In-vivo Changes Research Articles

Histological findings of thyroid cancer after lenvatinib therapy.

Lenvatinib is a multikinase inhibitor used for treating unresectable or metastatic cancers, including thyroid cancer. As total thyroidectomy followed by radioactive iodine therapy is a commonly recommended initial treatment for thyroid cancer, histological findings of the thyroid after lenvatinib therapy remain unclear. Therefore, the aim of this study was to analyse in-vivo changes in patients who underwent thyroidectomy after lenvatinib therapy. We screened 167 patients with thyroid cancer [papillary thyroid cancer (PTC), n = 102; follicular thyroid cancer (FTC), n = 26; anaplastic thyroid cancer (ATC), n = 39] who underwent lenvatinib therapy. Among these patients, six underwent thyroidectomy (lenvatinib-treated group: PTC, n = 3; FTC, n = 1; ATC, n = 2), and the specimens were examined. Five patients with PTC who did not receive lenvatinib therapy were included for comparison (untreated group). Microvessel density (MVD) was evaluated in both groups. The PTC and FTC specimens showed relatively more ischaemic changes than ATC specimens. Coagulative necrosis and ischaemic changes in cancer cells were frequently observed. ATC specimens showed fibrosis and mild cell damage. As hypothyroidism is a common side effect of lenvatinib therapy, non-cancerous thyroid tissues were also examined. Histological findings included mild lymphocytic infiltration, lymphoid follicular formation, histiocytic reaction and follicular epithelial destruction. The MVD in lenvatinib-treated tissues was significantly lower than that in untreated tissues. Lenvatinib therapy probably induces relatively specific ischaemic changes in thyroid cancer cells. Moreover, inflammatory cell infiltration and decreased MVD occur to varying degrees in non-cancerous thyroid tissue and may be related to hypothyroidism, a side effect of lenvatinib.

Preconcentration, Release and Electrochemical Detection of microRNA on Microfluidics

Introduction One of the challenges of the 21st in medical diagnosis lies in the identification of specific biomarkers for the detection of infectious or cancerous diseases before the first symptoms appear and thus treat the patients effectively.After the discovery of their implication in cancer pathologies [1] at early stage, micro ribonucleic acids (miRNA) became promising candidates for this application due to their rapid, precise and reliable diagnosis. These miRNAs, constituted by around 20 nucleotides, are highly involved in post-transcriptional regulation mechanisms capable of inducing the extinction or expression of certain gens and their presence in body fluids (plasma, urine, saliva...) is a sign of important perturbations [2].However, in order to detect the in-vivo change of molecular expression levels (under femtomolar [3]) current techniques rely in chemical amplification stages of a specific target to be detected usually done by Polymerase Chain Reaction (PCR). The main drawbacks of PCR are an increase in the entire protocol time from miRNA extraction in biological sample to its detection (optical or electrical), as well as nonspecific sequence amplification, especially for short sequences as in miRNAs [4].The approach developed and patented by our laboratory [5] proposes an alternative to the use of chemical amplification techniques. This alternative is based in the preconcentration of the circulating miRNA, their localized release and detection. During the preconcentration stage, functionalized magnetic nanoparticles (NPMs) capture the circulating miRNAs and then release them, via magnetic hyperthermia. Once released, the miRNAs are detected by functionalized gold microelectrodes embedded in a microfluidic chip [6].In this communication we will present the results obtained combining the localized release of the miRNA and their electrochemical detection. Method The used nanoparticles have a diameter of 80 nm and they are formed by a maghemite core protected by a silica shell functionalized with –NH2 groups which allow a covalent coupling with modified RNA probe.The microelectrode pairs used to detect the hybridation of the previously released miRNA are formed by a 30 µm width by 300 µm length working gold microelectrode and a 30 µm width by 2 mm length gold counter-electrode patterned on a glass substrate by a lift-off process.The microfluidic channels are microfabricated by molding dimethylpolysiloxane (PDMS) onto a SU-8 (from MicroChem) mold 50-micron thickness with the pattern of the channels. PDMS is spin coated on the mold to obtain a thickness of 500 µm which allows to keep the total thickness of the microfluidic device under 1mm after the bonding of the glass substrate and the PDMS microchannels.Functionalization of the microelectrodes is done by introducing a solution of DNA probes modified with a thiol termination in 0.5 M NaCl. Then the channel is rinsed with 0.5 M NaCl to stabilize the probes.The magnetic field needed for the magnetic hyperthermia is generated by a power amplifier that provides the current to the tank circuit formed by a capacitor and an air-gapped ferrite with a certain number of turns. The microfluidic device is placed in the air-gap of the ferrite to maximize the amplitude of the applied alternating magnetic fieldElectrochemical measurements are performed using a commercial potentiostat connected to the microfluidic device through a dedicated chip holder. Electrochemical measurements are performed in 0.5 M NaCl as supporting electrolyte containing a 3 mM FeIIICN6 3-/ FeIICN6 4- redox couple, and 10-8 M methylene blue as redox intercalant. Results and Conclusions The measured impedance between a working microelectrode and the counter-electrode before and after the hybridation of the miRNA released by the magnetic nanoparticles after being subjected to an alternating magnetic field as previously described [5].In this presentation we will present impedance measurements before hybridation that shows a high electron charge transfer resistance (Rct) since grafted DNA probe (single strands) inhibits the redox complex diffusion towards the electrodes. After the miRNA hybridization on microelectrodes (double strand), the electron charge transfer resistance decreases related to the intercalation of MB in DNA duplex which acts as an electron charge mediator between the redox and the microelectrode. Indeed, Rct decrease highlights the success of the hybridization step after it release from magnetic hyperthermia runs.The proposed methodology, based in the combination of magnetic hyperthermia and electrochemical detection, has shown to be effective to locally release the recruited miRNA and detect them by electrochemical means. In addition, the whole process is performed in a microfluidic device offering the advantages of this devices: batch processing, low sample volume, portability, etc.

