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Abstract Objective Avoiding abacavir in HIV-infected patients tested positive for HLA-B*5701 reduces the risk of abacavir hypersensitivity reaction (ABC-HSR). Our aim was to assess the costs of clinically suspected HSR and to estimate potential cost savings of implementing prospective HLA-B*5701-screening for HIV-infected patients initiating abacavir/lamivudine fixed-dose combination (ABC/3TC FDC) compared to initiating respective treatment without screening. Methods Employing a decision tree model the expected HSR-related costs of screening vs. no screening were estimated from the societal and healthcare payer perspective (reference year 2007). A retrospective standardized assessment of all clinically suspected ABC-HSR cases without screening at 5 German HIV-centres was performed to measure resource consumption. In- and outpatient care, discarded ABC/3TC FDC and concomitant medication were considered. Direct resource utilization was valued using German fees (EBM, G-DRGs). Indirect costs were measured with the human capital approach. Estimates for the HLA-B*5701-prevalence, HSR-incidence, and hospitalization rate were based on clinical trials and cohorts and it was assumed that screening reduces the incidence of clinically suspected ABC-HSR from 10% to 0.5%. Results Thirty-two ABC-HSR cases were identified from 1998 to 2007. Mean direct and total costs per clinically suspected HSR case were € 1,362 and € 2,235, respectively. Hospital costs contributed 63.3% to direct costs. Potential cost savings when implementing genetic screening were estimated at € 44 and € 127 per screened patient, from a healthcare payer or societal perspective. Conclusion HLA-B*5701 screening prior to ABC/3TC FDC initiation prevents significant HSR-related costs per screened patient and is likely to lead to overall net savings.

Respiratory diseases like pulmonary arterial hypertension (PAH) frequently exhibit sexual dimorphism. Female PAH patients are more susceptible to the disease but have increased survival rates. This phenomenon is known as the estrogen paradox, and the underlying mechanisms are not fully understood. During PAH progression in vivo, human pulmonary arterial adventitial fibroblasts (hPAAFs) differentiate into an activated phenotype. These cells produce excess, aberrant extracellular matrix proteins that stiffen the surrounding pulmonary arterial tissues. Here, we employed dynamic poly(ethylene glycol)-alpha methacrylate (PEGαMA)-based biomaterials to study how the age and sex of human serum influenced hPAAF activation in response to microenvironmental stiffening in vitro . Results showed female and male cells responded differently to increases in microenvironmental stiffness and serum composition. Male hPAAFs were less activated than female cells on soft hydrogels and more responsive to increases in microenvironmental stiffness regardless of serum composition. Female hPAAF activation followed this pattern only when cultured in younger (age < 50) female serum or when older (age ≥ 50) female serum was supplemented with estradiol. Otherwise, female hPAAF activation was relatively high on both soft and stiffened hydrogels, with little difference in activation between the two conditions. Collectively, these results suggest that it may be possible to model the estrogen paradox observed in PAH in vitro and that it is critical for researchers to report cell sex and serum source when conducting in vitro experimentation., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Chelsea M. Magin reports financial support was provided by University of Colorado Anschutz Medical Campus Ludeman Family Center for Women’s Health Research. Chelsea M. Magin reports financial support was provided by Rose Community Foundation. Chelsea M. Magin reports financial support was provided by National Heart Lung and Blood Institute. Chelsea M. Magin reports financial support was provided by National Science Foundation. Chelsea M. Magin reports financial support was provided by Colorado Pulmonary Vascular Disease Research Award. Mikala C. Mueller reports financial support was provided by National Center for Advancing Translational Sciences Clinical and Translational Science Awards Program. Chelsea M. Magin reports a relationship with Colorado Bioscience Institute that includes: board membership. Chelsea M. Magin reports a relationship with Boulder IQ that includes: consulting or advisory. Chelsea M. Magin has patent #PCT/US2019/012722 pending to University of Colorado. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

Because people living with HIV (PLWH) have an elevated risk for cardiovascular disease (CVD), prevention of CVD should be integrated in to HIV care. In this study, we compared the agreement between three risk scores and evaluated the indication for statin therapy based on guidelines of the American Heart Association and European AIDS Clinical Society. This study is a cross-sectional, single-center study. All PLWH ≥ 30 years without CVD and statin therapy were consecutively enrolled. Agreement between CVD risk estimates was assessed using Cohen's kappa coefficient. Of 488 PLWH, 41.2% were female with a median age of 47.8 years. D:A:D-R classified the highest proportion of patients in the categories of high/very high risk for CVD (17.8%) compared to SCORE (4.7%) and FRS (13.7%). D:A:D-R and SCORE (κ = 0.11) as well as D:A:D-R and FRS (κ = 0.33) showed poor agreement. Based on different CVD risk equations and guidelines, indication for statin therapy ranged from 34.8% to 92.0% of patients. In conclusion, a high proportion of PLWH is at high risk for CVD likely underestimated by treating physicians. Inconsistencies in the evaluation of CVD risk and primary prophylaxis should be tackled by an interdisciplinary approach.

