The Neuroblastoma Microenvironment, Heterogeneity and Immunotherapeutic Approaches.

Neuroblastoma is a peripheral nervous system tumor that almost exclusively occurs in young children. Although intensified treatment modalities have led to increased patient survival, the prognosis for patients with high-risk disease is still around 50%, signifying neuroblastoma as a leading cause of cancer-related deaths in children. Neuroblastoma is an embryonal tumor and is shaped by its origin from cells within the neural crest. Hence, neuroblastoma usually presents with a low mutational burden and is, in the majority of cases, driven by epigenetically deregulated transcription networks. The recent development of Omic techniques has given us detailed knowledge of neuroblastoma evolution, heterogeneity, and plasticity, as well as intra- and intercellular molecular communication networks within the neuroblastoma microenvironment. Here, we discuss the potential of these recent discoveries with emphasis on new treatment modalities, including immunotherapies which hold promise for better future treatment regimens.

Cognitive function in amyotrophic lateral sclerosis: a cross-sectional and prospective pragmatic clinical study with review of the literature.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) can present with either bulbar or spinal symptoms, and in some cases, both types of symptoms may be present. In addition, cognitive impairment has been observed in ALS. The study aimed to evaluate the frontal and general cognitive performance in ALS not only cross-sectionally but also longitudinally. METHODS AND MATERIALS: The Frontal Assessment Battery (FAB) and the Montreal Cognitive Assessment (MoCA) were employed to assess cognitive function in 52 adults with ALS and 52 cognitively healthy individuals. The statistical analyses encompassed the Pearson Chi square test, the Skillings-Mack test, the Spearman's rank correlation coefficient, and the Proportional Odds Logistic Regression Model (POLR). RESULTS: Cross-sectionally, lower cognitive performance was associated with ALS diagnosis, older age, and motor functional decline. The cognitive impairment of individuals with bulbar and spinal-bulbar symptoms showed faster deterioration compared to those with spinal symptoms. The spinal subgroup consistently performed worst in delayed recall and attention, while the spinal-bulbar and bulbar subgroups exhibited inferior scores in delayed recall, attention, visuospatial skills, orientation, and verbal fluency. CONCLUSION: The incorporation of cognitive screening in the diagnostic workup of ALS may be beneficial, as early detection can enhance symptom management and improve the quality of life for both individuals with ALS and their care partners.

The role of cerebrospinal fluid levels of neutrophil gelatinase-associated lipocalin (NGAL) and electroencephalography in the assessment of impaired consciousness in the context of infection.

INTRODUCTION: The sepsis syndrome is potentially affecting several organs and systems irrespectively of the primary source of the infection. Alterations of the brain function in sepsis patients may result either from a primary central nervous system (CNS) infection or could be part of the sepsis-associated encephalopathy (SAE), a common complication of sepsis, characterized by a diffuse dysfunction of the brain due to an infection elsewhere in the body without overt CNS infection. Aim of the study was to evaluate the usefulness of electroencephalography and the biomarker neutrophil gelatinase-associated lipocalin (NGAL) when measured in the cerebrospinal fluid (CSF) in the management of these patients. METHODS: Patients presenting at the emergency department with altered mental status and signs of infection were included in this study. Among initial assessment and treatment of the patients based on the international guidelines for treating sepsis, NGAL was measured in the cerebrospinal fluid (CSF) using ELISA technique. Electroencephalography was performed when possible within 24 hours after admission and EEG abnormalities were recorded. RESULTS: 32 of 64 patients included in this study were diagnosed with central nervous system (CNS) infection. CSF NGAL was significantly higher in patients with CNS infection compared to patients without CNS infection (18.1 [5.1-71.1] vs 3.6 [1.2-11.6]; p<0.001). There was a trend for higher CSF NGAL in patients with EEG abnormalities, which did not reach statistical significance (p=0.106). CSF NGAL levels were similar between survivors and non-survivors (medians: 7.04 vs 11.79). CONCLUSION: In patients presenting at the emergency department with altered mental status and signs of infection, CSF NGAL was significantly higher in patients with CSF infection. Its role in this acute setting should be evaluated further. CSF NGAL could be suggestive of EEG abnormalities.

Α Multicenter Retrospective Study Evaluating Brivaracetam in the Treatment of Epilepsies in Clinical Practice.

