Tumor-penetrating peptide for systemic targeting of Tenascin-C.

Extracellular matrix in solid tumors has emerged as a specific, stable, and abundant target for affinity-guided delivery of anticancer drugs. Here we describe the homing peptide that interacts with the C-isoform of Tenascin-C (TNC-C) upregulated in malignant tissues. TNC-C binding PL3 peptide (amino acid sequence: AGRGRLVR) was identified by in vitro biopanning on recombinant TNC-C. Besides TNC-C, PL3 interacts via its C-end Rule (CendR) motif with cell-and tissue penetration receptor neuropilin-1 (NRP-1). Functionalization of iron oxide nanoworms (NWs) and metallic silver nanoparticles (AgNPs) with PL3 peptide increased tropism of systemic nanoparticles towards glioblastoma (GBM) and prostate carcinoma xenograft lesions in nude mice (eight and five-fold respectively). Treatment of glioma-bearing mice with proapoptotic PL3-guided NWs improved the survival of the mice, whereas treatment with untargeted particles had no effect. PL3-coated nanoparticles were found to accumulate in TNC-C and NRP-1-positive areas in clinical tumor samples, suggesting a translational relevance. The systemic tumor-targeting properties and binding of PL3-NPs to the clinical tumor sections, suggest that the PL3 peptide may have applications as a targeting moiety for the selective delivery of imaging and therapeutic agents to solid tumors.

Aneurysmal subarachnoid haemorrhage: effect of CRHR1 genotype on fatigue and depression.

BACKGROUND: Emotional health disturbances are common after aneurysmal subarachnoid hemorrhage (aSAH) and their causes are largely unexplored. Corticotropin-releasing hormone receptor 1 (CRHR1) is a key factor in stress reactivity and development of mental health disturbances after adverse life-events. METHODS: We explore the effect of CRHR1 genotype on mental health after aSAH in a retrospective cohort study. One hundred twenty-five patients have been assessed using EST-Q mental health questionnaire. Genotyping of CRHR1 single nucleotide polymorphisms (SNP-s) was performed (Rs7209436, Rs110402, Rs242924). RESULTS: Fatigue was present in almost half of aSAH patients, depression and anxiety in one-third. There was a high prevalence of insomnia and panic complaints. Rs110402 minor allele decreased the risk of depression (OR = 0.25, p = 0.027 for homozygotes). Depression was present in 14% vs 41% in minor and major allele homozygotes, respectively. Rs110402, Rs242924 and Rs7209436 minor alleles and TAT-haplotype, formed by them, were protective against fatigue. After Bonferroni correction only the association of Rs110402 with fatigue remained statistically significant (OR = 0.21, p = 0.006 for minor allele homozygotes). Results remained statistically significant when adjusted for gender, admission state, age and time from aSAH. In multiple regression analysis occurrence of fatigue was dependent on anxiety, modified Rankin score and Rs110402 genotype (R2 = 0.34, p <  0.001). CONCLUSIONS: CRHR1 minor genotype was associated with a lower risk of fatigue and depression after aSAH. Genetic predisposition to mental health disturbances associated with negative life-events could be a risk factor for fatigue and depression after aSAH and selected patients might benefit from advanced counselling in the recovery phase.

Aneurysmal subarachnoid haemorrhage: Effect of CRHR1 genotype on mental health-related quality of life.