Background: Cytomegalovirus (CMV) infection is one of the main driving forces of T-cell senescence in the general population, whereas its differential impact in people living with HIV (PLWH) is less well characterized. The study explores the effect of latent CMV infection on T-cell subsets, monocyte/macrophages activation markers, and CRP in PLWH on long-term ART., Methods: Cross-sectional cohort study including PLWH on long-term suppressive ART. Individuals of 4 groups (HIV+CMV-, HIV+CMV+, HIV-CMV+, and HIV-CMV-) were matched 1:1:1:1 for age and sex. Immunophenotyping of lymphocyte and T-cell subsets by multicolor flow cytometry was performed in fresh blood samples collected from patients and healthy donors., Results: Both, latent CMV and treated HIV infection were associated with an expansion of CD8 T cells, a reduced CD4/CD8 ratio, and with CD8 T-cell activation with a cumulative effect in CMV/HIV-coinfected individuals. CMV was associated with elevated numbers of late effector and terminally differentiated CD8 T-cells. Compared to CMV monoinfection, CMV/HIV coinfection showed to be associated with lower proportion of CD28-CD8+ T cells expressing CD57 suggesting that HIV preferentially expands CD28-CD57-CD8+ T cells and impedes terminal differentiation of CD28-CD8+ T cells. We could not show any association between HIV or CMV infection status and concentration of CRP and CD163., Conclusions: CMV infection is associated with phenotypic signs of T-cell senescence, promoting exacerbation and persistence of alterations of the T-cell compartment in PLWH on effective ART, which are associated with adverse clinical outcomes and may be an attractive target for therapeutic interventions.

Ecology and epidemiology of Echinococcus multilocularis and human alveolar echinococcosis (AE) are changing in Central Europe. Our data from a regional referral center for AE in southwest Germany suggest rising regional incidence for AE (annual incidence per 100,000 population 2004-2011: 0.12; 2012-2019: 0.20) and emerging urban AE (of 7 cases of AE in Freiburg city dwellers none was diagnosed before 2012) calling for an intensification of E. multilocularis and AE surveillance and of AE prevention measures.

Introduction: Data on people living with human immunodeficiency virus (PLWH) in the current SARS-CoV-2 pandemic are still scarce. This case series of 33 PLWH patients with COVID-19 reveals symptoms and outcome in this special population., Methods: Retrospective analysis of anonymized data including age, gender, HIV-associated parameters, symptoms, and outcome., Results: Three out of 32 patients with documented outcomes died (9%). 91% of the patients recovered and 76% have been classified as mild cases. All patients were on antiretroviral treatment, of them 22 on tenofovir-containing regimen and 4 on the protease inhibitor darunavir., Conclusions: This preliminary case series does not support excess morbidity and mortality among symptomatic COVID-19 PLWH and with viral suppression on ART. SARS-CoV-2 infections may occur during boosted darunavir-based and/or on tenofovir-containing ART.

Minimal residual disease (MRD) is a powerful prognostic factor in acute lymphoblastic leukemia (ALL) and is used for patient stratification and treatment decisions, but its precise role in Philadelphia chromosome positive ALL is less clear. This uncertainty results largely from methodological differences relating to the use of real-time quantitative PCR (qRT-PCR) to measure BCR-ABL1 transcript levels for MRD analysis. We here describe the first results by the EURO-MRD consortium on standardization of qRT-PCR for the e1a2 BCR-ABL1 transcript in Ph + ALL, designed to overcome the lack of standardisation of laboratory procedures and data interpretation. Standardised use of EAC primer/probe sets and of centrally prepared plasmid standards had the greatest impact on reducing interlaboratory variability. In QC1 the proportion of analyses with BCR-ABL1/ABL1 ratios within half a log difference were 40/67 (60%) and 52/67 (78%) at 10 -3 and 36/67 (53%) and 53/67 (79%) at 10 -4 BCR-ABL1/ABL1. Standardized RNA extraction, cDNA synthesis and cycler platforms did not improve results further, whereas stringent application of technical criteria for assay quality and uniform criteria for data interpretation and reporting were essential. We provide detailed laboratory recommendations for the standardized MRD analysis in routine diagnostic settings and in multicenter clinical trials for Ph + ALL.

Few studies have investigated skin blood flow in children and age-related differences in the underlying mechanisms. We examined mechanisms of skin blood flow responses to local heating, postocclusive reactive hyperaemia (PORH), and isometric handgrip exercise in adult and prepubescent males, hypothesizing that skin blood flow responses would be greater in children compared with adults. We measured skin blood flow in 12 boys (age, 9 ± 1 years) and 12 men (age, 21 ± 1 years) using laser-Doppler flowmetry at rest, in response to 3-min PORH, 2-min isometric handgrip exercise, and local skin heating to 39 °C (submaximal) and 44 °C (maximal). Using wavelet analysis we assessed endothelial, neural, and myogenic activities. At rest and in response to local heating to 39 °C, children had higher skin blood flow and endothelial activity compared with men (d ≥ 1.1, p < 0.001) and similar neurogenic and myogenic activities (d < 0.2, p > 0.05). Maximal responses to 44 °C local skin heating, PORH, and isometric handgrip exercise did not differ between boys and men (all d ≤ 0.2, p > 0.05). During PORH children demonstrated greater endothelial activity compared with men (d ≥ 0.6, p < 0.05); in contrast, men had higher neurogenic activity (d = 1.0, p < 0.01). During isometric handgrip exercise there were no differences in endothelial, neurogenic, and myogenic activities (d < 0.2, p > 0.3), with boys and men demonstrating similar increases in endothelial activity and decreases in myogenic activity (d ≥ 0.8, p < 0.05). These data suggest that boys experience greater levels of skin blood flow at rest and in response to submaximal local heating compared with men, while maximal responses appear to be similar. Additionally, endothelial mediators seem to contribute more to vasodilatation in boys than in men.