In-vivo change of the spine canal after surgical corrections of severe and rigid kyphoscoliosis

Objective: To measure the length changes of the spine canal of patients with severe kyphosis after treatments of deformity using osteotomy surgeries. Methods: Retrospectively investigated the data of 10 severe kyphosis patients who were treated between August of 2016 and December of 2018 at Beijing Chaoyang Hospital (5 with Ponte and 5 with posterior vertebra column resection (VCR) osteotomy surgeries). For each patient, the full-spine X-Ray images were used to measure Cobb angles before and after the surgery; 3D CT images were used to construct a 3D model of the spine, including the T(2) to L(2) vertebrae and the spine canal. The 3D model was then used to measure the spinal canal lengths (SCLs) between the upper and lower vertebrae (U/LEV) and between T(2)-L(2) vertebrae at 5 locations on the spine canal cross section (anterior, central, posterior, left and right locations), and the vertical distance between the T(2) and L(2) vertebrae before and after the surgery. The data were statistically analyzed using t tests. Results: For the 5 patients of Ponte osteotomy, the Cobb angles were improved by 89°±17°(56%±11%) and 84°±16°(56%±8%) in the coronal and sagittal planes respectively after the surgery. The changes of the SCL between the T(2)-L(2) vertebrae were (9.9±4.8) mm and (6.0±12.7) mm, and those were (7.2±5.4) mm and (-0.5±7.9) mm between the U/LEV, respectively at the concave and convex sides of the canal. The vertical distance between the T(2)-L(2) vertebrae increased by (66.1±12.0) mm. For the 5 patients with VCR osteotomy, the Cobb angles improved by 83°±19°(60%±10%) and 82°±22°(56%±10%) in the coronal and sagittal planes, respectively. The changes of the SCL between the T(2)-L(2) vertebrae were (-5.5±5.3) mm and (-14.0±6.6) mm, and those were (-8.3±8.4) mm and (-20.7±11.6) mm between the U/LEV, respectively at the concave and the convex sides of the canal. The vertical distance between the T(2)-L(2) vertebrae increased by (41.5±12.4) mm. Conclusions: The Ponte osteotomy significantly elongates the SCLs, especially at the concave side, and the VCR osteotomy shortens the spinal canal, resulting in significant compression of the spinal cord at the convex side.

Clinical outcome and ultrasonographic evaluation of treatment using polyglycolic acid-collagen tube for chronic neuropathic pain after peripheral nerve injury

