Micronuclei and nuclear anomalies in urinary exfoliated cells of subjects in radionuclide-contaminated regions

¹³⁷Cs contamination in living or agricultural environments may contribute to significant human internal exposure and cause adverse health effects. Contamination by ¹³⁷Cs and other radionuclides was detected in a river valley in northern Taiwan, in the 1990s. Given that the radioactivity appeared to be widely distributed in soil, rice and several other food plants in the areas surrounding several communities in the late 1990s [Y.B. Nabyvanents, T.F. Gesell, M.H. Jen, W.P. Chang, Distribution of ¹³⁷Cs in soil along Ta-han River Valley in Tau-Yuan County in Taiwan, J. Environ. Radioact. 54 (2001) 391], its possible impact on local occupants was further studied.Ten subjects in three families residing continuously in the highly contaminated valley and 10 non-exposed subjects matched for age, sex, and cigarette smoking habits from neighboring communities were evaluated for micronucleus frequencies and for degenerative nuclear changes in urinary exfoliated epithelial cells (EE cells). Micronucleus frequencies (‰) were significantly higher in the exposed subjects (4.79±1.21‰) than in the reference subjects (2.73±0.59‰; Wilcoxon 2-sample test, P value 0.0004). There were also higher frequencies of EE cells with karyolysis and condensed chromatin in the exposed subjects than in reference subjects. These results indicate that genotoxic and/or mutagenic effects on urinary epithelial cells occur in human subjects who have resided for a long time in a radioactively contaminated environment.     

Patterns of age-specific mortality and molding of senescence by natural selection derived via the life-history regression model

An individual based life-history regression setup is introduced not only as an alternative to the ‘reproductive effort model’ in life-history theory, but as a new platform on which the nature of reproductive costs can be explicitly determined and tested under different demographic environmental conditions. Distinctively this regression model is composed of two age-specific features: one is an estimable baseline mortality rate describing the life-history of a population hypothetically undertaking no reproduction, but investing all vital resources into somatic maintenance and growth; the other is a time-dependent covariate encoding dynamic impacts incurred from individual's schedule of reproduction. Regression parameters embedded in the time-dependent covariate explicitly stand for various effects of reproductive costs on future survival relative to the standard described by the baseline mortality rate. Consequently the age-specific mortality is in a compositional structure and gives rise to a wide spectrum of well known mortality curves. Also this compositional structure renders molding forces on senescence by natural selection crucially dependent on patterns of the schedule of reproduction. All numerical evaluations of patterns are performed with two distinct reproductive schedules to illustrate the essential differences from classic results in literature of evolution of life-history.     

Soil-productivity relationships and organic matter turnover in dry tropical forests of the Florida Keys

Soils and aboveground production in five types of upland forest in the Florida Keys were examined. Throughout the habitat gradient represented by these forest types, the soils were predominantly shallow and organic, forming in place directly on the limestone bedrock. However, the well-drained soils in the most productive broadleaved forests were deep enough to qualify as Histosols (Folists). Soils decreased in electrical conductivity and increased in nutrient content with increasing aboveground production. At 3–12 Mg ha^–1 yr^–1, production was within the range reported for dry tropical forests. Measured rates of decomposition were moderate or fast, and estimates of the organic C turnover of several soils based on their bomb radiocarbon signature were 100 years or less. In the face of these rapid turnover rates, we attribute the development of organic soils to the absence of mineral residues from weathering of the underlying limestone bedrock. Fast turnover of organic matter, and rapid and efficient cycling of nutrients are necessary to sustain the high production rates obtained on these shallow organic soils.     

TSG101 expression in gynecological tumors: relationship to cyclin D1, cyclin E, p53 and p16 proteins.