Genomic events associated with poor outcome in chronic myeloid leukemia (CML) are poorly understood. We performed whole-exome sequencing, copy-number variation, and/or RNA sequencing for 65 patients to discover mutations at diagnosis and blast crisis (BC). Forty-six patients with chronic-phase disease with the extremes of outcome were studied at diagnosis. Cancer gene variants were detected in 15 (56%) of 27 patients with subsequent BC or poor outcome and in 3 (16%) of 19 optimal responders ( P = .007). Frequently mutated genes at diagnosis were ASXL1 , IKZF1 , and RUNX1 The methyltransferase SETD1B was a novel recurrently mutated gene. A novel class of variant associated with the Philadelphia (Ph) translocation was detected at diagnosis in 11 (24%) of 46 patients comprising fusions and/or rearrangement of genes on the translocated chromosomes, with evidence of fragmentation, inversion, and imperfect sequence reassembly. These were more frequent at diagnosis in patients with poor outcome: 9 (33%) of 27 vs 2 (11%) of 19 optimal responders ( P = .07). Thirty-nine patients were tested at BC, and all had cancer gene variants, including ABL1 kinase domain mutations in 58%. However, ABL1 mutations cooccurred with other mutated cancer genes in 89% of cases, and these predated ABL1 mutations in 62% of evaluable patients. Gene fusions not associated with the Ph translocation occurred in 42% of patients at BC and commonly involved fusion partners that were known cancer genes (78%). Genomic analysis revealed numerous relevant variants at diagnosis in patients with poor outcome and all patients at BC. Future refined biomarker testing of specific variants will likely provide prognostic information to facilitate a risk-adapted therapeutic approach., (© 2018 by The American Society of Hematology.)

High mortality rates of invasive fungal disease (IFD), especially invasive aspergillosis (IA), in immunocompromised haematological patients and current diagnostic limitations require improvement of detection of fungal pathogens by defining the optimal use of biomarkers and clinical samples. Concurrent bronchoalveolar lavage (BAL) and peripheral blood samples of 99 haematological patients with suspected IFD were investigated within a multicentre prospective study. Diagnostic performance of a galactomannan (GM) enzyme immune assay (EIA), a 1,3-β-D-glucan assay (BDG), an Aspergillus PCR, and a multifungal DNA-microarray (Chip) alone or in combination were calculated. IFD were classified as proven (n=3), probable (n=34), possible (n=33), and no IFD (n=29) according to EORTC/MSG criteria. GM, PCR, and Chip showed superior diagnostic performance in BAL than in blood, whereas specificity of BDG in BAL was poor (48% (14/29)). The combination of GM (BAL) with BDG (blood) showed sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and DOR (diagnostic odds ratio) of 92% (34/37), 93% (27/29), 94%, 90%, and 153.0, respectively. Combining GM (BAL) with PCR (BAL) showed convincing diagnostic potential for diagnosing IA with sensitivity, specificity, PPV, NPV, and DOR of 85% (17/20), 97% (28/29), 94%, 90%, and 158.7. Addition of the DNA-microarray resulted in further detection of two mucormycetes infections. In 1 out of 15 Aspergillus DNA-positive samples a triazole resistance-mediating Cyp51A mutation was found. Combination of biomarkers is superior to their sole use in diagnosing IFD, particularly IA. Integrating blood and BAL samples into a diagnostic algorithm is an advantageous approach., (Copyright © 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: The investigation of ischemic stroke etiology is commonly limited to the heart and extracranial vessels. Nevertheless, the diagnosis of intracranial stenosis may carry important therapeutic implications. The aims of this study were to determine the prevalence and clinical predictors of intracranial atherosclerotic stenosis (ICAS) in a sample of patients with ischemic stroke., Methods: Consecutive patients admitted to a university-based outpatient stroke clinic underwent CT angiography of the intracranial and extracranial brain vessels. Clinical, demographic, and laboratory characteristics were compared between patients with increasing levels of stenosis. Ankle-brachial index (ABI) was measured to quantify peripheral arterial disease, defined as an ABI less than or equal to .9. Multivariable ordinal logistic regression was constructed to predict increasing stenosis grades (none, 1%-49%-mild, 50%-69%-moderate, 70%-100%-severe)., Results: We studied 106 subjects, mean age 62 ± 15 years, 54% female. ICAS was present in 38 (36%) patients: 19 (50%) mild, 7 (18%) moderate, and 12 (32%) severe. Of 74 patients where ABI was measured, low ABI was found more frequently with increasing ICAS severity (26%, 42%, 67%, and 89% of patients with none, mild, moderate, and severe ICAS, respectively). In univariable analysis, higher age, presence of diabetes, abdominal obesity, and low ABI correlated with increasing stenosis grades. In multivariable analysis, only low ABI remained independently associated with increasing stenosis grades., Conclusions: The ABI is independently associated with increasing severity of ICAS, making it a potentially useful triaging tool for more invasive test selection., (Copyright © 2016 National Stroke Association. Published by Elsevier Inc. All rights reserved.)

