Specific characteristics of urethral strictures in a developing country (Brazil).

PURPOSE: Evaluate the main etiologies and clinical characteristics of male urethral stricture disease (USD) in Brazil. METHODS: This multicentric study was performed using retrospective data collected from six Brazilian referral centers of urethral reconstruction. The database comprised data from 899 patients with USD who had undergone surgical treatment from 2008 to 2018. Age, stricture site and primary stricture etiology were identified for each patient. RESULTS: The mean age was 52.13 ± 16.9 years. The most common etiology was iatrogenic (43.4%), followed by idiopathic (21.7%), trauma (21.5%) and inflammatory (13.7%). Of the iatrogenic causes, 59% were secondary to urethral instrumentation (60% by urethral catheterization and 40% by transurethral procedures), 24.8% by other procedures (prostatectomy, radiotherapy, postectomy) and 16.2% by failed hypospadia repairs. Pelvic fracture urethral distraction injuries were responsible for most of the trauma-related strictures (62.7%). When stratified by age, the most common stricture etiology was trauma in the 0-39 years old group (42.8%), idiopathic in the 40-59 years old group (32.4%) and iatrogenic in patients over 60 years old (68%). In regard to the stricture site, 80% presented with an anterior urethral stricture and 20% with a posterior stenosis. In the anterior stenosis group, the most common stricture site was bulbar (39.5%). CONCLUSION: In Brazil, as in many developed countries, the most common cause of urethral stricture diseases is iatrogenic, especially urethral catheterization. These findings emphasize the need of a careful urethral manipulation and a better training of healthcare professionals. Trauma is still responsible for a great proportion of strictures and inflammatory etiologies are now less frequently observed.

[Burn-out, commitment, personality and experiences during work and training; survey among psychiatry residents]. / Bevlogenheid en burn-out in relatie tot persoonlijkheid en context bij aiossen psychiatrie: een verkennend onderzoek.

BACKGROUND: In the last few years international studies have reported on increase in burn-out and depressive symptoms among psychiatry residents. In the field of research, however, commitment and dedication are now being mentioned more frequently as positive factors that counterbalance burn-out. AIM: To find out how a group of Dutch psychiatry residents feel about their work, to discover their degree of burn-out and commitment and to clarify the various factors involved. METHOD: 59 psychiatry residents from four teaching hospitals were asked to complete questionnaires concerning burn-out (U-BOS-C), commitment (UWES-15) and personality (BFI-NL). Respondents were also asked to describe how they felt about their experiences during their work and to give their views on the instruction and training they were receiving. RESULTS: In the U-BOS-C section only four trainees (almost 7%) met the criteria for burn-out. In the BFI-NL section the psychiatry residents obtained significantly lower scores on neuroticism and higher scores on empathy than did a comparable norm group of a similar age. The scores of the psychiatry residents indicated that the term 'being proud of your work' was significantly related to a feeling of commitment and particularly to all subscales that reflected commitment. CONCLUSION: In our study the percentage of psychiatry residents with burn-out is significantly lower than the percentage reported elsewhere in the literature. In fact, our results demonstrate that the psychiatry residents who were the subject of our study regarded themselves as being emotionally stable, friendly and committed to their work.

Ischemia-Reperfusion Injury Enhances Lymphatic Endothelial VEGFR3 and Rejection in Cardiac Allografts.