Recent studies have shown that in vitro steady-state expression of the tumor susceptibility gene TSG101 is important for maintenance of genomic stability and cell cycle regulation. To determine the contribution of TSG101 expression in neoplastic formation, expression of TSG101 protein levels were evaluated in primary ovarian and endometrial adenocarcinoma tumors. Expression of TSG101 was also examined in various tumor cell lines (PA-1, AN3CA, HeLa, HS578T, HCT116). Full-length TSG101 protein was detected in these tumors and cell lines indicating that intragenic deletions were not characteristic of TSG101. In addition, TSG101 protein levels were compared with aberrations of prominent cell cycle regulatory molecules such as cyclin D1, cyclin E, p16 and p53. Reduced TSG101 protein was observed in 36% (8/22) of ovarian and 17% (1/6) of endometrial adenocarcinoma. Aberrant levels of p53, p16, cyclin D or E were comparable to published studies indicating that the clinicopathological distribution of these cases did not favor advanced stage tumors. Altogether, these findings suggest that a down-regulation of TSG101 is associated with tumorigenesis in a subgroup of gynecological tumors.     

Impaired Abdominal Wall Development and Deficient Wound Healing in Mice Lacking Aortic Carboxypeptidase-Like Protein

Aortic carboxypeptidase-like protein (ACLP) is a member of a diverse group of proteins that contain a domain with similarity to that of the Dictyostelium discoideum protein discoidin I. The discoidin domain has been identified in mammalian milk fat globule membrane proteins, blood coagulation factors, and receptor tyrosine kinases, where it may facilitate cell aggregation, adhesion, or cell-cell recognition. Here we show that ACLP is a secreted protein that associates with the extracellular matrix (ECM). During mouse embryogenesis, ACLP is abundantly expressed in the ECM of collagen-rich tissues, including the vasculature, dermis, and the developing skeleton. We deleted the ACLP gene in mice by homologous recombination. The majority of ACLP(-/-) mice die perinatally due to gastroschisis, a severe disruption of the anterior abdominal wall and herniation of the abdominal organs. ACLP(-/-) mice that survived to adulthood developed nonhealing skin wounds. Following injury by a dermal punch biopsy, ACLP(-/-) mice exhibited deficient wound healing compared with controls. In addition, dermal fibroblasts isolated from ACLP(-/-) 18.5-day-postconception embryos exhibited a reduced proliferative capacity compared with wild-type cells. These results indicate that ACLP is an ECM protein that is essential for embryonic development and dermal wound healing processes.     

Resveratrol inhibits copper ion-induced and azo compound-initiated oxidative modification of human low density lipoprotein.

To investigate whether resveratrol, a polyphenolic compound in red wine, affects the oxidation of human low density lipoprotein (LDL), LDL purified from normolipidemic subjects was subjected to Cu(2+)-induce and azo compound-initiated oxidative modification, with and without the addition of varying concentrations of resveratrol. Modification of LDL was assessed by the formation of thiobarbituric acid reactive substances (TBARS) and changes in the relative electrophoretic mobility (REM) of LDL on agarose gels. Resveratrol (50 microM) reduced TBARS and REM of LDL during Cu(2+)-induced oxidation by 70.5% and 42.3%, respectively (p < 0.01), and prolonged the lag phase associated with the oxidative modification of LDL by copper ion or azo compound. These in vitro results suggest that resveratrol may afford protection of LDL against oxidative damage resulting from exposure to various environmental challenges, possibly by acting as a free radical scavenger.     

Inexpensive, semi-automated system for measuring mechanical properties of soft tissues.

Stiffness and strength are important properties of many tissues, but standard material-testing equipment is expensive, often ill-suited for testing soft tissues, and rarely accessible to biologists. We describe a system built around a microcomputer and an electronic balance which is particularly well-suited for measuring stress and strain in small samples of soft tissue. We use a discarded floppy disk drive as a linear actuator to strain the sample, while an electronic balance measures the tension (used to calculate stress). We give an algorithm for a program to drive a microcomputer which controls the floppy disk drive via its parallel port and records the balance measurements via its serial port. We used this system to obtain stress-strain curves from a sample of latex rubber and a sample of soft insect cuticle. Three tests of the rubber sample gave nearly identical results, with smooth, J-shaped stress-strain curves. The stress-strain curves gave a modulus elasticity value of 1.72 Mpa over the steep, straight region, well within the range for natural latex rubber. We also tested a sample of abdominal cuticle from a caterpillar (Manduca sexta). The caterpillar cuticle had a J-shaped stress-strain curve with a modulus of elasticity of 2.11 Mpa over the steep part of the curve. J. Exp. Zool. 284:374-378, 1999.     