BackgroundA novel polyglycolic acid (PGA)-collagen tube has been available in Japan since March 2013, and the development of high-resolution ultrasonography has recently allowed detailed evaluation of nerves. The purpose of this study was to report the clinical outcomes and evaluations of in-vivo changes under high-resolution ultrasonography for treatment using PGA-collagen tube for painful chronic nerve injury. MethodsFour patients underwent surgical management of peripheral nerve neuroma, with nerve defects after neuroma resection reconstructed using PGA-collagen tube. Mean duration of follow-up was 30 months. Patients were surveyed to determine visual analog scale scores for pain before surgery and at final follow-up. At final follow-up, sensory function was evaluated using the Semmes–Weinstein test, static 2-point discrimination (2PD), and moving 2PD. Furthermore, we performed ultrasonography for all patients every 1 month after surgery. ResultsIn all cases, resting pain, radicular pain, and dysesthesia were improved immediately after the operation and remained improved until the last follow-up. At the time of final follow-up, the Semmes–Weinstein test was red in 2 cases and blue in 2 cases. Mean static 2PD was 15.5 ± 5.3 mm and mean moving 2PD was 11.5 ± 5.3 mm. Ultrasonography revealed that the PGA outer wall had disappeared by 3 months postoperatively, and a hypoechoic linear shadow bridged the proximal and distal stumps. Swelling of the junction between nerve and nerve conduits was revealed at 4 months after surgery and remained until last follow-up. ConclusionsAll patients were very satisfied with pain relief, but recovery of sensory function was insufficient. Ultrasonographic evaluation after using PGA-collagen tube for painful chronic peripheral nerve injury was useful for observing in-vivo changes. Type of study/level of evidenceTherapeutic IV.

In Vivo Effects of Long-Term Cigarette Smoke Exposure on Mammary Tissue in Mice

Cigarette smoking is the leading cause of preventable death worldwide and has been linked to the development and progression of cancer. Many cohort studies have described the link between patients with breast cancer and those with long-term smoking history. Despite the claim of correlation, the mechanism by which cigarette smoke alters normal breast epithelial cells and stroma and contributes to tumor cell growth remains undefined. To investigate whether cigarette smoke promotes ductal epithelial cell hyperplasia by stimulating stromal endothelial cell proliferation, we exposed mice to cigarette smoke for 6 months. We observed epithelial proliferation, increased fibrosis, increased vascularity, and mast cell infiltration. This is the first study to look at the invivo changes in the breast after long-term cigarette smoke exposure and provides a novel insight to understanding how cigarette smoke contributes to early changes that may contribute to tumor formation and progression. In conclusion, this study suggests that cigarette smoke modulates key stromal-epithelial interactions to support increased angiogenesis, desmoplasia, and abnormal ductal epithelial cell growth.

Loperamide-induced Cardiac Depression Is Enhanced by Hyperglycemia: Evidence Relevant to Loperamide Abuse

Cardiac dysryhthmias and death are reported after loperamide abuse. The mechanism of death is not clear and cardiac depression may play a role in this mechanism. Loperamide is widely used as an agonist of the μ-opioid receptor (MOR) in clinical practice. In skeletal muscle, an increase in MOR in response to hyperglycemia is largely attributable to higher expression of the transducer and activator of transcription 3 (STAT3), which binds to the promoter of the MOR genes. Therefore, we investigated the changes in cardiac MOR caused by hyperglycemia both invivo and invitro. Streptozotocin-induced type 1-like diabetic rats (STZ rats) were used to estimate cardiac performance and changes in cardiac MOR under the influence of loperamide. STAT3 was measured in cultured cardiomyocytes under high glucose (HG) to mimic the invivo changes. Loperamide-induced reduction of cardiac performance was more marked in STZ rats than in normal rats. The increased MOR in the hearts of STZ rats was reversed by the reduction of hyperglycemia. Higher MOR expression paralleled the increase in STAT3 in cardiomyocytes under HG and was reversed by siRNA of STAT3. Stattic at a dose sufficient to inhibit STAT3 reduced MOR both invivo and invitro. Cardiac depression induced by loperamide is enhanced by hyperglycemia due to higher MOR expression, which is associated with higher expression of STAT3 in the heart. These results suggest that loperamide abuse is particularly dangerous for individuals with hyperglycemia.

Optical monitoring of retinal respiration in real time: 670 nm light increases the redox state of mitochondria