Purpose: Germany is witnessing an increase in the number of new infections with human immunodeficiency virus (HIV). Enabling persons living with HIV (PLHIV) to adopt safer sex practices might contribute towards reducing the incidence of HIV infections. The aim of this study was to identify gaps in the sexual and reproductive health (SRH) services provided to PLHIV in Germany., Methods: Within the framework of the European public health project Eurosupport 5, self-reported questionnaires were distributed to PLHIV and a survey of SRH-service providers was carried out. The completed questionnaires and survey results were analysed., Results: Of the questionnaires distributed, 218 PLHIV (90 % men, 10 % women) returned a completed questionnaire. Of these, 74 % self-identified as men having sex with men (MSM) and 13 % as heterosexual men. MSM reported a median number of ten casual partners in the previous 6 months and unprotected sex in one-third of anal intercourses with casual partners, demonstrating that this group adopted more risky sexual behaviours than heterosexual PLHIV. Even though all PLHIV stated they would appreciate more support and service providers indicated that they provided a wide range of SRH services, SRH-relevant topics were rarely discussed between PLHIV and service providers. According to the patients' perception, shortage of time, lack of initiative by service providers and their own difficulty to address SRH-related topics were the most relevant obstacles to receiving satisfactory support., Conclusion: Many PLHIV consult their HIV-physician regularly for medical follow-up and also indicate that HIV-physicians should be the source of information concerning SRH counselling. HIV-physicians should take advantage of their key role in HIV care and strengthen their efforts to integrate SRH services in routine HIV care.

How reproductive isolation is related to divergent natural selection is a central question in speciation. Here, we focus on several ecologically specialized taxa or 'call types' of red crossbills (Loxia curvirostra complex), one of the few groups of birds providing much evidence for ecological speciation. Call types differ in bill sizes and feeding capabilities, and also differ in vocalizations, such that contact calls provide information on crossbill phenotype. We found that two call types of red crossbills were more likely to approach playbacks of their own call type than those of heterotypics, and that their propensity to approach heterotypics decreased with increasing divergence in bill size. Although call similarity also decreased with increasing divergence in bill size, comparisons of responses to familiar versus unfamiliar call types indicate that the decrease in the propensity to approach heterotypics with increasing divergence in bill size was a learned response, and not a by-product of calls diverging pleiotropically as bill size diverged. Because crossbills choose mates while in flocks, assortative flocking could lead indirectly to assortative mating as a by-product. These patterns of association therefore provide a mechanism by which increasing divergent selection can lead to increasing reproductive isolation.

Background: Targeted treatment of chronic myelogenous leukemia using imatinib has dramatically improved patient outcome. However, residual disease can be detected in the majority of patients treated with imatinib. Compensatory activation of MAP kinases (MAPK1/2) in response to BCR-ABL-inhibitors has been reported as a potential cytokine-dependent resistance mechanism leading to the rescue of leukemic progenitor cells., Methods: Differential MAPK-modulating activity of clinically approved tyrosine kinase inhibitors was assessed in vitro using BCR-ABL-transformed cells. CD34+-enriched progenitors of newly diagnosed chronic myelogenous leukemia patients were exposed to tyrosine kinase inhibitors. MAPK-signaling was studied by Western blot technique. Proliferation assays were used to analyze response to antileukemic treatment., Results: The ABL-inhibitors imatinib and nilotinib activate MAPKs in CD34+ chronic myelogenous leukemia progenitor cells, whereas treatment with the SRC/ABL-inhibitor dasatinib does not affect MAPK-activation at clinically relevant concentrations. Similar results are seen in BCR-ABL-transformed cells in the presence of interleukin-3 (IL-3). Experiments using BCR-ABL-mutant T315I, a resistance mutation not amenable to tyrosine kinase inhibitor binding, demonstrate that ABL-inhibitor-induced MAPK-activation does not depend on BCR-ABL-inhibition and cannot be prevented by selective SRC-inhibition. However, BCR-ABL-T315I enhances MAPK-activation, suggesting a T315I-dependent positive feedback of MAPK-activation. An autocrine IL-3-loop as trigger for aberrant T315I-dependent MAPK-activation was excluded., Conclusions: Aberrant MAPK-activation triggered by ABL-inhibitors and positively regulated by BCR-ABL kinase mutation T315I might be an experimental explanation for the clinical observation that patients carrying high-resistance mutations show a highly aggressive course of their disease when tyrosine kinase inhibitor treatment is not discontinued in time.

Objective: To assess forelimbs and hind limb joint kinematics in dogs during walking on an inclined slope (uphill), on a declined slope (downhill), or over low obstacles (cavaletti) on a horizontal surface and compare findings with data acquired during unimpeded walking on a horizontal surface., Animals: 8 nonlame dogs (mean +/- SD age, 3.4 +/- 2.0 years; weight, 23.6 +/- 4.6 kg)., Procedures: By use of 10 high-speed cameras and 10 reflecting markers located on the left forelimbs and hind limbs, joint kinematics were recorded for each dog during uphill walking, downhill walking, and walking over low obstacles or unimpeded on a horizontal surface. Each exercise was recorded 6 times (10 s/cycle); joint angulations, angle velocities and accelerations, and range of motion for shoulder, elbow, carpal, hip, stifle, and tarsal joints were calculated for comparison., Results: Compared with unimpeded walking, obstacle exercise significantly increased flexion of the elbow, carpal, stifle, and tarsal joints and extension in the carpal and stifle joints. Only uphill walking caused increased hip joint flexion and decreased stifle joint flexion; downhill walking caused less flexion of the hip joint. During obstacle exercise, forward angle velocities in the elbow and stifle joints and retrograde velocity in the tarsal joint changed significantly, compared with unimpeded walking. Joint angle acceleration of the elbow joint changed significantly during all 3 evaluated exercises., Conclusions and Clinical Relevance: These evidence-based data indicated that each evaluated exercise, except for downhill walking, has a specific therapeutic value in physical therapy for dogs.