Organ damage and innate immunity during heart transplantation may evoke adaptive immunity with serious consequences. Because lymphatic vessels bridge innate and adaptive immunity, they are critical in immune surveillance; however, their role in ischemia-reperfusion injury (IRI) in allotransplantation remains unknown. We investigated whether the lymphangiogenic VEGF-C/VEGFR3 pathway during cardiac allograft IRI regulates organ damage and subsequent interplay between innate and adaptive immunity. We found that cardiac allograft IRI, within hours, increased graft VEGF-C expression and lymphatic vessel activation in the form of increased lymphatic VEGFR3 and adhesion protein expression. Pharmacological VEGF-C/VEGFR3 stimulation resulted in early lymphatic activation and later increase in allograft inflammation. In contrast, pharmacological VEGF-C/VEGFR3 inhibition during cardiac allograft IRI decreased early lymphatic vessel activation with subsequent dampening of acute and chronic rejection. Genetic deletion of VEGFR3 specifically in the lymphatics of the transplanted heart recapitulated the survival effect achieved by pharmacological VEGF-C/VEGFR3 inhibition. Our results suggest that tissue damage rapidly changes lymphatic vessel phenotype, which, in turn, may shape the interplay of innate and adaptive immunity. Importantly, VEGF-C/VEGFR3 inhibition during solid organ transplant IRI could be used as lymphatic-targeted immunomodulatory therapy to prevent acute and chronic rejection.

Removal of rotavirus and adenovirus from artificial ground water using hydrochar derived from sewage sludge.

AIMS: To determine the pathogenic virus removal performance of an adsorbent produced from hydrothermal carbonization of sewage sludge. METHODS AND RESULTS: The removal of human pathogenic rotavirus and adenovirus was investigated with columns of 10 cm saturated sand with and without amendments of 1·5% (w/w) hydrochar. Virus concentrations were determined with reverse transcription (RT) quantitative polymerase chain reaction (qPCR). The experiments with sand showed 1 log removal, while the columns with 1·5% (w/w) hydrochar amendment showed 2 to >3 log removal for both viruses. Deionized (DI) water flushing into the virus-retaining columns revealed that the secondary energy minimum played a larger role in the attachment of rotavirus onto hydrochar surfaces than adenovirus. Improved virus removal may be attributed to the introduction of hydrophobic and/or meso-macro surface structures of the hydrochar providing favourable attachment sites for viruses. CONCLUSIONS: Hydrochar amended sand beds showed improved virus removal efficiencies exceeding 99·6% corresponding to 2·4 log removal. The addition of humic acid in the influent did not hinder the adsorptive removal of viruses. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that hydrochar derived from sewage sludge can be used as an adsorbent for virus removal in water treatment.

Donor Heart Treatment With COMP-Ang1 Limits Ischemia-Reperfusion Injury and Rejection of Cardiac Allografts.

The major cause of death during the first year after heart transplantation is primary graft dysfunction due to preservation and ischemia-reperfusion injury (IRI). Angiopoietin-1 is a Tie2 receptor-binding paracrine growth factor with anti-inflammatory properties and indispensable roles in vascular development and stability. We used a stable variant of angiopoietin-1 (COMP-Ang1) to test whether ex vivo intracoronary treatment with a single dose of COMP-Ang1 in donor Dark Agouti rat heart subjected to 4-h cold ischemia would prevent microvascular dysfunction and inflammatory responses in the fully allogeneic recipient Wistar Furth rat. COMP-Ang1 reduced endothelial cell-cell junction disruption of the donor heart in transmission electron microscopy during 4-h cold ischemia, improved myocardial reflow, and reduced microvascular leakage and cardiomyocyte injury of transplanted allografts during IRI. Concurrently, the treatment reduced expression of danger signals, dendritic cell maturation markers, endothelial cell adhesion molecule VCAM-1 and RhoA/Rho-associated protein kinase activation and the influx of macrophages and neutrophils. Furthermore, COMP-Ang1 treatment provided sustained anti-inflammatory effects during acute rejection and prevented the development of cardiac fibrosis and allograft vasculopathy. These results suggest donor heart treatment with COMP-Ang1 having important clinical implications in the prevention of primary and subsequent long-term injury and dysfunction in cardiac allografts.

Angiopoietin-2 inhibition prevents transplant ischemia-reperfusion injury and chronic rejection in rat cardiac allografts.