Mitochondria play a key role in ageing and disease. Their membrane potentials and ATP production decline with age and this is associated with progressive inflammation, cell loss and death. Here we use broadband Near-Infrared Spectroscopy (NIRS) to non-invasively measure in-vivo changes in aged retinal mitochondrial respiration following exposure to 670 nm, which improves mitochondrial performance and reduces inflammation. Low power NIR light was shone into the eye via a fibre optic and the reflection monitored to measure signature changes in the oxidation of cytochrome c oxidase (COX) in complex IV of the electron transport chain. Changes in retinal haemodynamics and oxygenation were also recorded simultaneously with COX by measuring changes in oxygenated and deoxygenated haemoglobin (Δ[HbO2] and Δ[HHb]). Retinae of aged rats exposed to 670 nm for 5 mins showed consistent progressive increases in oxidation of COX 5 mins post exposure. This remained significantly greater than baseline for up to 2 h. This was not seen when retinae were exposed to 420 nm light of the same power or when no light was applied. 670 nm exposure significantly increased total haemoglobin concentration (Δ[HbT] = Δ[HbO2] +Δ[HHb]) but not haemoglobin difference (Δ[HbDiff] = Δ[HbO2] -Δ[HHb]). There were no changes in blood metrics in association with 420 nm light or when no light exposure was given. Hence, brief 670 nm exposure that is associated with reduced inflammation has a significant positive impact on the redox state of COX in aged retinae. The relative redox state of retinal COX may provide a valuable biomarker in ageing and macular degeneration where declining mitochondrial function is implicated.

Http://www.omicsgroup.org/journals/polyolefin-from-commodity-to-specialty-2169-0022-1000e111.php?aid=47818

Generally, the combination use of difference metals occurs galvanic corrosion. Therefore, the combination use of SUS316L (SUS) and Ti-6Al-4V(Ti64) is supposed to be prohibited in human body. However, there are necessary cases of the combination use in clinical orthopedics. In order to investigate the in-vivo galvanic corrosion, the plates and screws made of SUS and Ti64 were implanted into the proximal tibia of the rabbits for 52 weeks. The plate and screws were implanted in 4 different combinations of pairing up in different metals and identical metals. By means of X-ray follow-up and Contact Micro Radiography (CMR), new bone formation around the implants was investigated. And cytopathological observation was performed on the surrounding tissue. Regarding to the implants, microscopic observation was performed on the removed implants surface. Furthermore, metal element analysis was carried out to investigate chromium ion release as a sign of the galvanic corrosion. In X-ray follow-up and CMR, the bone tissue under the plates became thin and porous like as concellous bone in all the combinations, which is supposedly due to the stress shielding and the disturbance of periosteal blood flow by the implantation. In the surrounding soft tissue and bone tissue, no remarkable findings were obtained in the cytopathological observation. Regarding to microscopic observation of the implants, short striations with metallic luster in SUS implants, and short striations with metallic luster or dark brown color in Ti64 implant were confirmed. However these are supposedly caused by the scratching or frictional damage during the screw fixation. In metal element analysis, the Cr element elution was confirmed around the SUS plates. However, there is no significant difference between SUS screw (identical metals) and Ti64 screw (different metals). As the results, the in-vivo galvanic corrosion and related changes were not confirmed in all investigations at 52 weeks.

Changes in the T and B- Cell Subsets Following Treatment with a CD25-Antibody in Patients with Steroid Refractory GvHD