Brivaracetam (BRV) is the latest approved antiepileptic drug. The aim of the study was to evaluate the efficacy and tolerability of BRV in everyday clinical practice. In this retrospective, observational, multicenter study, data from epilepsy patients receiving BRV from January 2018 to July 2019 were analyzed. Patients with age ≥16 suffering from any type of epilepsy and having at least one follow up encounter after dose titration were included. 156 consecutive patients were included in the study. The mean age was 40 (16-84 years) and the mean duration of epilepsy was 21 years. Of the 156 patients, 81% were diagnosed with focal-onset seizures, 16% with generalized seizures, while 3% suffered from unclassified seizures. Nine patients received BRV as monotherapy as a switching therapy. At the first follow up visit, seizure cessation was achieved in 56 (36%) patients and the rate of ≥50% responders was 36%. Twenty four patients (15%) remained unchanged; six patients (4%) were recorded with increased seizure frequency, while the remaining 9% had a response of less than 50%. Twenty-six patients (17%) showed clinically significant adverse events, but none were life threatening. Brivaracetam seems to be an effective, easy to use and safe antiepileptic drug in the clinical setting.

The optimum hand temperature to study nerve conduction in patients with carpal tunnel syndrome.

To define the skin temperature at which diseased nerves are better differentiated from the healthy. Motor and sensory conduction of median and ulnar nerve were evaluated in 52 patients with carpal tunnel syndrome (CTS) and 52 matched healthy controls at environmental skin temperature (mean 32-33 °C), after warming by an average of 2 °C and cooling to approximately 6 °C below baseline. In the hot condition, group comparisons for the median nerve showed a similar rate of distal motor latency (DML) reduction and sensory conduction velocity (SCV) increase in CTS and controls. With cold, the rate of change was smaller for the patients: DML mean increase was 5% /°C (7% for controls) and SCV mean decrease was 2.5%/°C (3.2% for controls). Individual patients' analysis revealed fewer abnormal median DML and SCV values at hot or at cold, compared to environmental temperature. It is concluded that conduction adjustments for low hand temperatures based on healthy measurements resulted in overcorrection and therefore underdiagnosis of CTS. Alternatively, at excessive hand warming the convergence of patient and healthy measurements also lead to underdiagnosis. Maintenance of skin temperature at 32-33 °C, corresponding to normal body temperature, is the optimum approach and should always be employed in clinical practice.

Efficient identification of Acetylcholinesterase and Hyaluronidase inhibitors from Paeonia parnassica extracts through a HeteroCovariance Approach.

ETHNOPHARMACOLOGICAL RELEVANCE: On the basis of the relevant reference in the poem Theriaca of the ancient Greek physician Nicander and its traditional use, Paeonia parnassica was selected for the evaluation of two extracts obtained from the roots and aerial parts to inhibit hydrolytic enzymes involved in snake envenomation. The secondary metabolites which contribute to these activities were detected through a novel HeteroCovariance NMR based approach. Afterwards these ingredients were isolated, identified and evaluated for their inhibitory potency. AIM OF THE STUDY: The identification of acetylcholinesterase and hyaluronidase inhibitors from Paeonia parnassica extracts was used as a case study for the introduction of a recently developed methodology to evaluate ethnopharmacological data and exploit them for the discovery of bioactive natural compounds. This process is based on the fractionation of the selected extracts and the simultaneous phytochemical analysis and biological assessment of the resulting fractions, which permits the rapid detection of the specified secondary metabolites prior to any laborious and time-consuming purification. MATERIALS AND METHODS: The roots and aerial parts of P. parnassica were extracted using methanol: water 50:50 and the two resulted extracts were fractionated by Centrifugal Partition Chromatography. The obtained fractions were evaluated in-vitro for their ability to inhibit acetylcholinesterase and hyaluronidase enzymes and their 1H NMR spectra were recorded. The biological activity was statistically correlated with the spectral data through the HeteroCovariance Approach (HetCA). Finally the purification, identification and biological evaluation of targeted secondary metabolites were carried out. RESULTS: The general chemical structures and some explicit secondary metabolites which contribute (e.g. gallotannins, gallic acid derivatives) or not (characteristic "cage-like" monoterpenes of the genus, glycosylated flavonoids) to the anti-acetylcholinesterase and anti-hyaluronidase activities were detected through HetCA. The consequent isolation and biological evaluation of targeted compounds were performed in order to validate the effectiveness and precision of the methodology. This procedure revealed the most active ingredients of both extracts obtained from roots and aerial parts against the above mentioned biological targets, as well as other compounds possessing moderate activity. CONCLUSIONS: The results of this study contributed to the verification of the ancient text Theriaca regarding the use of Paeonia parnassica to treat the snake bite symptoms. Furthermore, the ingredients of the Paeonia parnassica extracts, which were responsible for their anti-cholinesterase and anti-hyaluronidase activities, were determined applying a HetCA methodology before their isolation. Therefore, the current work provides clear evidence that HetCA could consist an efficient tool for the exploitation of traditional medicine information in order to discover bioactive natural compounds and develop new pharmacotherapies which serve the needs of contemporary medicine.