Transplant ischemia-reperfusion injury (Tx-IRI) and allograft dysfunction remain as two of the major clinical challenges after heart transplantation. We investigated the role of angiopoietin-2 (Ang2) in Tx-IRI and rejection using fully MHC-mismatched rat cardiac allografts. We report that plasma levels of Ang2 were significantly enhanced in the human and rat recipients of cardiac allografts, but not in the rat recipients of syngrafts, during IRI. Ex vivo intracoronary treatment of rat cardiac allografts with anti-Ang2 antibody before 4-h cold preservation prevented microvascular dysfunction, endothelial cell (EC) adhesion molecule expression and leukocyte infiltration, myocardial injury and the development of cardiac fibrosis and allograft vasculopathy. Recipient preoperative and postoperative treatment with anti-Ang2 antibody produced otherwise similar effects without effecting microvascular dysfunction, and in additional experiments prolonged allograft survival. Recipient preoperative treatment alone failed to produce these effects. Moreover, ex vivo intracoronary treatment of allografts with recombinant Ang2 enhanced Tx-IRI and, in an add-back experiment, abolished the beneficial effect of the antibody. We demonstrate that neutralization of Ang2 prevents EC activation, leukocyte infiltration, Tx-IRI and the development of chronic rejection in rat cardiac allografts. Our results suggest that blocking Ang2 pathway is a novel, clinically feasible, T cell-independent strategy to protect cardiac allografts.

Impact of radical and partial nephrectomy on renal function in patients with renal cancer.

OBJECTIVE: To evaluate renal function in renal cancer patients undergoing radical nephrectomy (RN) or partial nephrectomy (PN) (open or laparoscopic - ORN, OPN, LRN or LPN) and to identify risk factors contributing to renal function loss. METHODS: We analysed 228 consecutive renal cancer patients admitted for OPN, LPN, ORN or LRN. The variables analysed were age, gender, weight, type of surgery (radical versus partial), type of surgical access (open versus laparoscopic), preoperative renal function and history of hypertension, diabetes or malignancy. Absolute renal function was calculated as the difference in glomerular filtration rate (ΔGFR) between the renal function before (GFR0) and 12 months after surgery (GFR12). The relative renal function of patients undergoing PN and RN was evaluated by the change in chronic kidney disease stage. RESULTS: LRN caused the greatest loss in absolute renal function, followed by ORN, LPN and OPN. A GFR of ≥60 ml/min was noted for 90 (68.7%) patients before and 65 (49.6%) patients after RN and for 80 (82.5%) patients before and 74 (76.3%) patients after PN. The chronic kidney disease stage dropped to 4 or 5 in the case of 6 (4.6%) patients who underwent RN and 2 (2.1%) patients who underwent PN. Multivariate analysis revealed that only preoperative weight and type of surgery (radical versus partial) had a significant impact on renal function. CONCLUSION: Renal function significantly decreased in patients undergoing RN, irrespective of the access route. Patients with preoperative poor renal function are at risk of postoperative end-stage renal disease.

Donor simvastatin treatment prevents ischemia-reperfusion and acute kidney injury by preserving microvascular barrier function.

Ischemia-reperfusion injury (IRI) after kidney transplantation may result in delayed graft function. We used rat renal artery clamping and transplantation models to investigate cholesterol-independent effects of clinically relevant single-dose peroral simvastatin treatment 2 h before renal ischemia on microvascular injury. The expression of HMG-CoA reductase was abundant in glomerular and peritubular microvasculature of normal kidneys. In renal artery clamping model with 30-min warm ischemia, simvastatin treatment prevented peritubular microvascular permeability and perfusion disturbances, glomerular barrier disruption, tubular dysfunction and acute kidney injury. In fully MHC-mismatched kidney allografts with 16-h cold and 1-h warm ischemia, donor simvastatin treatment increased the expression of flow-regulated transcription factor KLF2 and vasculoprotective eNOS and HO-1, and preserved glomerular and peritubular capillary barrier integrity during preservation. In vitro EC Weibel-Palade body exocytosis assays showed that simvastatin inhibited ischemia-induced release of vasoactive angiopoietin-2 and endothelin-1. After reperfusion, donor simvastatin treatment prevented microvascular permeability, danger-associated ligand hyaluronan induction, tubulointerstitial injury marker Kim-1 immunoreactivity and serum creatinine and NGAL levels, and activation of innate and adaptive immune responses. In conclusion, donor simvastatin treatment prevented renal microvascular dysfunction and IRI with beneficial effects on adaptive immune and early fibroproliferative responses. Further studies may determine potential benefits in clinical cadaveric kidney transplantation.