Allogeneic stem cell transplantation is the therapeutic option for a variety of malignant and non-malignant haematological diseases. Graft versus Host Disease (GvHD) is a common post transplant complication. In 40% of these patients, GvHD is steroid refractory and associated with a mortality of around 60%. Basiliximab is a chimeric murine –human antibody also selective for interleukin -2 receptor (IL-2R) with a half life of 7 days. It is routinely used as part of the induction therapy in renal transplant recipients to prevent acute rejection following successful phase III studies. Phase 2 studies have demonstrated its superior efficacy in treating patients with steroid refractory GvHD (1). We administered Basiliximab in 14 patients with steroid refractory GvHD with a median age of 41 (range 20-69). M: F 7:7. All patients but one 13/14 received PBSC from unrelated donors and 6/13 had mismatched unrelated donors. Overall response was in the order of 12/14 (85%). One patient could not be assessed. 7/14 (50%) achieved a complete response to treatment. We aimed to study the in vivo T- and B-cell changes following Basiliximab administration as this would be an ideal platform to monitor the alterations in the regulatory T and B-cell compartment.PBMCs were obtained from all donors after informed consent, Immucan (Nr 11-116) approved by our local ethics committee, prior to and after weekly administration of Basiliximab 40mg for 4 weeks. Control samples were obtained from patients with steroid responsive acute GvHD.The total number of CD3+ as well as CD4+ and CD8+ T-cells remained constant during treatment and no change was observed on comparison with the controls. Gagliani et al (2) demonstrated that regulatory type 1 T-cells can be identified by the co-expression of CD49b and Lag3. No difference was observed between the % CD49d+, Lag3+ T-cells in the control cohort and the treatment cohort prior to therapy, ie day 0. The % CD49d+, Lag3+ T-cells decreased during the treatment period (statistically significant) in comparison to the control cohort. Despite the use of the CD25-antibody, a small population of CD25+, CD127+ cells could be detected and this population correlated to the % CD49d+, Lag3+ T-cells. [Display omitted] [Display omitted] The % CD19+, CD20+ B-cells were similar prior to treatment in the treatment group and control. Following the first administration a rise was observed followed by a decline over the next 3 weeks. No changes were seen in the activated (CD20+, CD86+) and anergic B-cell subsets (CD20+, CD21-) during the observation period. The % of CD24high, CD27+ regulatory B-cells were found to be twice that seen in the controls. With treatment a decrease was seen in this population. The CD24high, CD38high transitional B-cells were also found to be higher than that seen in the controls. No change was observed in this subset with treatment. [Display omitted] This is the first attempt to study the in-vivo changes induced by a CD25 antibody in patients with steroid refractory GvHD. We conclude that this antibody not only depletes the alloreactive CD25+ T and B-cell population but also alters the regulatory T and B-cell subsets in comparison to patients with steroid responsive GvHD. Our clinical data supports the efficacy of this drug in patients with steroid refractory GvHD. Contrary to the current knowledge that regulatory T-cells are required for GvHD suppression our data suggests that Basiliximab facilitates regulatory T-cell depletion. The reduction of the regulatory T-cell subset observed in patients responding to anti CD25 treatment suggests a complex regulation and potential dichotomous role of these cells in acute GvHD. 1.Schmidt-Hieber M, Fietz T, Knauf W, Uharek L, Hopfenmuller W, Thiel E, et al. Efficacy of the interleukin-2 receptor antagonist basiliximab in steroid-refractory acute graft-versus-host disease. Br J Haematol. 2005 Aug;130(4):568-74.2.Gagliani N, Magnani CF, Huber S, Gianolini ME, Pala M, Licona-Limon P, et al.Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells. Nat Med. 2013 Jun;19(6):739-46 DisclosuresNo relevant conflicts of interest to declare.

Oncogenic Kras Activates a Hematopoietic-to-Epithelial IL-17 Signaling Axis in Preinvasive Pancreatic Neoplasia

Many human cancers are dramatically accelerated by chronic inflammation. However, the specific cellular and molecular elements mediating this effect remain largely unknown. Using a murine model of pancreatic intraepithelial neoplasia (PanIN), we found that Kras(G12D) induces expression of functional IL-17 receptors on PanIN epithelial cells and also stimulates infiltration of the pancreatic stroma by IL-17-producing immune cells. Both effects are augmented by associated chronic pancreatitis, resulting in functional invivo changes in PanIN epithelial gene expression. Forced IL-17 overexpression dramatically accelerates PanIN initiation and progression, while inhibition of IL-17 signaling using genetic or pharmacologic techniques effectively prevents PanIN formation. Together, these studies suggest that a hematopoietic-to-epithelial IL-17 signaling axis is a potent and requisite driver of PanIN formation.

Rapid and Pervasive Changes in Genome-wide Enhancer Usage during Mammalian Development

Enhancers are distal regulatory elements that canactivate tissue-specific gene expression and areabundant throughout mammalian genomes. Although substantial progress has been made toward genome-wide annotation of mammalian enhancers, their temporal activity patterns and global contributions in the context of developmental invivo processes remain poorly explored. Here we used epigenomic profiling for H3K27ac, a mark of active enhancers, coupled to transgenic mouse assays to examine the genome-wide utilization of enhancers in three different mouse tissues across seven developmental stages. The majority of the ∼90,000 enhancers identified exhibited tightly temporally restricted predicted activity windows and were associated with stage-specific biological functions and regulatory pathways in individual tissues. Comparative genomic analysis revealed that evolutionary conservation of enhancers decreases following midgestation across all tissues examined. The dynamic enhancer activities uncovered in this study illuminate rapid and pervasive temporal invivo changes in enhancer usage that underlie processes central to development and disease.

Analysis of In Vitro Insulin-Resistance Models and Their Physiological Relevance to In Vivo Diet-Induced Adipose Insulin Resistance

Diet-induced obesity (DIO) predisposes individuals to insulin resistance, and adipose tissue has a major role in the disease. Insulin resistance can be induced in cultured adipocytes by a variety of treatments, but what aspects of the invivo responses are captured by these models remains unknown. We use global RNA sequencing to investigate changes induced by TNF-α, hypoxia, dexamethasone, high insulin, and a combination of TNF-α and hypoxia, comparing the results to the changes in white adipose tissue from DIO mice. We found that different invitro models capture distinct features of DIO adipose insulin resistance, and a combined treatment of TNF-α and hypoxia is most able to mimic the invivo changes. Using genome-wide DNase I hypersensitivity followed by sequencing, we further examined the transcriptional regulation of TNF-α-induced insulin resistance, and we found that C/EPBβ is a potential key regulator of adipose insulin resistance.