Quality of life (QoL) disturbances are common after aneurysmal subarachnoid hemorrhage (aSAH) both in physical and mental health domains and their causes are not clearly understood. Corticotropin-releasing hormone receptor 1 (CRHR1) is involved in stress reactivity and development of mental health disturbances after negative life-events. We performed a retrospective cohort study of long-term QoL outcomes among 125 surgically treated aSAH patients (2001-2013). QoL was assessed with Short Form Health Survey (SF-36) and compared to an age and gender matched general population. Genotyping of CRHR1 single nucleotide polymorphisms was performed (Rs7209436, Rs110402, Rs242924) and their effect on QoL scores was explored. aSAH patients experienced a reduced quality of life in all domains. CRHR1 minor genotype was associated with higher SF-36 mental health (OR = 1.31-1.6, p < 0.05), role-emotional (OR = 1.57, p = 0.04) and vitality scores (OR = 1.31-1.38, p < 0.05). Association of all studied SNP's with vitality and Rs242924 with mental health scores remained statistically significant after Bonferroni correction. Mental quality of life scores were associated with physical state of patients, antidepressant history and CRHR1 genotype. Predisposition to mental health disturbances after stressful life-events might be associated with reduced mental QoL after aSAH and selected patients could be provided advanced counselling in the recovery phase.

Bi-specific tenascin-C and fibronectin targeted peptide for solid tumor delivery.

Oncofetal fibronectin (FN-EDB) and tenascin-C C domain (TNC-C) are nearly absent in extracellular matrix of normal adult tissues but upregulated in malignant tissues. Both FN-EDB and TNC-C are developed as targets of antibody-based therapies. Here we used peptide phage biopanning to identify a novel targeting peptide (PL1, sequence: PPRRGLIKLKTS) that interacts with both FN-EDB and TNC-C. Systemic PL1-functionalized model nanoscale payloads [iron oxide nanoworms (NWs) and metallic silver nanoparticles] homed to glioblastoma (GBM) and prostate carcinoma xenografts, and to non-malignant angiogenic neovessels induced by VEGF-overexpression. Antibody blockage experiments demonstrated that PL1 tumor homing involved interactions with both receptor proteins. Treatment of GBM mice with PL1-targeted model therapeutic nanocarrier (NWs loaded with a proapoptotic peptide) resulted in reduced tumor growth and increased survival, whereas treatment with untargeted particles had no effect. PL1 peptide may have applications as an affinity ligand for delivery of diagnostic and therapeutic compounds to microenvironment of solid tumors.

Peptide-guided nanoparticles for glioblastoma targeting.

Tumor-selective drug conjugates can potentially improve the prognosis for patients affected by glioblastoma (GBM) - the most common and malignant type of brain cancer with no effective cure. Here we evaluated a novel tumor penetrating peptide that targets cell surface p32, LinTT1 (AKRGARSTA), as a GBM targeting ligand for systemically-administered nanoparticles. LinTT1-functionalization increased tumor homing of iron oxide nanoworms (NWs) across a panel of five GBM models ranging from infiltratively-disseminating to angiogenic phenotypes. LinTT1-NWs homed to CD31-positive tumor blood vessels, including to transdifferentiated endothelial cells, and showed co-localization with tumor macrophages and lymphatic vessels. LinTT1 functionalization also resulted in increased GBM delivery of other types of systemically-administered nanoparticles: silver nanoparticles and albumin-paclitaxel nanoparticles. Finally, LinTT1-guided proapoptotic NWs exerted strong anti-glioma activity in two models of GBM, including doubling the lifespan of the mice in an aggressive orthotopic stem cell-like GBM that recapitulates the histological hallmarks of human GBM. Our study suggests that LinTT1 targeting strategy can be used to increase GBM uptake of systemic nanoparticles for improved imaging and therapy.

Influence of alarming auditory cues on viscoelastic stiffness of skeletal muscles in patients with Parkinson's disease.