Evaluation of the Joypex 5 and Root ZX II: an in vivo and ex vivo study.

AIM: To evaluate in vivo and ex vivo the accuracy of the multifrequency electronic root canal length measurement device (ERCLMD) Joypex 5 and compare it with the Root ZX II. The ex vivo methodology for evaluation of the accuracy of ERCLMDs was also validated by comparison with in vivo results. METHODOLOGY: Twenty single-rooted human teeth that were scheduled for extraction were selected. Working length measurements were performed in vivo with the Joypex 5 and Root ZX II. After the teeth were extracted, each canal was measured with the ERCLMDs ex vivo. Then, a size 15 K-file was used to determine the reference working length (RWL), which was established 0.5 mm from the major foramen by direct observation. The measurements of working length obtained with the different methods and ERCLMDs were compared by analysis of variance and Tukey's test at P < .05. Statistical correlations were also performed. RESULTS: No significant difference was observed between in vivo and ex vivo measurements or the different types of ERCLMDs. The Bland-Altman plot confirmed the agreement of different methods and ERCLMDs to measure canal length. The R(2) coefficient obtained in both situations, comparing the ERCLMDs, was close to 1, denoting a strong agreement between measurements obtained with the Joypex 5 and Root ZX II and between in vivo and ex vivo methodologies. CONCLUSION: The Joypex 5 and Root ZX II had similar accuracy in determining working length of root canals. The ex vivo methodology for evaluation of the accuracy of ERCLMDs was validated when compared with the in vivo results.

Differential effects of pharmacological HIF preconditioning of donors versus recipients in rat cardiac allografts.

Ischemia-reperfusion injury (IRI) induces hypoxia-inducible factor-1 (HIF-1) in the myocardium, but the consequences remain elusive. We investigated HIF-1 activation during cold and warm ischemia and IRI in rat hearts and cardiac syngrafts. We also tested the effect of HIF-α stabilizing prolyl hydroxylase inhibitor (FG-4497) on IRI or allograft survival. Ex vivo ischemia of the heart increased HIF-1α expression in a time- and temperature-dependent fashion. Immunohistochemistry localized HIF-1α to all cardiac cell types. After reperfusion, HIF-1α immunoreactivity persisted in smooth muscle cells and cardiomyocytes in the areas with IRI. This was accompanied with a transient induction of protective HIF-1 downstream genes. Donor FG-4497 pretreatment for 4 h enhanced IRI in cardiac allografts as evidenced by an increase in cardiac troponin T release, cardiomyocyte apoptosis, and activation of innate immunity. Recipient FG-4497 pretreatment for 4 h decreased infiltration of ED1(+) macrophages, and mildly improved the long-term allograft survival. In syngrafts donor FG-4497 pretreatment increased activation of innate immunity, but did not induce myocardial damage. We conclude that the HIF-1 pathway is activated in heart transplants. We suggest that pharmacological HIF-α preconditioning of cardiac allografts donors would not lead to clinical benefit, while in recipients it may result in antiinflammatory effects and prolonged allograft survival.

Novelty increases the mesolimbic functional connectivity of the substantia nigra/ventral tegmental area (SN/VTA) during reward anticipation: Evidence from high-resolution fMRI.