123 In vivo evidence of COMT val158met functional polymorphism modulates different levels of dopamine turnover in Parkinson's disease: an 18F-dopa PET study

ObjectiveTo investigate in-vivo changes in dopamine turnover in Parkinson's disease (PD) with COMT val158met functional polymorphism using 18F-dopa positron emission tomography (PET).MethodsTwenty patients with PD (10 with val/val, and 10...

Prognosis of Low- or Intermediate-1-Risk MDS with Del(5q) Chromosome Abnormality During Lenalidomide Therapy Depends on the Capacity of the Neoplastic Stem Cell to Differentiate Into Maturing Erythroid Committed Stem Cells,

Abstract 3819Transfusion-dependent anemia is a characteristic complication of myelodysplastic syndromes (MDS) with del(5q) chromosome abnormality. Background appears to be a haplo-insufficiency of the RPS-14 gene located within the CDR. Lenalidomide has improved the treatment of MDS with 5q deletion inducing a hematologic (erythroid) and cytogenetic response in the majority of patients. The exact mechanisms of action of lenalidomide explaining its effects on erythropoiesis and their possible prognostic relevance are not clear so far. PATIENTS:We present the first data on in-vivo changes at the level of hematopoietic stem cells (SC), erythroid committed SC (SC-e), maturation of SC-e to erythroblastic islands and normoblasts observed during lenalidomide therapy within the bone marrow from a total of 39 transfusion-dependent patients with low- or intermediate-1-risk MDS with del(5q) chromosome aberration (MDS.5q-) who were recruited and treated with lenalidomide (10 mg / d) at the European site of the MDS-003 study and whose bone marrow samples (aspirates and biopsies) were taken prospectively at 6-month intervals after start of treatment. Mature erythroid precursors were marked by anti-hemoglobin-A-antibody (HbA); immature, HbA- erythroid precursors by anti-glycophorin-C-antibody (GPC), and the SC compartment by anti-CD34 antibody. Due to their low number, the numerical densities of SC-e were estimated indirectly by the numerical density of erythroblastic islands within bone marrow whereas the total number of SC was estimated by a statistical approach assuming that SC give rise to clusters of CD34+ precursors. The results were correlated with lenalidomide dose, RPS-14, SPARC, and glycophorin A (GPA) gene expression, degree of anemia, transfusion dependence, cytogenetic and molecular (FISH) response, clonal evolution, AML-free and overall survival of patients. RESULTS:Lenalidomide therapy resulted in a marked increase of SC-e, differentiation of SC-e to GPC+HbA- erythroid precursors, pronounced production of HbA+ precursors and erythrocytes resulting in transfusion independence in 54 % of patients. Lenalidomide-induced increase of SC-e did not correlate with a rise in the total number of SC. It was more pronounced in patients with a cytogenetic response, but it occurred also in patients without a cytogenetic remission. 17 / 39 patients with a minimal therapy effect on erythropoiesis achieving the 1% percentile of normal healthy marrow and continuing for > 12 months showed an excellent prognosis with a probability of survival > 90 % 60 months after start of treatment and 120 months after diagnosis of disease irrespective of their hematologic response whereas the median survival time of the patients without minimal effect on erythroid precursors (n = 22) was shorter than 20 months after start of treatment (P < 0.0002). In multivariate survival analysis, changes of erythropoiesis occurring during lenalidomide treatment, clonal evolution of disease, and age of patients were the only variables providing independent prognostic information (P < 0.00005). Classification of disease (FAB, WHO, IPSS, WPSS), changes of peripheral blood cell counts, transfusion independence, cytogenetic and molecular response, RPS-13, SPARC, or GPA gene expression did not provide additional independent prognostic information (P > 0.05). CONCLUSIONS:Lenalidomide promotes erythroid differentiation at stem cell level as well as at more mature stages of erythropoiesis affecting both, the neoplastic clone as well as non-neoplastic hematopoiesis. The anti-neoplastic effect of lenalidomide does not seem to be relevant for the prognosis of the patients; (1) genetic instability of neoplastic SC / precursor resulting in clonal evolution on the one and (2) the capacity of neoplastic SC / precursors to respond to lenalidomide therapy with the production of SC-e and more mature erythroid precursors on the other hand appear to be significant. By combining both variables, three groups can be discriminated allowing for a therapy-specific prognostic scoring: one group with excellent prognosis (100 % survival 5 years after start of therapy) comprising ∼ 33 % of patients, another with poor prognosis (0 % survival 5 years after start of therapy) comprising ∼ 28 % of patients, and a third group with intermediate prognosis (53 % dying during this period of observation) comprising ∼ 39 % of patients (P < 0.000001). Disclosures:Buesche:Celgene Corp.: reference pathologist of the MDS-004 and the MDS-Le-Mon-5 studies. Giagounidis:Celgene Corp: Consultancy, Honoraria, Membership on an entity’s Board of Directors or advisory committees. Göhring:Celgene Corp.: reference cytogeneticist of the MDS-004 study. Schlegelberger:Celgene Corp.: reference cytogeneticist of the MDS-004 study. Knight:Celgene Corp.: Employment.