Selective cytotoxicity of the herbal substance acteoside against tumor cells and its mechanistic insights.

Natural products are characterized by extreme structural diversity and thus they offer a unique source for the identification of novel anti-tumor agents. Herein, we report that the herbal substance acteoside being isolated by advanced phytochemical methods from Lippia citriodora leaves showed enhanced cytotoxicity against metastatic tumor cells; acted in synergy with various cytotoxic agents and it sensitized chemoresistant cancer cells. Acteoside was not toxic in physiological cellular contexts, while it increased oxidative load, affected the activity of proteostatic modules and suppressed matrix metalloproteinases in tumor cell lines. Intraperitoneal or oral (via drinking water) administration of acteoside in a melanoma mouse model upregulated antioxidant responses in the tumors; yet, only intraperitoneal delivery suppressed tumor growth and induced anti-tumor-reactive immune responses. Mass-spectrometry identification/quantitation analyses revealed that intraperitoneal delivery of acteoside resulted in significantly higher, vs. oral administration, concentration of the compound in the plasma and tumors of treated mice, suggesting that its in vivo anti-tumor effect depends on the route of administration and the achieved concentration in the tumor. Finally, molecular modeling studies and enzymatic activity assays showed that acteoside inhibits protein kinase C. Conclusively, acteoside holds promise as a chemical scaffold for the development of novel anti-tumor agents.

Electroencephalographic Abnormalities in Sepsis Patients in Correlation to the Calculated Prognostic Scores: A Case Series.

OBJECTIVE: To evaluate the electroencephalographic (EEG) findings and correlate EEG findings with inflammatory biomarkers and the sepsis prognostic scores SOFA, SAPS II and APACHE II in patients who present in the Emergency Department with sepsis without clinical central nervous system involvement. METHODS: The study included seventeen patients (< 70 years old) with sepsis without central nervous system involvement presenting in the Emergency Department of the University Hospital of Patras, Greece. All patients underwent neurologic examination and EEG analysis on admission to the hospital and were treated according to the international guideline protocols for sepsis. RESULTS: Six of seventeen sepsis patients had mild or moderate EEG abnormalities. We did not find any significant correlation between EEG abnormalities and inflammatory biomarkers (CRP, WBC) or commonly used prognostic sepsis scores. CONCLUSIONS: EEG could serve as a useful tool to identify brain alterations at an early stage in sepsis, before clinical sings of encephalopathy can be detected. However, the presence of EEG abnormalities does not correlate with sepsis severity as measured by the commonly used prognostic sepsis scores SOFA, APACHE II or SAPS II. Because this was a small single center observational study, large multi-center studies are warranted to confirm these findings.

Development of a Sustainable Procedure for the Recovery of Hydroxytyrosol from Table Olive Processing Wastewater Using Adsorption Resin Technology and Centrifugal Partition Chromatography.

The present endeavor aims to establish a novel procedure, applicable to the extraction and isolation of hydroxytyrosol from table olive processing wastewater. A two-step chromatographic separation is presented using non-ionic absorbent resin for the recovery of its phenolic content, followed by purification of hydroxytyrosol with centrifugal partition chromatography. Two table olive processing wastewaters, obtained from Kalamon and Amfissis olive varieties, were used. In the extracts obtained after resin treatment, the hydroxytyrosol content was determined by high-performance liquid chromatography with diode-array detection to be 4.05% and 10.10%, respectively. The extract from Amfissis table olive processing wastewater was further processed with preparative centrifugal partition chromatography for the purification of hydroxytyrosol. High-performance liquid chromatography analysis revealed that the isolated compound was >95% purity.

