Comparison of the prevalence of human papilloma virus infection in histopathologically confirmed premalignant oral lesions and healthy oral mucosa by brush smear detection.

OBJECTIVE: The role of human papilloma virus (HPV) infections in oral carcinogenesis is an important topic of research in maxillofacial oncology. Nevertheless, the association between such infections in the oral cavity and the development of oral precancerous lesions remains unclear. The aim of this study was to evaluate the association between oral HPV infections and oral leukoplakia or erythroplakia. STUDY DESIGN: The case control study included 118 patients with manifest oral leukoplakia or erythroplakia, who underwent surgical biopsy, including a histopathologic grading of the lesion, and 100 control patients without any oral lesions. HPV detection was achieved with a noninvasive brush smear method (Digene Cervical Sampler, Hybrid Capture II-Test). Logistic regression analysis was performed to assess the associations. RESULTS: A significant association was found between high-risk oral HPV infection and the presence of oral premalignant lesions (P = .001). Among all other evaluated parameters, only smoking showed a significant association with the presence of oral lesions. CONCLUSIONS: Oral HPV infections may play a role in the pathogenesis of premalignant oral lesions.

Isolation and characterization of CD133+CD34+VEGFR-2+CD45- fetal endothelial cells from human term placenta.

The phenotypes and functions of endothelial cells (EC), a heterogeneous cell population, vary along the vascular tree and even in the same organ between different vessels. The placenta is an organ with abundant vessels. To enhance further knowledge concerning placenta derived EC, we develop a new method for isolation, purification and culture of these EC. Moreover, in order to investigate the peculiarity of placenta derived EC we compare their phenotypic and functional characteristics with human dermal lymphatic endothelial cells (HDLEC) and human umbilical vein endothelial cells (HUVEC). Freshly isolated placenta derived EC displayed an elongated shape with pale cytoplasm and showed the typical cobblestone pattern of EC but also a swirling pattern when confluent. FISH-analyses of the isolated EC from placentae of male fetus revealed an XY genotype strongly indicating their fetal origin. Characterisation of placenta derived fetal EC (fEC) underlined their blood vessel phenotype by the expression of vWF, Ulex europaeus lectin-1, HLA-class I molecules, CD31, CD34, CD36, CD51/61, CD54, CD62E, CD105, CD106, CD133, CD141, CD143, CD144, CD146, VEGFR-1, VEGFR-2, EN-4, PAL-E, BMA120, Tie-1, Tie-2 and α-Tubulin. In contrast to previous reports the expression of lymphatic markers, like VEGFR-3, LYVE-1, Prox-1 and Podoplanin was consistently negative. Haematopoietic surface markers like CD45 and CD14 were also always negative. Various functional tests (Dil-Ac-LDL uptake, Matrigel assay and TNF-α induced upregulation of CD62E and CD54) substantiated the endothelial nature of propagated fEC. At the ultrastructural level, fEC harboured numerous microvilli, micropinocytic vesicles at their basis, were rich in intermediate filaments and possessed typical Weibel - Palade bodies. In conclusion, the placenta is a plentiful source of fetal, microvascular, blood EC with an expression profile (CD34+, CD133+, VEGFR-2+, CD45-) suggestive of an endothelial progenitor phenotype.

Imaging of placental transport mechanisms: a review.