Study with positron emission tomography of the osmotic opening of the dog blood-brain barrier for quinidine and morphine.

A canine model was used to evaluate the possibilities offered by positron emission tomography (PET) for the study of drug distribution in the brain during altered states of the blood-brain barrier (BBB). PET was used to monitor the changes in the distribution of [11C]quinidine and [11C]morphine resulting from BBB-disruption by intracarotid infusion of a hyperosmolar mannitol solution. Injection of Evans blue dye allowing post-mortem evaluation of the degree of BBB-opening was used as a reference method. Brain radioactivity concentrations observed after i.v. injection of either [11C] quinidine or [11C]morphine were markedly increased by intracarotid mannitol infusion, whereas they were not affected by saline infusion. For both drugs a close correlation was found between the radioactivity concentrations and the degree of Evans blue staining within the brain hemispheres and within smaller regions of interest corresponding to quadrants of a hemisphere. This parallelism between the findings for radioactivity concentrations and Evans blue staining suggests that PET allows the detection of in-vivo changes in brain distribution of drugs resulting from alterations of the BBB permeability.

Prolonged mechanical and physiological changes in the upper extremity following short-term simulated power hand tool use

This study investigated in-vivo changes in upper limb dynamic mechanical properties and magnetic resonance imaging (MRI) parameters following short-term power hand tool operation. Previous studies have found reduction in mechanical properties following short-term power tool usage at long build-up times. This study advances that work by having participants operate a simulated pistol grip power hand tool and evaluating changes in mechanical properties, strength, discomfort level and MRI prior to tool operation and daily for 3 d after tool operation. Twenty-four participants were randomly assigned to operate a simulated power hand tool for either a high peak reaction force of 123 N (peak torque = 8 Nm, build-up time = 250 ms) or at a low peak reaction force of 5 N (peak torque = 2 Nm, build-up time = 50 ms). Subjects operated the tool for 60 min at the rate of six times per min. A reduction in stiffness (27%, p < 0.05) was observed 24 h after tool operation for the high force group and this change persisted (26%, p < 0.05) up to 72 h after tool operation. Similar changes were not observed for the low force group. No changes were observed in mass moment of inertia, damping, isometric strength and damping for either group (p > 0.05). There was a signal intensity increase (12%, CI 19%, 5.06%) in the supinator muscle MRI for both groups 24 h after tool operation but only the high force group remained elevated (10%, CI 13.7%, 0.06%) 72 h after tool operation. Persistent short-term changes in mechanical and MRI parameters at high force levels could indicate increased strain on the upper limb and may negatively affect ability to react during rapid forceful loading of the upper limb. This research can ultimately lead to better ergonomic interventions through quantitative power hand tool design guidelines and work practices based on understanding the damaging effects of exposure to specific levels of reaction force, build-up time and repetition, as well as providing new outcome measures for epidemiological studies.

Effect of diet and exercise on serum insulin, IGF-I, and IGFBP-1 levels and growth of LNCaP cells in vitro (United States).