Nilotinib is a highly selective Bcr-Abl inhibitor approved for imatinib-resistant chronic myeloid leukemia (CML). Nilotinib and dasatinib, a multi-targeted kinase inhibitor also approved for second-line therapy in CML, have different patterns of kinase selectivity, pharmacokinetics, and cell uptake and efflux properties, and thus patients may respond to one following failure of the other. An international phase II study of nilotinib was conducted in CML patients (39 chronic phase (CP), 21 accelerated phase (AP)) after failure of both imatinib and dasatinib. Median times from diagnosis of CP or AP to nilotinib therapy were 89 and 83 months, respectively. Complete hematological response and major cytogenetic response (MCyR) rates in CP were 79% and 43%, respectively. Of 17 evaluable patients with CML-AP, 5 (29%) had a confirmed hematological response and 2 (12%) a MCyR. The median time to progression has not yet been reached in CP patients. At 18 months 59% of patients were progression-free. Median overall survival for both populations has not been reached, and the estimated 18-month survival rate in CML-CP was 86% and that at 12 months for CML-AP was 80%. Nilotinib is an effective therapy in CML-CP and -AP following failure of both imatinib and dasatinib therapy.

A 31-year-old man with ankylosing spondylitis, receiving treatment with infliximab, presented with a large progressive cutaneous ulcer at the right knee. Biopsies showed Leishmania amastigotes, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis showed Leishmania infantum as the causative agent. After treatment with miltefosine, the ulcer resolved completely, and infliximab was reinstituted because of progression of spondylitis. After 1 year, there was a recurrent ulcer at the same site being positive for Leishmania DNA by PCR. Local treatment with sodium stibogluconate resulted in complete regression. Cutaneous leishmaniasis should be added to the list of opportunistic infections associated with anti-tumor necrosis factor (TNF) treatment. Despite recurrences, antileish-manial treatment may be effective in cases without alternatives to anti-TNF therapy.

Background and Aims: Aim of this study was to define the perioperative risk profile in surgery of patients suffering from Parkinson's disease (PD) in order to improve treatment options in these patients., Materials and Methods: Over a period of 13 years, 51 patients suffering from Parkinson's disease treated in the departments of general, visceral, thoracic, vascular, and trauma surgery were retrospectively compared using matched-pair analysis with 51 controls not affected by PD. Both groups of patients were assessed regarding morbidity and mortality, length of treatment, and rehabilitation., Results: Surgical patients suffering from Parkinson's disease showed an increase in risk of morbidity. Postoperative falls occurred more commonly (entire cohort, p < 0.03). In PD patients treated in the trauma surgery department, postoperative falls (p < 0.04), postoperative stay (p < 0.03), and overall duration of treatment (p < 0.02) were significantly longer than in patients without PD. PD patients of the trauma unit could be discharged home less often for ambulantory rehabilitation after in-patient treatment (p < 0.03)., Conclusions: Concomitant Parkinson's disease is a significant factor of perioperative morbidity in surgical patients, especially of patients treated in the trauma unit. Perioperative morbidity in PD patients may be influenced by early diagnostic and therapeutic measures.

Objectives: In this report two cases of partial gastrocnemius muscle avulsion treated with pulsed therapeutic ultrasound are described., Methods: The outcome in these two dogs was evaluated using ultrasonographic imaging and the measurement of ground reaction forces with a force plate., Results: Both dogs showed an amelioration of the clinical signs within one month after commencement of the ultrasound therapy. The follow-up time for these cases was one year and six months respectively. Both of the dogs were free of lameness and had returned to their normal amount of exercise. Palpation of the fabella associated with the muscle injury did not produce any signs of pain. Ultrasonographic imaging did not detect any signs of haemorrhage or oedema, although scarring of muscle fibres was present. The force-plate analyses revealed an improvement., Clinical Significance: These results suggest that therapeutic ultrasound could be a beneficial treatment modality for this kind of muscle injury.

Actual BCR-ABL kinase inhibition in vivo as determined by phospho-CRKL (pCRKL) monitoring has been recognized as a prognostic parameter in patients with chronic myelogenous leukemia treated with imatinib. We report a biomarker sub-study of the international phase I clinical trial of nilotinib (AMN107) using the established pCRKL assay in imatinib-resistant chronic myeloid leukemia or Ph+ acute lymphoblastic leukemia. A minimum dose (200 mg) required for effective BCR-ABL inhibition in imatinib resistant/intolerant leukemia was determined. The pre-clinical activity profile of nilotinib against mutant BCR-ABL was largely confirmed. Substantial differences between peripheral blood baseline pCRKL/CRKL ratios were observed when comparing chronic myeloid leukemia with Ph+ acute lymphoblastic leukemia. Finally, rapid BCR-ABL-reactivation shortly after starting nilotinib treatment was seen in acute lymphoblastic leukemia patients with progressive disease carrying the P-loop mutations Y253H, E255K, or mutation T315I. Monitoring the actual BCR-ABL inhibition in nilotinib treated patients using pCRKL as a surrogate is a means to establish effective dosing and to characterize resistance mechanisms against nilotinib.