Reward and novelty are potent learning signals that critically rely on dopaminergic midbrain responses. Recent findings suggest that although reward and novelty are likely to interact, both functions may be subserved by distinct neuronal clusters. We used high-resolution functional magnetic resonance imaging (fMRI) to isolate neural responses to reward and novelty within the human substantia nigra/ventral tegmental area (SN/VTA) complex to investigate the spatial delineation and integration of reward- and novelty-related activity clusters. We demonstrate that distinct clusters within the caudal portion of the medial SN/VTA and the lateral portion of the right SN are predominantly modulated by the anticipation of reward, while a more rostral part of the medial SN/VTA was exclusively modulated by novelty. In addition, the caudal medial SN/VTA cluster embodied an interaction between novelty and reward where novelty selectively increased reward-anticipation responses. This interaction, in turn, was paralleled by differences in the functional-connectivity patterns of these SN/VTA regions. Specifically, novel as compared to familiar reward-predictive stimuli increased the functional connectivity of the medial SN/VTA with mesolimbic regions, including the nucleus accumbens and the hippocampus, as well as with the primary visual cortex. This functional correlation may highlight how afferents of the medial SN/VTA provide integrative information about novelty and reward, or, alternatively, how medial SN/VTA activity may modulate memory processes for novel events associated with rewards.

Pinning down response inhibition in the brain--conjunction analyses of the Stop-signal task.

Successful behavior requires a finely-tuned interplay of initiating and inhibiting motor programs to react effectively to constantly changing environmental demands. One particularly useful paradigm for investigating inhibitory motor control is the Stop-signal task, where already-initiated responses to Go-stimuli are to be inhibited upon the rapid subsequent presentation of a Stop-stimulus (yielding successful and unsuccessful Stop-trials). Despite the extensive use of this paradigm in functional neuroimaging, there is no consensus on which functional comparison to use to characterize response-inhibition-related brain activity. Here, we utilize conjunction analyses of successful and unsuccessful Stop-trials that are each contrasted against a reference condition. This conjunction approach identifies processes common to both Stop-trial types while excluding processes specific to either, thereby capitalizing on the presence of some response-inhibition-related activity in both conditions. Using this approach on fMRI data from human subjects, we identify a network of brain structures that was linked to both types of Stop-trials, including lateral-inferior frontal and medial frontal cortical areas and the caudate nucleus. In addition, comparisons with a reference condition matched for visual stimulation identified additional activity in the right inferior parietal cortex that may play a role in enhancing the processing of the Stop-stimuli. Finally, differences in stopping efficacy across subjects were associated with variations in activity in the left anterior insula. However, this region was also associated with general task accuracy (which furthermore correlated directly with stopping efficacy), suggesting that it might actually reflect a more general mechanism of performance control that supports response inhibition in a relatively nonspecific way.

Sensory MEG responses predict successful and failed inhibition in a stop-signal task.

In the present study magnetoencephalographic recordings were performed to investigate the neural mechanisms underlying the stopping of manual responses. Subjects performed in a Stop-signal task in which Go-stimuli (S1), requiring a rapid motor response, were sometimes rapidly followed by a Stop-stimulus (S2) indicating to withhold the already initiated response to S1. Success of stopping strongly depended on the early perceptual processing of S1 and S2 reflected by the magnetic N1 component. Enhanced processing of S1 facilitated the execution of the movement, whereas enhanced processing of S2 favored its inhibition. This suggests that the processing resources for the subsequent stimuli are limited and need to be shared. This sharing of resources appeared to arise from adjustments made on a trial-by-trial basis, in that systematic reaction time prolongations on Go-trials following Stop-trials versus following Go-trials were accompanied by attenuated sensory processing to the Go-stimulus similar to that seen in successful versus unsuccessful stopping in Stop-trials.

Vascular endothelial growth factor plays a major role in development of experimental obliterative bronchiolitis.