Accumulating evidence indicates that prostate cancer is associated with high levels of serum IGF-I. This study was conducted to determine whether a low-fat diet and exercise (DE) intervention may modulate the IGF axis and reduce prostate cancer cell growth in vitro. Fasting serum was obtained from 14 men (age 60 +/- 3 years) participating in an 11-day DE program and from eight similarly aged men who had followed the DE program for 14.2 +/- 1.7 years (long-term). Insulin, IGF-I, IGFBP-1, and IGFBP-3 were measured by ELISA, and serum was used to stimulate LNCaP cell growth in vitro. Serum IGF-I levels decreased by 20% while IGFBP-1 increased by 53% after 11-day DE. In the long-term group, IGF-I was 55% lower, while IGFBP-1 was 150% higher relative to baseline. Serum insulin decreased by 25% after 11-day DE and was 68% lower in the long-term group, relative to baseline. No changes in serum IGFBP-3 were observed. Serum-stimulated LNCaP cell growth was reduced by 30% in post-11-day serum and by 44% in long-term serum relative to baseline. LNCaP cells incubated with post-DE serum showed increased apoptosis/ necrosis, compared to baseline. A low-fat diet and exercise intervention induces in-vivo changes in the circulating IGF axis and is associated with reduced growth and enhanced apoptosis/necrosis of LNCaP tumor cells in vitro.

Comparison of early onco/suppressor gene expressions in peripheral leukocytes and potential target organs of rats exposed to the carcinogen 1-nitropyrene.

An in-vivo model has been developed to study early expressions of c-myc, Ha-ras oncogenes and p53 suppressor gene as biomarkers of carcinogenic exposure and/or tumorigenesis. In order to validate the in-vivo expression changes as biomarkers, rats were treated with the outdoor air pollutant carcinogen 1-nitropyrene. The gene expression levels were measured after 24 and 48 h in potential target tissues (lung, liver, lymph nodes, kidneys, spleen) and in peripheral blood leukocytes. Another main objective was to prove the applicability of leukocytes as a surrogate tissue, having a similar expression pattern of the selected genes upon carcinogenic exposure. The c-myc oncogene was not suitable as an early biomarker because of the lack or low level of its expression. However, in the case of the other oncogene Ha-ras and the suppressor gene p53, remarkable and early changes were detected in the expression signals. Similar expression patterns could only be detected in leukocytes and the spleen; therefore we continue this validation study by using other types and routes of exposure.

LOCAL COLLAGEN-TURNOVER IN HUMAN CONNECTIVE TISSUE AFTER STRENUOUS EXERCISE.

1857 Animal studies have shown that physical training increases the ultimate strength of tendons possibly explained by accelerated collagen synthesis (Suominen 1980). Markers of collagen turnover have been shown to increase in plasma in response to exercise (Virtanen 1993, Eriksen 1995). In the present study the local turnover of collagen was measured as release of PICP (marker of collagen type I synthesis) and ICTP (marker of collagen type I breakdown) in the peritendinous space of the Achilles tendon by microdialysis before, immediatly after and 72 hours after strenuous exercise (3 hours running, 12 km/h) in 7 long distance runners (6 M, 1 F). In addition concentrations of PICP and ICTP were measured in the arterial blood. From dialysate concentrations and determined recovery by the internal reference method, interstitial concentration of PICP (19 ± 3 ng/ml) was found to be unchanged immediately after exercise (P > 0,05), whereas a 3 fold increase in the local PICP concentration was found 72 hours after the run (51 ± 18 ng/ml) (P < 0,05). No change in the arterial PICP-concentration was found. ICTP-level was found to be unchanged both in tissue and serum (P > 0,05). The study demonstrates that the microdialysis technique enables monitoring of the in-vivo changes of collagen-turnover locally in the area around the Achilles tendon. It was found that 72 hours following a 3 hour-run the concentration of PICP increases significantly. These findings indicate that exercise stimulates synthesis of collagen type I in human tendon.

Biomechanical model for the myogenic response in the microcirculation: Part II--Experimental evaluation in rat cremaster muscle.

In order to test experimentally a novel model for myogenic contraction of the arterioles, presented in Part I of this series, a sequence of in-vivo studies was performed on arterioles of rat cremaster muscle using a modified "Box Method." The tissue was enclosed in a sealed chamber in which the extravascular pressure could be changed as a means of stimulating the myogenic response. The microvascular pressures were measured using an improved cremaster preparation with intact distal feeder. The experiment consists of measurements of a static myogenic response and the response to a step extravascular pressure, as well as passive viscoelastic properties of the arteriole. These measurements served to determine the parameters involved in the theoretical model. The model prediction were then compared with in-vivo observations during a ramp and during oscillatory extravascular pressure changes in the same arterioles. The results indicate that the model is capable of quantitatively predicting time dependent in-vivo changes in response to transmural pressure. The measured model parameters suggest an increase in myogenic activity from proximal arcade arterioles to more distal transverse arterioles in cremaster muscle of Wistar rats.