A 59-year-old, healthy Croatian presented with a slowly growing tumor in the left lower abdomen, which was slightly painful on compression. He complained of neither dyspepsia nor fever. There were no pathologic findings in laboratory analysis, particularly no elevation of leukocytes or C-reactive protein. MRI of the abdomen (T1w, fat saturated, and iv-contrast) shows a diffuse contrast enhancing mass of the left abdominal wall (Figure 1a, arrow) with infiltration of the peritoneal cavity (Figure 1b, arrow). Because a malignant process was suspected the patient underwent abdominal surgery and excision of the tumor. Histopathological examination showed chronic-fibrosing and granulocytic, abscess-forming inflammation with Gram- and PAS-positive bacteria, corresponding to the diagnosis of chronic actinomycosis (Figure 1c). Following surgery, the patient was treated 1 month with iv and 6 more months with oral penicillin. The patient remained well 1 year after surgery. Actinomycosis is a rare, chronic granulomatous disease, which affects most commonly the cervicofacial and abdominal area. Actinomycetes are filamentous, gram-positive, anaerobic bacteria and commensal inhabitants of the oral cavity and intestinal tract; however, they acquire pathogenicity through invasion of the breached tissue. Because of its rarity and non-specific symptoms, abdominal actinomycosis is usually diagnosed postoperatively since most patients undergo exploratory laparotomy for a suspected neoplasm.

Background: Recently, an International Scale was proposed for standardizing BCR-ABL transcript measurements and reporting in the assessment of minimal residual disease by real-time quantitative polymerase chain reaction (RQ-PCR). Here we present the setting up of the International Scale conversion factors for a national laboratory by performing both a cross-analysis of a set of standard samples from a reference laboratory and an analysis of bone marrow and peripheral blood samples at diagnosis (from 32 and 27 patients, respectively)., Design and Methods: A total of 222 bone marrow and 173 peripheral blood mononuclear cell samples from 96 patients with chronic myeloid leukemia were analyzed with RQ-PCR according to Europe Against Cancer protocols. Additionally, 291 bone marrow samples were analyzed with high mitotic index metaphase fluorescence in situ hybridization (metaphase FISH)., Results: Major molecular response according to the International Scale in BCR-ABL/GUS transcript levels corresponded to a ratio of 0.035% in peripheral blood and 0.034% in bone marrow, yielding the same conversion factor of 2.86 for both types of sample. Based on metaphase FISH, values of 10%/-1.0 log, 1%/-2.0 log and 0.1%/-3.0 log on the International Scale, corresponded to 13%, 2%, and 0.3% of Philadelphia chromosome positive cells in bone marrow, respectively., Conclusions: In conclusion, conversion factors can be determined either by cross-analyzing a number of samples with a laboratory that has already established the International Scale or utilizing sufficient numbers of reference samples from chronic myeloid leukemia patients at diagnosis, or using the upcoming international standards.

The concept of CCR5 antagonists introduces an additional molecular target. Maraviroc (MVR) is approved by the FDA for use in HIV-1 infected patients for combination antiretroviral treatment of adults infected with only CCR5-tropic HIV-1 who have evidence of viral replication and HIV-1 strains resistant to multiple antiretroviral agents. Tropism and treatment history should guide the use of MVR. Data from clinical trials show significant efficacy of MVR for patients with pre-treatment and multiple class failure. Additional clinical data show a CD4 reconstitution that is more pronounced than with comparator in treatment naive and in late stage patients even without CCR5-tropic virus indicating patients in earlier stages and even patients without CCR5 testing will benefit from MVR. MVR is not licensed for treatment naive patients but it has a high potential for further development in this patient group. It shows better immunological reconstitution than efavirenz. Pooled safety data from all available trials shows good short term tolerability. Caution is needed in hepatitis co-infection with pre-existing liver damage and in patients with heart failure. Isolates from different geographic regions differ in coreceptor usage. Summarizing knowledge on HIV-1 subtypes and CCR5 tropism shows that in principle all subtypes are susceptible to MVR. However, in subtypes A and D dualtropic and alternative coreceptor use were found. Clinical efficacy in patients from regions with A and D predominance should be studied in future trials. In conclusion, MVR will be of benefit for patients in various treatment situations and regions.

The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs, consisting of human preadipocytes and fibrin glue as a carrier matrix, requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells. Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors, such as VEGF and bFGF. The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a VEGF-vector. Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector. Transfection efficiency (GFP expression) and VEGF expression were determined in vitro by FACS analysis and VEGF immunoassay, respectively. In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane (CAM). Four million VEGF transfected cells were transferred within a fibrin matrix onto the CAM on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program. Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12% and 41% of surviving cells. Results of the VEGF immunoassay demonstrated that VEGF expression was significantly higher following transfection. Investigations on the CAM outlined a significantly higher rate of vascularization in the transfected vs. control population. Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a VEGF vector. The enhanced VEGF expression on transfected cells results in an increase of vascularization of the cell constructs on the CAM.