Functional analysis of material transfers requires precise statement of residence times in each tissue compartment. For the placenta, neither extractive biochemistry, isotope partitioning, nor mass-based quantitative assays provide adequate spatial resolution to allow the necessary precision. Dual-perfusion assays of material transfer in isolated placental cotyledons provide time-series data for two compartments, the maternal and fetal blood, but fail to distinguish the two cellular compartments (syncytiotrophoblast, fetal endothelium) which actively regulate rates of transfer in each direction for essentially every important molecule type. At present, no definitive technology exists for functional analysis of placental transfer functions. The challenge in developing such a technology lies in the exquisitely small and delicate structures involved, which are scaled at cellular and subcellular sizes (between 50 nm and 50 microm). The only available technologies attaining this high spatial resolution are imaging technologies, primarily light and electron microscopy. To achieve the high-quality images necessary, confocal laser scanning microscopy (CLSM) is required, to provide a uniform optical sectioning plane. In turn, this requires relatively high fluorescence intensities. Design of an adequate technology therefore bases on CLSM imaging fluorochrome-tagged tracers. The temporal resolution necessary to analyse placental material transfers is expected to be of the order of a few seconds, so that conventional wet-fixation protocols are too slow. For adequately rapid fixation, snap-freezing is required. As part of this review we report results obtained from an appropriately designed experimental protocol, analysed by CLSM and transmission electron microscopy (TEM). The images acquired were tested for uniformity of illumination and fluorescence emission strength. Relevant data was encoded in the green channel of the trichrome images obtained, and this was thresholded by application of strict quantitative criteria. The thresholding procedure is suitable for automation and produces reproducible, objectifiable results. Thresholded images were subjected to image calculation procedures designed to highlight image elements (pixels) containing (green) fluorescence associated with the tracer protein; all other sources of fluorescence were visualised in the final images only if no green fluorescence was detectable in that pixel. The resulting images were maps, showing the distribution of tracer molecules at a predefined time interval after perfusion of the tracer into the vital (term) cotyledon. Spatial resolution was routinely better than 1 microm and temporal resolution was approximately 5s. At timepoints up to 10 min after intravital application into the fetal vascular circulation, tracer was associated with capillaries in the villous structures, and no tracer was observed in the syncytiotrophoblast. Clear distinction was achieved between the four tissue compartments relevant to placental transfers, thus providing a novel technology capable of generating high-quality data concerning the regulation of transfers of any molecule that can bear a fluorescent tag. The potential applications of this methodology lie in analyses of factors influencing the rates of fetomaternal and maternofetal exchanges (for example, drugs), and of functional responses of the placental regulation to pathophysiological conditions such as hypoxia.

Measurement of tryptophan, kynurenine and neopterin in women with and without postpartum blues.

BACKGROUND: Activation of the tryptophan-degrading enzyme indoleamine (2,3)-dioxygenase was demonstrated to be critically involved in tolerance induction to prevent fetal rejection. Our study was designed to examine alterations of tryptophan and its catabolic product kynurenine in the postpartum period and to compare them to neopterin as an immunological marker. METHODS: 95 healthy women delivering without complications provided blood during labour, and 2 and 4 days after birth. The blood samples were analysed for concentrations of tryptophan, kynurenine and neopterin. Women were asked to perform the Edinburgh Postnatal Depression Scale (EPDS) on days 2 and 4. RESULTS: In women without blues symptoms (n=86) tryptophan concentrations increased within 2 days after birth, whereas they did not change in women with postpartum blues (n=9; 9.5%). The group difference reached statistical significance (p<0.05). The change of the kynurenine to tryptophan ratio (kyn/trp), which estimates the degree of tryptophan degradation, was also different between the two groups at days 0 and 2 (p<0.05). Neopterin concentrations decreased between days 2 and 4 (p<0.05), but there were no differences between the two groups. LIMITATIONS: Our study population had a low prevalence of postpartum blues symptoms. CONCLUSION: Low postpartal mood is associated with continuously low serum tryptophan after delivery due to an increased degradation to kynurenine, but is independent of the postpartal course of neopterin.

Design, synthesis, physical and chemical characterisation, and biological interactions of lectin-targeted latex nanoparticles bearing Gd-DTPA chelates: an exploration of magnetic resonance molecular imaging (MRMI).