Obliterative bronchiolitis (OB) is the major limitation for long-term survival of lung allograft recipients. The exact molecular and cellular mechanisms contributing to obliterative lesion formation are unknown. Pathological characteristics of OB are epithelial damage, peribronchial inflammation, and increasing obliteration of bronchioli. Vascular endothelial growth factor (VEGF) is an angiogenic growth factor that exerts proinflammatory effects by increasing endothelial permeability and inducing expression of endothelial adhesion molecules. We investigated the role of VEGF in the development of OB in rat tracheal allografts and the role of VEGF receptors (VEGFR)-1 and -2 in the development of OB in mouse tracheal allografts. In nontreated allografts, with increasing loss of epithelium and airway occlusion, VEGF messenger RNA (mRNA) and protein expression vanished in the epithelium and increased in smooth muscle cells and mononuclear inflammatory cells compared with syngeneic grafts. Intragraft VEGF overexpression by adenoviral transfer of a mouse VEGF164 gene led to a decrease in epithelial necrosis but increased luminal occlusion by >50% compared with AdLacZ-treated rat tracheal allografts. When compared with the control immunoglobulin (Ig)G group, simultaneous treatment with antibodies against VEGFR-1 and -2 significantly lowered the degree of luminal occlusion of mouse tracheal allografts.

The effect of platelet-derived growth factor ligands in rat cardiac allograft vasculopathy and fibrosis.

BACKGROUND: In chronic rejection, parenchymal fibrosis and cardiac allograft vasculopathy (CAV) characterized by neointimal growth are the leading causes of graft loss for heart transplant recipients. During alloimmune responses a variety of cytokines, adhesion proteins, and growth factors, such as platelet-derived growth factor (PDGF), are up-regulated. The PDGF family (AA, AB, BB, CC, DD), which acts mainly on connective tissue cells, is considered to be a potent mitogenic and chemotactic factor for fibroblasts and vascular smooth muscle cells. In this study, we evaluated the effects of PDGF ligands in chronic rejection. METHODS: Heterotopic heart transplantations were performed between fully major histocompatability complex-mismatched Dark Agouti to Wistar Furth rats receiving cyclosporine immunosuppression. Allograft coronary arteries were perfused with a recombinant adeno-associated virus (AAV) encoding enhanced green fluorescence protein (EGFP) as a control gene or PDGF-A, -B, -C, -D. Allografts were harvested at 100 days for morphometric analysis of CAV and fibrosis. RESULTS: AAV-mediated transgene expression was detected by EGFP immunoreactivity across the graft section (at 100 days) in AAV EGFP-perfused allografts. AAV PDGF-A, -C, and -D perfusion resulted in accelerated CAV and fibrosis. In contrast, AAV PDGF-B perfusion did not induce arteriosclerotic changes or fibrosis in cardiac allografts. CONCLUSIONS: AAV PDGF-A, -C, and -D overexpression accelerated the development of chronic rejection, whereas PDGF-B did not. Our results suggested that more targeted therapy with monoclonal antibodies blocking the active sites of PDGF-A, -C, and -D may produce beneficial effects on heart transplant survival.

Effect of graft preservation and acute rejection on hypoxia-inducible factor-1 in rat cardiac allografts.

Hypoxia plays an integral part in cardiac transplantation as prolonged graft preservation is an individual risk factor for the development of cardiac allograft vasculopathy (CAV). In this study we characterized the role of hypoxia-inducible factor-1 (HIF-1) during prolonged graft preservation, ischemia-reperfusion (I/R), acute rejection, and chronic rejection. Heart transplantations were performed from Dark Agouti (DA) to Wister-Furth (allo) or DA to DA (syn) rats, without immunosuppression (acute rejection model, harvested at day 5) or with cyclosporine (chronic rejection model, harvested at day 60). To study the effect of preservation on HIF-1 regulation, normal DA hearts were subjected to different cold ischemia times with or without 45 minutes of additional warm ischemia. The role of I/R was studied by harvesting syngrafts at different time points after reperfusion. Real-time reverse-transcriptase polymerase chain reaction quantified total HIF-1 mRNA, while enzyme-linked immunosorbent assay and immunohistochemistry quantified and localized HIF-1 protein. Our results show that HIF-1 nuclear immunoreactivity is increased during graft preservation and I/R leads to loss of nuclear HIF-1 immunoreactivity. Acute rejection induced HIF-1 in mRNA level. Our findings thus indicated that HIF-1 is activated during transplantation and suggested that manipulation of the HIF-1 pathway might reveal new therapeutic options to manage CAV.

