capillary density in fingers and toes do not differ significantly.

Capillaroscopic Findings in Primary Fibromyalgia.

Although Raynaud's Phenomenon (RP) is observed in a significant proportion of patients with primary fibromyalgia, the available data on capillaroscopic findings in primary fibromyalgia are scarce. The purpose of the study was to assess the capillaroscopic pattern in patients with primary fibromyalgia. 26 patients with primary fibromyalgia (25 women and 1 man) were included in the study. Mean age was 55±10 years. As control groups were examined 31 patients with primary RP and 35 healthy volunteers. Capillaroscopic examination was performed with a videocapillaroscope Videocap 3.0 (DS Medica), magnification 200x with analysis of the main capillaroscopic parameters as follows: capillary distribution, shape, mean capillary diameters and length, mean capillary density, visibility of the subpapillary plexus; presence of avascular areas, microhaemorrhages and neoangiogenic capillaries. Symptoms of RP were observed in 65% (17/26) of the cases with primary fibromyalgia. At capillaroscopic examination, the most frequent finding in patients with primary fibromyalgia was the presence of capillary dilation in 85% (22/26) of the patients - both in cases with and without RP. However, the mean arterial and venous capillary diameters were significantly higher in the subgroup of fibromyalgia patients with clinical symptoms of RP. Of note, microvascular abnormalities characteristic of connective tissue diseases could not be observed in primary fibromyalgia patients. Analogous changes - presence of dilated capillaries - were found in 96.6% (29/30) of patients with primary RP. In our study, the most frequent capillaroscopic finding in patients with primary fibromyalgia was the presence of dilated capillary loops analogous to primary RP. Capillaroscopic signs suggestive of connective tissue disease could not be found in primary fibromyalgia patients.

Evaluation of a Miniaturized Biologically Vascularized Scaffold in vitro and in vivo

In tissue engineering, the generation and functional maintenance of dense voluminous tissues is mainly restricted due to insufficient nutrient supply. Larger three-dimensional constructs, which exceed the nutrient diffusion limit become necrotic and/or apoptotic in long-term culture if not provided with an appropriate vascularization. Here, we established protocols for the generation of a pre-vascularized biological scaffold with intact arterio-venous capillary loops from rat intestine, which is decellularized under preservation of the feeding and draining vascular tree. Vessel integrity was proven by marker expression, media/blood reflow and endothelial LDL uptake. In vitro maintenance persisted up to 7 weeks in a bioreactor system allowing a stepwise reconstruction of fully vascularized human tissues and successful in vivo implantation for up to 4 weeks, although with time-dependent decrease of cell viability. The vascularization of the construct lead to a 1.5× increase in cellular drug release compared to a conventional static culture in vitro. For the first time, we performed proof-of-concept studies demonstrating that 3D tissues can be maintained within a miniaturized vascularized scaffold in vitro and successfully implanted after re-anastomosis to the intrinsic blood circulation in vivo. We hypothesize that this technology could serve as a powerful platform technology in tissue engineering and regenerative medicine.