BACKGROUND: Patients with Parkinson's disease can show brief normalization of motor activity in response to intense external stimuli - a phenomenon known as paradoxical kinesis. The purpose of the present study was to examine the effect of alarming auditory signals on the level of viscoelastic stiffness of skeletal muscles as an indicator of parkinsonian rigidity. METHODS: Myotonometry was used to determine the changes of viscoelastic stiffness of skeletal muscles in ten patients in an advanced stage of Parkinson's disease, treated with deep brain stimulation, and ten healthy controls. The measurements were repeated and compared during the stimulation-on and stimulation-off periods, with and without auditory alarming signals. FINDINGS: The mean values of stiffness measured in the stimulation-off phase (370.4 N/m) were significantly higher than the values obtained in the stimulation-on phase (339.2 N/m) (q = 6.05; P < 0.01) but also in the stimulation-off with alarming signals phase (349.6 N/m) (q = 4.04; P < 0.05). In the normal controls, exposure to the auditory alarming signals did not change the values of viscoelastic stiffness. INTERPRETATION: These findings demonstrate that the phenomenon of paradoxical kinesis is associated with the changes of muscular rigidity in parkinsonian patients. Results from the study may help to establish new strategies for addressing motor disabilities in patients with Parkinson's disease.

The effect of subthalamic stimulation on viscoelastic stiffness of skeletal muscles in patients with Parkinson's disease.

BACKGROUND: Myotonometric evaluation of viscoelastic stiffness of skeletal muscles has been proposed to document the effect of surgical or pharmacological treatment on rigidity in patients with Parkinson's disease. The aim of the study was to analyze the changes of viscoelastic stiffness induced by deep brain stimulation. METHODS: Fifteen patients in an advanced stage of Parkinson's disease participated in the study. The study took place in the off-medication conditions after one night of drug withdrawal. The Unified Parkinson's Disease Rating Scale was used for clinical assessment of the disease. Myotonometry was used to measure viscoelastic stiffness in the resting muscles before and directly after passive wrist movements, commonly used for clinical evaluation of rigidity. The measurements were repeated during the stimulation-on and stimulation-off periods and compared with fifteen healthy control persons. FINDINGS: The clinical scores for wrist rigidity improved from 3.0 (1-4) to 0.93 (0-2) (P<0.05) due to brain stimulation. The mean values of viscoelastic stiffness were similar before and after passive wrist movements, but the differences between the patients with high vs. low rigidity values (354.9 vs 310.2N/m; P<0.05) and in stimulation-off vs. stimulation-on conditions (342.7 vs 310.5N/m; P<0.05) were significant only if the measurements had been performed after passive wrist movements. INTERPRETATION: Effective deep brain stimulation and increased rigidity can significantly change viscoelastic stiffness in the resting muscles in patients with Parkinson's disease, especially if evaluated after passive wrist movements. This paper supports the use of myotonometry for objective quantification of parkinsonian rigidity at rest.

Placebo effects induced by auditory cues decrease parkinsonian rigidity in patients with subthalamic stimulation.

Placebo effects are the consequence of an interaction between an organism and its surroundings and may be influenced by cues from the environment. Our study was designed to analyze if conditioned auditory cues could trigger placebo effects and affect parkinsonian rigidity as measured by viscoelastic properties of skeletal muscles in patients treated with subthalamic stimulation. We found that after repeatedly associating with the effect of deep brain stimulation on rigidity, a common dial phone signal itself was able to reduce the mean values of viscoelastic stiffness in the placebo stage (368.8±50.4Nm(-1)) as compared to the stimulation-off conditions (383.7±61.2Nm(-1)) (q=4.18; p<0.05) in ten patients with Parkinson's disease. Thus, it appears that due to associative learning processes environmental cues can acquire the capacity to trigger placebo effects affecting the clinical status of the patients.

Patient involvement in surgical treatment decisions and satisfaction with the treatment results after lumbar intervertebral discectomy.

PURPOSE: To evaluate involvement of patients in surgical treatment decision making (STDM) in relations to satisfaction with the results of lumbar discectomy. METHODS: We evaluated prospectively 150 surgically treated patients with radicular pain due to lumbar disc herniation (LDH). The patients completed self-reported questionnaires about preferences for involvement and actual involvement in STDM. Global satisfaction with the results of the treatment was assessed at 1 year after the operation. RESULTS: Most of the patients (129 patients, 86%) stated that they had been sufficiently informed about LDH to be involved in the treatment decisions, almost half of the patients (47%) preferred active or collaborative involvement and 58% of the patients reported higher actual involvement in STDM. Congruence between preferred and actual roles in decision making was 64%. Most of the patients (77%) were satisfied with the results of the operation, but satisfaction was not associated with involvement of patients in STDM. CONCLUSION: A significant proportion of patients with LDH prefer to be actively involved in treatment decisions and experience an STDM process that matches their preferences for participation. However, individual differences in preferences for involvement in STDM are common and global satisfaction with the treatment results is not significantly related to the activity of involvement in STDM.