The effect of thermal stress on the mating behavior of three Drosophila species.

Selection may act on the weakest link in fitness to change how a species adapts to an environmental stress. For many species, this limitation may be reproduction. After adult Drosophila melanogaster, Drosophila simulans, and Drosophila mojavensis males were exposed to varying levels of thermal stress well below those that endanger life, courtship and mating frequency declined. The regression coefficients of both courtship and mating success did not differ significantly between D. melanogaster and D. simulans males. In contrast, significant differences were present between the two cosmopolitan species and D. mojavensis. Courtship frequency decreased at a much slower rate in D. mojavensis than in D. melanogaster and D. simulans, and while heat-stressed D. mojavensis males continued to court, many did not mate. In the cosmopolitan species, courting males almost always mated successfully. Courtship behaviors, including wing waving, were observed in D. mojavensis at temperatures that prohibited flight, while flight, courtship, and mating were knocked out simultaneously in D. melanogaster. One possible explanation for decreased flight ability and courtship success may be the reduced heat shock response in the flight muscle tissue because Hsp70 expression was lowest in the thoracic tissue of both D. melanogaster and D. mojavensis.

Reduced enzyme activity following Hsp70 overexpression in Drosophila melanogaster.

Acclimation to environmental change can impose costs to organisms. One potential cost is the change in cell metabolism that follows a physiological response, e.g., high expression of heat shock proteins may alter specific activity of important enzymes. We examined the significance of this cost in a pair of Drosophila melanogaster lines transformed with additional copies of a gene that encodes the heat shock protein, Hsp70. Heat shock induces Hsp70 expression in all lines, but lines with extra copies produce much more Hsp70 than do excision control strains. The consequence of this supranormal Hsp70 expression is to reduce specific activity of both enzymes analyzed, adult alcohol dehydrogenase (ADH), which is heat sensitive, and lactate dehydrogenase, which is not. Strain differences were most pronounced under those conditions where Hsp70 expression was maximized, and not where the heat stress denatured proteins. That result supported the idea that Hsp70 expression is constrained evolutionarily by its tendency to bind nascent peptides when overabundant within the cell.

Activin A and activin receptors in thyroid cancer.

Proliferation is controlled by a network of mitogenic and growth inhibitory factors. Transforming growth factor-beta1 (TGF-beta1) and activin A are the most important growth inhibitors of benign follicular epithelial cells of the human thyroid. The effects of these substances on malignant primary thyrocytes are not known. We have examined the growth regulatory effects of activin A and TGF-beta1 in primary cultures derived from four papillary cancers, two follicular thyroid cancers, and three benign thyroid tissues. Malignant cells demonstrated resistance to activin and TGF-beta1 or reversal to a weak but significant mitogenic effect (p < 0.001). We also evaluated the activin receptor transcription pattern. Isoforms alk4-1, 4-2, and 4-3 were found in benign (n = 12) and malignant (n = 22) tissues. Two subtypes of type I and type II activin receptors were demonstrated. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) demonstrated a significant threefold downregulation of alk4-1 receptors in papillary (n = 25) and follicular (n = 18) thyroid cancers as compared to normal thyroids (n = 12) (p < 0.001). To our knowledge these are the first data to demonstrate reversal of activin and TGF-beta1 effects in thyroid malignancy and to demonstrate changes of the type Ib activin receptor expression in thyroid malignancy.