Treatment of Tardive Dystonia Induced by Antipsychotics, Old and New.

OBJECTIVES: Tardive dystonia is a serious extrapyramidal side effect emerging after long-term treatment with antipsychotics, frequently with a deteriorating course, and unsatisfactory treatment. Presently, clozapine is used for the cotreatment of tardive dystonia and psychosis, at the cost of serious side effects. Apart from clozapine, there have been case reports describing positive effects of quetiapine on dystonic symptoms. Aim of the present study was to demonstrate the ameliorating effects of quetiapine on dystonic symptoms, in a sample of patients suffering from antipsychotic-induced tardive dystonia. METHODS: Quetiapine was administered to 16 consecutively enrolled stabilized patients with psychotic or mood disorders and tardive dystonia, replacing the "offending drugs," over a 3-month cross-tapering period. Target dose of quetiapine was set according to the defined daily dose of the received antipsychotic(s) at baseline, as reviewed by the World Health Organization Center of Drug Statistics Methodology, aiming at both maintenance of psychosis control and reduction of dystonic symptoms. RESULTS: Patients were found to have significant positive results in amelioration of dystonia (P < 0.001) over a 1-year period, without loss of antipsychotic efficacy. Reduction of dystonic symptoms with the use of quetiapine could be considered comparable with the positive effects of clozapine, with the additional advantage of relatively lacking serious side effects. CONCLUSIONS: Quetiapine may represent a valuable therapeutic choice for the treatment of tardive dystonia.

Pure sensory chronic inflammatory polyneuropathy: rapid deterioration after steroid treatment.

BACKGROUND: Chronic inflammatory demyelinating polyneuropathy (CIDP) as a pure sensory variant is rarely encountered. Therefore the best treatment option is hard to define. CASE PRESENTATIONS: We reported two middle-aged patients of Caucasian origin, one female and one male, who over a period of several months presented limbs and gait ataxia. Clinical and neurophysiological examination revealed only sensory abnormalities. A diagnosis of atypical CIDP was suggested, considering the elevated CSF protein level and the presence of anti-gangliosides antibodies. Ten and 15 days respectively after initiation of prednisolone treatment both patients experienced exacerbation of sensory symptoms and emerging of muscle weakness. Steroids were then substituted by rituximab in the first patient and intravenous immunoglobulin in the second patient resulting in gradual decrement of symptoms and signs. Two-year follow-up showed no further deterioration. CONCLUSION: Caution should be exercised when treating cases of pure sensory polyneuropathy with high dose steroids since an unfavorable outcome is possible.

Herbal substance, acteoside, alleviates intestinal mucositis in mice.

This study investigated the role of acteoside in the amelioration of mucositis. C57BL/6 mice were gavaged daily with acteoside 600 µg for 5 d prior to induction of mucositis and throughout the experimental period. Mucositis was induced by methotrexate (MTX; 12.5 mg/kg; s.c.). Mice were culled on d 5 and d 11 after MTX. The duodenum, jejunum, and ileum were collected for myeloperoxidase (MPO) activity, metallothionein (MT) levels, and histology. Acteoside reduced histological severity scores by 75, 78, and 88% in the duodenum, jejunum, and ileum, respectively, compared to MTX-controls on d 5. Acteoside reduced crypt depth by 49, 51, and 33% and increased villus height by 19, 38, and 10% in the duodenum, jejunum, and ileum, respectively, compared to MTX-controls on d 5. Acteoside decreased MT by 50% compared to MTX-control mice on d 5. Acteoside decreased MPO by 60% and 30% in the duodenum and jejunum, respectively, compared to MTX-controls on d 5. Acteoside alleviated MTX-induced small intestinal mucositis possibly by preventing inflammation.

Leishmanicidal activity assessment of olive tree extracts.