Relationship between neovascularization and degenerative changes in herniated lumbar intervertebral discs.

PURPOSE: Lumbar disc degeneration may be associated with intensity of neovascularization in disc herniations. Our study was designed to evaluate how much the severity of histodegeneration is related to the development of neovascularization and to the level of pleiotrophin in the herniated lumbar discs. METHODS: Surgically excised lumbar disc specimens were obtained from 29 patients with noncontained (i.e., extruding through the posterior longitudinal ligament) and 21 patients with contained disc herniations. The histodegeneration scores and levels of neovascularization were estimated according to semiquantitative analysis in lumbar disc and endplate samples. Immunohistochemical staining were performed to identify the newly formed blood vessels and to detect the presence of pleiotrophin in the specimens. RESULTS: Higher levels of disc and endplate neovascularity were registered in noncontained herniations. The level of neovascularization was significantly related to the score of histodegeneration in the herniated disc tissues but not in the endplate specimens. Both contained and noncontained herniations had the highest values of histodegeneration in conjunction with the highest level of neovascularization but the relations between neovascularity and degenerative changes remained to be significant only in the group of noncontained herniations. Registration or frequency of pleiotrophin positive cells did not correlate significantly with histodegeneration or level of neovascularization in the disc samples. CONCLUSION: Severe histodegeneration of the lumbar disc herniations is associated with enhanced neovascularization and potentially also spontaneous regression of the herniated tissue.

Emotional health and quality of life after aneurysmal subarachnoid hemorrhage.

BACKGROUND: Emotional disorders and decrease in health-related quality of life (HRQoL) are well-documented sequelae of aneurysmal subarachnoid hemorrhage (SAH). The aim of this study was to analyze the impact of emotional disorders on HRQoL in survivors of SAH. METHODS: This was a retrospective study enrolling 114 SAH-survivors at 1-10 years (mean 4,5 years) after the disease. Emotional State Questionnaire (EST-Q) was used to measure emotional health and the Short Form Health Survey (SF-36) was used to assess HRQoL of the patients. RESULTS: Most of the patients reported good recovery, but EST-Q results revealed high prevalence of emotional disorders after SAH. Almost half of the patients had higher than cut-off values indicating fatigue (47 %) and insomnia (46 %). About one third of SAH-patients had higher than cut-off scores demonstrating depression (30 %) and anxiety (31 %). The patients scored significantly lower in all scales of SF-36 as compared to age-matched general population. All EST-Q subscale results were significantly and negatively correlated with SF-36 scores. Fatigue was independently related to all SF-36 subscales and depression to most of the mental health component scores. Emotional symptoms alone were demonstrated to explain 23-47 % of the SF-36 subscale values, and more than half of the variance of mental health component score values were found to be explained by emotional disorders (53 %). CONCLUSIONS: Emotional disorders are frequent after SAH and significantly associated with impairment of HRQOL. Proper and timely screening tests are important to reveal development of emotional problems and improve QoL for the SAH-patients.

Postoperative spondylodiscitis caused by Mycobacterium bovis BCG: a case study.