The advent of the Bcr-Abl selective tyrosine kinase inhibitor imatinib mesylate (Glivec, Gleevec, Novartis, East Hanover, NJ) has substantially changed the treatment landscape for chronic myelogenous leukemia (CML). However, some patients, primarily those with advanced disease, are either initially refractory to imatinib or eventually develop imatinib resistance. Imatinib resistance or intolerance frequently depends on the re-emergence of Bcr-Abl kinase activity, but can also indicate Bcr-Abl-independent disease progression. Results from phase II/III trials suggest rates of resistance and relapse correlate with stage of disease and with the monitoring parameters: hematologic, cytogenetic, and molecular responses. To date, more than 40 different point mutations that code for distinct single amino acid substitutions in the Bcr-Abl kinase domain have been isolated from imatinib-resistant patients. These mutations affect amino acids involved in imatinib binding or in regulatory regions of the Bcr-Abl kinase domain, resulting in decreased sensitivity to imatinib while retaining aberrant kinase activity. Early mutation detection may aid in risk stratification and molecular-based treatment decisions. To overcome imatinib-resistant disease, novel tyrosine kinase inhibitors with activity against imatinib-resistant mutations and/or with inhibition of alternative pathways, such as Src activation, have recently been developed. Additional strategies include imatinib dose escalation, combination therapy, and treatment interruption to stop clonal selection of resistant cells.

Treatment of chronic myeloid leukemia (CML) with the tyrosine kinase inhibitor imatinib represents a successful application of molecularly targeted cancer therapy. A rapid hematologic and cytogenetic response can be induced in the majority of people, even in advanced disease. However, complete eradication of malignant cells, which are characterized by the expression of the BCR-ABL1 fusion protein, is rare. Reasons for the persistence of the malignant clone are currently not known and provide a substantial challenge for clinicians and biologists. Based on a mathematical modeling approach that quantitatively explains a broad range of phenomena, we show for two independent datasets that clinically observed BCR-ABL1 transcript dynamics during imatinib treatment of CML can consistently be explained by a selective functional effect of imatinib on proliferative leukemia stem cells. Our results suggest the general potential of imatinib to induce a complete elimination of the malignant clone. Moreover, we predict that the therapeutic benefit of imatinib can, under certain circumstances, be accelerated by combination with proliferation-stimulating treatment strategies.

Successful augmentation of soft tissues by transplantation of preadipocytes within a matrix requires the formation of a new capillary network with connection to the host vessel system. Particularly, cells located centrally within the transplanted cell-matrix-construct represent a population with a blood supply questionable for survival. We demonstrated that under in vivo conditions preadipocytes possess the ability to induce and support the vascularization of the implant presumably by expression of several growth factors, such as VEGF (vascular endothelial growth factor) and bFGF (basic fibroblast growth factor). Fertilized White-Leghorn eggs were incubated under standardized conditions. Opening was performed at day three of incubation and preadipocytes with and without recombinant growth factors were transferred into a fibrin matrix and subsequently placed on the Chorioallantoic Membrane (CAM), respectively. Eight days later, the implanted constructs were explanted, histologically processed and vascularization evaluated by means of a computer-assisted image analysis program. Matrices containing preadipocytes displayed a significantly higher density of vascularization, whereas in the control group (fibrin without preadipocytes) no vessel ingrowth was observed. Daily application of recombinant growth factors added to the medium did not positively influence vascularization of the implant. Our investigations demonstrate that preadipocytes possess a strong angiogenic potential to induce and support neovascularization of 3D-fibrin matrices under in vivo conditions. Addition of recombinant growth factors did not result in any stimulatory effect. Neither did the application of fibrin alone demonstrate an angiogenic potential with regard to induction of vascularization.

The purpose of this study was to investigate the effect of platelet-rich-plasma (PRP) on cranial distraction osteogenesis. Standardized calvaria critical size defects (6 cm x 5 cm) were created in 16 adult female sheep. Bifocal cranial transport distraction osteogenesis with autogenous free calvaria bone grafts (2 cm x 4 cm) was performed at a rate of 1mm once daily to a total of 30 mm. The 16 sheep involved in the experiment were randomly divided into four groups, four animals in each: Group 1 (no PRP, latency 5 days); Group 2 (no PRP, latency 0 day), Group 3 (PRP, latency 5 days) and Group 4 (PRP, latency 0 day). After a consolidation phase of 6 weeks, the animals were sacrificed and specimens harvested for conventional radiological and 3D quantitative computer tomographic (3D-QCT) assessment. New bone was generated in the distraction zone in all groups. There were significantly (P < 0.05) higher densities in the proximal region of the distraction regenerate in Group 4 (PRP, latency 0 day) compared to Group 2. However, no significant differences in mean density of the total distraction regenerate were found, neither in volume of the bony regenerate between the experimental groups. This study showed that PRP only had an effect on bone regeneration if active distraction was started immediately after application of PRP in the distraction gap.

Introduction: Tissue engineering (TE) applications include the isolation, culture, and seeding of cells into a suitable matrix or scaffold prior to in vivo transplantation. After transplantation, vascularization of the scaffold is a principal factor limiting cell viability for the first 6-8 days post transplantation. A model has been developed for systematic analysis of this process., Methods: Fertilized white Leghorn eggs were incubated and opened at day 3 of incubation. Preadipocyte-seeded fibrin constructs were implanted in a specially designed plastic cylinder and placed through the opening onto the surface of the chorioallantoic membrane (CAM) at day 8 of incubation. Vascularization of the constructs by chorioallantoic blood vessels was assessed for up to 8 days post transplantation., Results: The survival rate for embryos receiving constructs was about 90%. Histology confirmed transplant cell viability at day 4 post transplantation, and vascularization of the constructs by avian endothelial cells progressively increased thereafter., Conclusion: A new in vivo model to study the effect of angiogenesis in TE constructs including assessments of viability, proliferation, and differentiation of transplanted cells and biomaterial properties is presented. Advantages include easy access to the CAM vascular network, lack of immunocompetence, low cost, and avoidance of animal experiments.