Aberrant stromal tissue factor localisation and mycolactone-driven vascular dysfunction,exacerbated by IL-1β, are linked to fibrin formation in Buruli ulcer lesions

Buruli ulcer (BU) is a neglected tropical disease caused by subcutaneous infection with Mycobacterium ulcerans and its exotoxin mycolactone. BU displays coagulative necrosis and widespread fibrin deposition in affected skin tissues. Despite this, the role of the vasculature in BU pathogenesis remains almost completely unexplored. We hypothesise that fibrin-driven ischemia can be an ‘indirect’ route to mycolactone-dependent tissue necrosis by a mechanism involving vascular dysfunction. Here, we tracked >900 vessels within contiguous tissue sections from eight BU patient biopsies. Our aim was to evaluate their vascular and coagulation biomarker phenotype and explore potential links to fibrin deposition. We also integrated this with our understanding of mycolactone’s mechanism of action at Sec61 and its impact on proteins involved in maintaining normal vascular function. Our findings showed that endothelial cell dysfunction is common in skin tissue adjacent to necrotic regions. There was little evidence of primary haemostasis, perhaps due to mycolactone-dependent depletion of endothelial von Willebrand factor. Instead, fibrin staining appeared to be linked to the extrinsic pathway activator, tissue factor (TF). There was significantly greater than expected fibrin staining around vessels that had TF staining within the stroma, and this correlated with the distance it extended from the vessel basement membrane. TF-induced fibrin deposition in these locations would require plasma proteins outside of vessels, therefore we investigated whether mycolactone could increase vascular permeability in vitro. This was indeed the case, and leakage was further exacerbated by IL-1β. Mycolactone caused the loss of endothelial adherens and tight junctions by the depletion of VE-cadherin, TIE-1, TIE-2 and JAM-C; all Sec61-dependent proteins. Taken together, our findings suggest that both vascular and lymphatic vessels in BU lesions become “leaky” during infection, due to the unique action of mycolactone, allowing TF-containing structures and plasma proteins into skin tissue, ultimately leading to local coagulopathy and tissue ischemia.     

Host tissue proteomics reveal insights into the molecular basis of Schistosoma haematobium-induced bladder pathology

Urogenital schistosomiasis remains a major public health concern worldwide. In response to egg deposition, the host bladder undergoes gross and molecular morphological changes relevant for disease manifestation. However, limited mechanistic studies to date imply that the molecular mechanisms underlying pathology are not well-defined. We leveraged a mouse model of urogenital schistosomiasis to perform for the first time, proteome profiling of the early molecular events that occur in the bladder after exposure to S. haematobium eggs, and to elucidate the protein pathways involved in urogenital schistosomiasis-induced pathology. Purified S. haematobium eggs or control vehicle were microinjected into the bladder walls of mice. Mice were sacrificed seven days post-injection and bladder proteins isolated and processed for proteome profiling using mass spectrometry. We demonstrate that biological processes including carcinogenesis, immune and inflammatory responses, increased protein translation or turnover, oxidative stress responses, reduced cell adhesion and epithelial barrier integrity, and increased glucose metabolism were significantly enriched in S. haematobium infection. S. haematobium egg deposition in the bladder results in significant changes in proteins and pathways that play a role in pathology. Our findings highlight the potential bladder protein indicators for host-parasite interplay and provide new insights into the complex dynamics of pathology and characteristic bladder tissue changes in urogenital schistosomiasis. The findings will be relevant for development of improved interventions for disease control.