The physical and chemical parameters involved in the design and synthesis of biospecifically targeted nanoparticulate contrast media for magnetic resonance molecular imaging (MRMI) were explored in this pilot investigation. Latex nanoparticles 100, 400 and 900 nm in diameter were doubly derivatised, first with tomato lectin and then with gadolinium(III)-diethylenetriamine pentaacetic acid (Gd-chelates) to target them to epithelial and endothelial glycocalyceal N-glycans and to generate contrast enhancement in magnetic resonance imaging (MRI). After intravenous injection into mice, human placental cotyledons or human Vena saphena magna, contrasty images of the vascular structures were obtained in 1.5 T MRI with spatial resolution 0.1 mm in the imaging plane and 0.6 mm in the z axis, persisting >60 min and resistant to washing out by buffer rinses. Ultrastructural analysis of the nanoparticles revealed the targeting groups at the nanoparticle surfaces and the distribution of the Gd-chelates within the nanoparticles and enabled counts for use in determining relaxivity. The relaxivity values revealed were extremely high, accounting for the strong MR signals observed. Occasionally, nanoparticles larger than 100 nm were seen in close spatial association with disrupted regions of cell membrane or of collagen fibrils in the extracellular matrix. The data suggest that 100-nm nanoparticles generate adequate contrast for MRMI and cause least disruption to endothelial cell surfaces.

Chronic pain syndromes and their relation to childhood abuse and stressful life events.

OBJECTIVE: Childhood abuse, stressful life events, and depression have been repeatedly reported to correlate with chronic pain, but little is known about the mutual relationships among these variables. METHODS: Forty-three women with chronic pelvic pain (CPP), 40 female patients with chronic low-back pain (CLBP), and a female pain-free control group (n=22) were investigated by means of a semistructured interview assessing childhood sexual and physical abuse as well as stressful life events. Additionally, the Beck Depression Inventory (BDI) was used. For multivariate analyses, structured equation modeling was applied. RESULTS: Childhood physical abuse, stressful life events, and depression had a significant impact on the occurrence of chronic pain in general, whereas childhood sexual abuse was correlated with CPP only. Moreover, childhood sexual abuse was related to depression. Both childhood sexual and physical abuse showed a close relationship to an increased occurrence of stressful life events. CONCLUSION: There are complex mutual interactions among childhood abuse, stressful life events, depression, and the occurrence of chronic pain. Therefore, clinicians should take into consideration these psychosocial factors while treating chronic pain patients.

Perinatal bone turnover in term human neonates and the influence of maternal smoking.

Bone turnover in neonates appears independently of the comparably low maternal bone turnover, but there is only sparse information on the effect of the in utero environment on fetal bone turnover. Postnatally, the resuming growth velocity and alterations in mineral homeostasis affect neonatal bone turnover. This study evaluated the relationship of bone marker concentrations to maternal and fetal auxological variables as well as maternal smoking and assessed the short-term change in bone markers during the first days of life. Serum markers of bone formation [osteocalcin and bone-specific alkaline phosphatase (BALP)] and bone resorption (C-terminal telopeptide of type I collagen) were measured in cord blood and at discharge (median d 3) in 69 healthy term neonates. Concentrations of BALP were significantly lower in neonates of smokers (n = 16) compared with nonsmokers (n = 53), both at birth (p = 0.013) and at discharge (p = 0.036). Both cord osteocalcin and BALP were negatively related to maternal weight and maternal body mass index. Maternal smoking and pregnancy weight gain were the predictors of cord BALP (r2 = 0.24; p < 0.001), whereas the mode of delivery best predicted cord C-terminal telopeptide of type I collagen levels (r2 = 0.19; p < 0.001). C-terminal telopeptide of type I collagen and osteocalcin increased significantly (p < 0.001) from birth to discharge, whereas BALP levels did not change significantly during the same period. Our results suggest that maternal smoking during pregnancy and maternal obesity may have a negative impact on fetal bone formation. The significant increase of osteocalcin and C-terminal telopeptide of type I collagen may result either from an increase in bone turnover or altered renal clearance.