A volume-persistent culture of adipose tissue under in vivo conditions can be achieved only by early vascularization after cell transplantation. Cotransplantation of autologous preadipocytes with endothelial cells may enable the early formation of a capillary network. Investigations were performed in vivo in a specially adapted chorioallantoic membrane (CAM) model. Fertilized White Leghorn eggs were incubated and opened on day 3 of incubation and human dermal microvascular endothelial cell (HDMVEC) spheroids and preadipocytes were transferred in a fibrin matrix to the CAM. On day 7 after incubation the composites were explanted and immunohistologically investigated. Numerous vessels consisting of HDMVECs could be detected and the lumena of these vessels were perfused by chick erythrocytes. These results show the formation of a capillary network consisting of transplanted HDMVECs. The microcirculation of chick erythrocytes in vessels consisting of human endothelial cells proves the continuity of a newly formed capillary system to the host vessel system. The experiments demonstrate the first patent connection of tissue-engineered microvessels in adipose tissue to a host vessel system without applying exogenous angiogenic growth factors or transient transfection. The cotransplantation of endothelial cell spheroids with angiogenic mesenchymal cells may lead to the engineering of complex three-dimensional implants.

Tissue-engineering (TE) applications include the isolation, culture, and seeding of cells into a suitable matrix or scaffold before in vivo transplantation. After transplantation, vascularization of the scaffold is a principal limiting factor for cell viability for the first 6-8 days posttransplantation. A model for systematic analysis of this process has been developed. Fertilized White Leghorn eggs were incubated (at 37.8 degrees C in 60% relative humidity) and opened on day 3 of incubation. Preadipocyte-seeded fibrin constructs were implanted in a specially designed plastic cylinder and placed through the opening on the surface of the chorioallantoic membrane (CAM) on day 8 of incubation. Vascularization of the constructs by chorioallantoic blood vessels was assessed for up to 8 days posttransplantation. The survival rate for embryos receiving transplanted constructs was about 90%. Histology confirmed transplant cell viability at day 4 posttransplantation and vascularization of the constructs by avian endothelial cells began at this time. A new in vivo model to study the effect of angiogenesis in TE constructs, including assessments of viability, proliferation, and differentiation of transplanted cells and biomaterial properties, is presented. Advantages include easy access to the vascular network of the CAM, lack of immunocompetence, low costs, and avoidance of animal experiments.

Background: A review of a 10-year experience with the Ross procedure as a root replacement by a single group of surgeons featuring specific highlights is presented. Highlights include our results from a subset of patients with endocarditis and their management and a comparison of outcomes in patients with aortic insufficiency based on technical changes made after 5 years' experience., Methods: The total patient group was 191, with 148 male and 43 female participants with an age range from 1 day to 69 years. Five of the patients in the 0-to-20 age group were newborns. Fifty-three of the adults were operated on for infectious endocarditis. In the total patient group 43% had aortic insufficiency, 28% had aortic stenosis, and 29% had mixed disease., Results: Operative mortality was 5.2% with a late mortality of 2.6%. The actuarial survival was 90.2% at 10 years. Freedom from autograft explantation was 93.2% and freedom from homograft replacement was 98.4%. The endocarditis patients had an operative mortality of 3.8% with 100% cure of the infection. Freedom from reinfection on the autograft was 98.1%, and freedom from infection of the pulmonary homograft was 98.1%. The actuarial survival was 86.3%., Conclusions: A specific review of the patients with aortic insufficiency resulted in a failure of the autograft in 7 patients among a cohort of 41 during the first 5 years of the study. After a change in technique in which the aortic annulus is narrowed and fixed to a measured size to match the body surface area, we have had no failures in the autograft. Although these results are early, we believe that these data support the use of the autograft as an excellent choice for replacement of the aortic valve in infective endocarditis. Finally, the use of the autograft for aortic insufficiency is reasonable with fixation of the aortic annulus so that subsequent dilation does not occur.

Livers from hypophysectomized (hypox) rats were perfused with oxygenated Waymouth's medium in a system which permitted continuous recirculation for separate 30 minute periods after which fresh medium was supplied. In most experiments 6 changes of medium were carried out over a 3 hour period. The somatomedin activity of each perfusate was determined by measuring its ability to stimulate sulfate uptake in hypox rat cartilage in vitro. For comparison between experiments the results are expressed as the per cent stimulation of sulfate uptake by the perfusate compared with the unperfused buffer. Without hormonal additions there was a progressive fall in the release of somatomedin activity during the 6 periods of study. When compared with the results without hormone, the addition of 1000 muU/ml of insulin per ml of medium during the 2nd to 6th period led to a significant increase in perfusate somatomedin activity at all periods. The addition of 100 muU/ml of insulin was without significant effect. The possible inter-relationship between insulin and growth hormone in the regulation of somatomedin release was studied with a dose of bGH of 250 ng/ml which had previously been shown to be insufficient by itsel to stimulate somatomedin release. When added to a medium containing 1000 muU/ml of insulin, this dose of bGH did not significantly stimulate somatomedin release beyond that obtained with insulin alone. However, when 250 ng/ml was added to a medium containing 100 muU/ml insulin, a significant stimulation of somatomedin release was observed while the addition of each hormone separately was without significant effect. These results support the hypothesis that insulin shares with GH the regulation of somatomedin release by the liver. Differences in insulin concentration may explain some clinical situations in which somatomedin concentrations cannot be correlated with GH levels.