Leishmaniasis, a protozoan parasitic disease that remains a major worldwide health problem with high endemicity in developing countries, is prevalent around the Mediterranean basin. High cost, systemic toxicity, and diminished efficacy due to development of parasite resistance are the serious drawbacks of current treatment options. Thus, identifying new, effective, and safer anti-leishmanial drug(s) is of paramount importance. Here we tested the anti-promastigote and anti-amastigote activity of five natural products, including oleuropein and hydroxytyrosol, present in olive tree leaves and olive mill wastewater. These products are recognized as low-cost starting materials rich in bioactive compounds, particularly biophenols. Oleuropein and hydroxytyrosol exhibited the best inhibitory effect among the natural products tested in both stationary and middle logarithmic phase promastigotes of L. infantum, L. donovani, and L. major. Similarly, oleuropein and hydroxytyrosol demonstrated the highest selectivity index ratio against L. donovani amastigotes that parasitize J774A.1 macrophages. Moreover, oleuropein was tested in vivo in an experimental visceral leishmaniasis model. L. donovani-infected BALB/c mice received intraperitoneal oleuropein a total of 14 times at intervals of every other day. Three days after treatment termination, the spleen parasitic burden was reduced >80%. Of interest, this effect of oleuropein persisted and was even enhanced 6 weeks after the termination of the treatment, as determined by parasite depletion of >95% in liver and spleen. These findings contribute to the potential development of natural products as effective drugs against parasites of the Leishmania genus, with low cost and diminished cytotoxicity.

Frontotemporal dementia, manifested as schizophrenia, with decreased heterochromatin on chromosome 1.

Introduction. Frontotemporal dementia is a disorder of complex etiology, with genetic components contributing to the disease. The aim of this report is to describe a young patient suffering from frontotemporal dementia, misdiagnosed as schizophrenia, related to a genetic defect on chromosome 1. Case Presentation. A 29-year-old female patient, previously diagnosed as having schizophrenia, was hospitalized with severe behavioural disturbances. She demonstrated severe sexual disinhibition, hyperphagia, lack of motivation, apathy, psychotic symptoms, suicidal thoughts, and cognitive deterioration. Focal atrophy of frontal and anterior temporal structures bilaterally was found on brain MRI, as well as bifrontal hypo perfusion of the brain on SPECT scan. The diagnosis of frontotemporal dementia was made clinically, according to Lund and Manchester groups and Neary diagnostic criteria. Chromosomal analysis was conducted and revealed decrease in length of heterochromatin on the long arm of chromosome 1 (46, XX, 1qh-). Parental karyotypes were normal. Discussion. Frontotemporal dementia, and particularly early-onset cases, can be often misdiagnosed as schizophrenia, with negative impact on case management. Genetic testing could be an aid to the correct diagnosis, which is crucial for optimal patient care.

Indirubins deplete striatal monoamines in the Intact and MPTP-treated mouse brain and block kainate-induced striatal astrogliosis.

The indirubins long have been used in Chinese medicine for treatment of myelocytic leukemia. Among the many more recently described biological activities of the indirubins, attention has been directed toward the ability of these compounds to inhibit GSK-3 and CDKs, kinases implicated in neurodegenerative conditions. Little information is available on effects of indirubins on chemically-induced neurodegeneration. Here we examined the influence of three indirubins on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)- and kainic acid (KA)-induced neurotoxicity in the mouse. The three indirubins examined were 6-bromoindirubin-3'-oxime (6BIO), 5-bromoindirubin-3'-oxime (5BIO) and 5-amino-6-bromoindirubin (5A6BI). The first two derivatives were previously described indirubins with low nanomolar inhibitory activity against GSK-3 and CDKs. The third compound was synthesized by the dimerization of 5-amino-6-bromoisatin with 3-acetoxyindol. The synthesis of the key compound 5-amino-6-bromoisatin was based on the bromination of the ketal of 5-amino-isatin. All indirubins examined decreased various measures associated with dopaminergic neurotransmission in striatum. These effects occurred alone or over and above the decrements seen following administration of the dopaminergic neurotoxicant, MPTP. Striatal serotonin and serotonin turnover were decreased by the indirubins in MPTP-treated mice. None of these striatal effects of the indirubins alone were associated with evidence of astrogliosis, an indicator of underlying neuropathology, nor did they potentiate the astrogliosis accompanying administration of MPTP. In general, the indirubins reduced KA-associated mortality and striatal but not hippocampal astrogliosis due to this toxicant. The data suggest that indirubins affect striatal biogenic amine levels and turnover in intact mice. The data do not indicate a neuroprotective action of indirubins in mice treated with MPTP but that they do suggest that they may be neuroprotective against KA-induced injury of the neostriatum.