BACKGROUND CONTEXT: Postoperative spondylodiscitis (PSD) is a known complication of lumbar disc surgery. The etiology of the disease is usually bacterial, but several uncommon infectious agents have also been described; however, there are no reports about postoperative colonization with Mycobacterium bovis bacille Calmette-Guérin after lumbar discectomy. PURPOSE: To describe the case of PSD caused by M. bovis BCG, and to discuss diagnostic and therapeutic interventions as well as possible pathogenic mechanisms of the disease. STUDY DESIGN: Case report and review of the literature. METHODS: A 31-year-old man was operated on because of L4-L5 lumbar disc herniation. Two months later, the patient presented with gradual increase of back pain, and magnetic resonance imaging confirmed PSD. He started to receive antibacterial treatment and was reoperated on because of progressive neurological deficits due to epidural abscess. Neurological status improved, but a fistula developed with intermittent pus drainage from the operative scar. RESULTS: Microbiological cultures were repeatedly obtained from the pus, but all the initial stains and cultures were negative. Four months after the reoperation, the culture for M. bovis BCG from the pus appeared to be positive. The patient received antituberculosis regimen, including isoniazid, rifampin, ethambutol, and ofloxacin. The clinical symptoms resolved, and antituberculosis treatment was discontinued after 14 months. CONCLUSIONS: Mycobacterium bovis BCG must be considered in the differential diagnosis of PSD. Microbiological analysis and radiological studies are vital components in diagnosis; if there is any suspicion of BCG osteomyelitis, proper diagnostic and therapeutic management must be instituted without delay to avoid an unfavorable outcome.

Subthalamic stimulation improves the cerebral hemodynamic response to the cold pressure test in patients with Parkinson's disease.

PURPOSE: Disturbances of the autonomic nervous system are common in patients with Parkinson's disease (PD) but the effect of deep brain stimulation of the subthalamic nucleus on cerebrovascular reactivity is not entirely known. METHODS: Seven patients in an advanced stage of the disease and seven healthy age-matched controls participated in the study, which took place after one night of drug withdrawal. Cerebral blood flow velocity was continuously monitored on both sides with transcranial Doppler ultrasound, and cerebrovascular reactivity (CR) was evaluated with the cold pressure test. The measurements were repeated and compared during the stimulation-on and -off phases. RESULTS: The PD patients had significantly higher CR values in the stimulation-on than -off conditions (15.1% ± 6.9 versus 9.4% ± 6.2; p = 0.03). CR values were higher in controls than in patients in the stimulation-off condition (20.4% ± 12.5 versus 9.4% ± 6.2; p = 0.007) without a significant difference with the stimulation-on phase. CONCLUSIONS: CR, evaluated by the response to the cold pressure test, is impaired in patients with advanced PD and improved by subthalamic nucleus.

Changes in viscoelastic properties of skeletal muscles induced by subthalamic stimulation in patients with Parkinson's disease.

BACKGROUND: Objective measurements would be useful to document the effect of deep brain stimulation in alleviating rigidity in patients with Parkinson's disease. The aim of the study was to examine the changes of viscoelastic properties in skeletal muscles as indicators of rigidity. METHODS: Six patients in an advanced stage of Parkinson's disease participated in the study. The study took place in the off-medication conditions after one night of drug withdrawal. The wrist rigidity was examined according to the Unified Parkinson's Disease Rating Scale in both sides. Myotonometry (Myoton) was used to determine stiffness and elasticity in extensor digitorum muscles bilaterally. The measurements were repeated and compared during the stimulation-on and stimulation-off periods. FINDINGS: A comparison of mean clinical motor scores revealed a significant improvement of parkinsonian symptoms due to brain stimulation. In particular, arm rigidity improved on average from 2.83 (1-4) in stimulation-off phase to 1.17 (0-2) in stimulation-on phase (P<0.05). The mean values of elasticity and stiffness were not significantly different in stimulation-on and stimulation-off conditions. The patients with elevated clinical rigidity scores had higher mean values of stiffness (262.5 vs 211.0; P<0.05) but the differences in elasticity were not significant. INTERPRETATION: Increased rigidity is associated with increased values of viscoelastic stiffness. This paper supports the use of myotonometry for objective quantification of rigidity and in the future, this tool could prove helpful for optimizing deep brain stimulation settings in patients with Parkinson's disease.