Chronotypology and melatonin alterations in minimal hepatic encephalopathy.

BACKGROUND: "Minimal (subclinical) hepatic encephalopathy" is a term that describes impairment of every day life activities in cirrhosis patients without clinical neurologic abnormalities. Melatonin diurnal pattern disruption and metabolic changes due to liver insufficiency can affect the human biologic clock. Our study was conducted to measure plasma melatonin levels in an attempt to correlate plasma melatonin abnormalities with liver insufficiency severity, and describe chronotypology in cirrhosis patients with minimal encephalopathy. METHODS: Twenty-six cirrhotic patients enrolled in the study and thirteen patients without liver or central nervous system disease served as controls. All patients had full clinical and biochemical evaluation, chronotypology analysis, neurological evaluation, melatonin profile and quality of life assessment. RESULTS: Cirrhotic patients with minimal encephalopathy exhibit melatonin secretion abnormalities. Cirrhosis patients with more severe hepatic insufficiency (Child-Pugh score > 5) had significantly (p < 0.04) lower evening melatonin levels compared to patients with less severe insufficiency (Child-Pugh score = 5).Chronotypology analysis revealed Morning Type pattern in 88% of cirrhosis patients. DISCUSSION: The presence of abnormal plasma melatonin levels before the onset of clinical hepatic encephalopathy, and the finding that patients with more severe cirrhosis have lower evening melatonin levels are the most important findings of this study. Despite these melatonin abnormalities, chronotypology revealed Morning Type pattern in 23 of 26 cirrhosis patients. We believe these findings are important and deserve further study. CONCLUSION: Melatonin abnormalities occur in cirrhosis patients without clinical encephalopathy, are related to liver insufficiency severity, may influence chronotypology patterns, and certainly deserve further investigation.

Pituitary hormone circadian rhythm alterations in cirrhosis patients with subclinical hepatic encephalopathy.

AIM: To analyze pituitary hormone and melatonin circadian rhythms, and to correlate hormonal alterations with clinical performance, hepatic disease severity and diagnostic tests used for the detection of hepatic encephalopathy in cirrhosis. METHODS: Twenty-six patients with cirrhosis were enrolled in the study. Thirteen patients hospitalized for systemic diseases not affecting the liver were included as controls. Liver disease severity was assessed by the Child-Pugh score. All patients underwent detailed neurological assessment, electroencephalogram (EEG), brain magnetic resonance imaging (MRI), assays of pituitary hormone, cortisol and melatonin, and complete blood chemistry evaluation. RESULTS: Pituitary hormone and melatonin circadian patterns were altered in cirrhosis patients without clinical encephalopathy. Circadian hormone alterations were different in cirrhosis patients compared with controls. Although cortisol secretion was not altered in any patient with cirrhosis, the basal cortisol levels were low and correlated with EEG and brain MRI abnormalities. Melatonin was the only hormone associated with the severity of liver insufficiency. CONCLUSION: Abnormal pituitary hormone and melatonin circadian patterns are present in cirrhosis before the development of hepatic encephalopathy. These abnormalities may be early indicators of impending hepatic encephalopathy. Factors affecting the human biologic clock at the early stages of liver insufficiency require further study.

Persistent suppression of postexercise facilitation of motor evoked potentials during alternate phases of bipolar disorder.

The current study sought to longitudinally evaluate the postexercise facilitation of motor evoked potentials (MEP) in two patients during different phases of short-circle depressive-manic disorder. Each study included 50 baseline MEP elicited by transcranial magnetic stimulation, followed by 50 MEP immediately after nonfatiguing exercise of the examined muscle. Postexercise MEP facilitation, expressed as percentage of baseline value, varied from 71% to 119% and from 99% to 107% in each patient, respectively, being significantly lower than our mean normal control value (268%). No differences in MEP facilitation between phases of short-circle depressive-manic disorder were revealed. Reduced postexercise facilitation was independent of the bipolar disorder phases, suggesting an invariable underlying association of the psychiatric pathophysiological mechanisms to impaired cortical excitability.