Integrative Metabolomic Analysis of Serum and Selected Serum Exosomal microRNA in Metastatic Castration-Resistant Prostate Cancer.

Metastatic castration-resistant prostate cancer (mCRPC) remains a lethal disease due to the absence of effective therapies. A more comprehensive understanding of molecular events, encompassing the dysregulation of microRNAs (miRs) and metabolic reprogramming, holds the potential to unveil precise mechanisms underlying mCRPC. This study aims to assess the expression of selected serum exosomal miRs (miR-15a, miR-16, miR-19a-3p, miR-21, and miR-141a-3p) alongside serum metabolomic profiling and their correlation in patients with mCRPC and benign prostate hyperplasia (BPH). Blood serum samples from mCRPC patients (n = 51) and BPH patients (n = 48) underwent metabolome analysis through 1H-NMR spectroscopy. The expression levels of serum exosomal miRs in mCRPC and BPH patients were evaluated using a quantitative real-time polymerase chain reaction (qRT-PCR). The 1H-NMR metabolomics analysis revealed significant alterations in lactate, acetate, citrate, 3-hydroxybutyrate, and branched-chain amino acids (BCAAs, including valine, leucine, and isoleucine) in mCRPC patients compared to BPH patients. MiR-15a, miR-16, miR-19a-3p, and miR-21 exhibited a downregulation of more than twofold in the mCRPC group. Significant correlations were predominantly observed between lactate, citrate, acetate, and miR-15a, miR-16, miR-19a-3p, and miR-21. The importance of integrating metabolome analysis of serum with selected serum exosomal miRs in mCRPC patients has been confirmed, suggesting their potential utility for distinguishing of mCRPC from BPH.

Surgical treatment of severe dumping syndrome.

Dumping syndrome is a common post-operative complication following gastric surgery. Clinically, severe dumping can be a serious medical condition with a negative impact on the patient's life. In our case report, we present a case of refractory dumping syndrome which developed after laparoscopic subtotal gastrectomy with gastrojejunoanastomosis due to massive gastroptosis with stomach evacuation problems. Conservative gastroenterology treatment was not successful. Due to the progression of weight loss and life-threatening hypoglycaemia, the decision for surgical treatment was made. After the corrective gastro-duodenal and jejuno-jejunal anastomoses, all clinical symptoms resolved completely. With regard to the presented case, we discuss the common treatment options for dumping syndrome: the standard recommendations for dietary habits, pharmacological treatment and finally the surgery and its pitfalls. Due to the absence of randomized trials and guidelines, every patient should be treated in a personalized way.

Altered hypothalamic metabolism in early multiple sclerosis - MR spectroscopy study.

Multiple sclerosis (MS) is a disease characterized by overlapping processes of neuroinflammation and neuro-axonal degeneration. Disturbances of the hypothalamo-pituitary axis in MS are supposed to modulate neuroinflammatory circuits, however, there is insufficient knowledge about the hypothalamic metabolism alterations in early MS. This 1H MRS study performed on a 1.5 T MR-scanner was focused on the hypothalamus of 31 pre-treatment patients after their first clinical MS episode/s, compared to 31 healthy controls. The metabolite ratios of N-acetyl-aspartate &N-acetyl-aspartyl-glutamate (tNAA), glutamate & glutamine (Glx), myo-Inositol (mIns), choline- and creatine-containing compounds (tCho, tCr) were further correlated with the Expanded Disability Status Scale (EDSS). In the hypothalamus of early MS patients compared to controls, we found decreased tNAA/tCr and increased tCho/tNAA, mIns/tNAA, Glx/tCr, and Glx/tNAA. In addition, tCho/tNAA, Glx/tNAA, and mIns/tNAA were positively and tNAA/tCr was negatively correlated with EDSS. Results suggest that the decline of the tNAA ratio, indicating neuro-axonal dysfunction in the hypothalamus, may be linked with glutamate excitotoxicity. Excessive glutamate concentrations may cause microglial activation and myelinated tracts degradation with subsequent gliosis, paralleled by increased mIns and tCho ratios. This indicates that glutamate excitotoxicity can play an important role in MS from its earliest stages.

Why the Gold Standard Approach by Mammography Demands Extension by Multiomics? Application of Liquid Biopsy miRNA Profiles to Breast Cancer Disease Management.

In the global context, the epidemic of breast cancer (BC) is evident for the early 21st century. Evidence shows that national mammography screening programs have sufficiently reduced BC related mortality. Therefore, the great utility of the mammography-based screening is not an issue. However, both false positive and false negative BC diagnosis, excessive biopsies, and irradiation linked to mammography application, as well as sub-optimal mammography-based screening, such as in the case of high-dense breast tissue in young females, altogether increase awareness among the experts regarding the limitations of mammography-based screening. Severe concerns regarding the mammography as the "golden standard" approach demanding complementary tools to cover the evident deficits led the authors to present innovative strategies, which would sufficiently improve the quality of the BC management and services to the patient. Contextually, this article provides insights into mammography deficits and current clinical data demonstrating the great potential of non-invasive diagnostic tools utilizing circulating miRNA profiles as an adjunct to conventional mammography for the population screening and personalization of BC management.

Increased glutamate and deep brain atrophy can predict the severity of multiple sclerosis.

OBJECTIVE: In multiple sclerosis (MS), deep grey matter (DGM) atrophy has been recognised as a crucial component of the disease that presents early and it has been associated with disability. Although the precise mechanism underlying grey matter atrophy is unknown, several hypotheses have been postulated. Our previous research pointed to correlations of hypothalamic metabolic alterations with clinical outcomes of MS, therefore we decided to further test the relationship of these alterations with DGM atrophy. METHODS: We used 1H-Magnetic Resonance spectroscopy (1H-MRS) of the hypothalamus to test its metabolites in 26 patients with RRMS and 22 healthy age-matched controls. DGM atrophy was evaluated by simple planimetry of third ventricular width on the hypothalamic level (3VW) in T1 weighted MRI pictures. Metabolite ratios of N-acetyl aspartate (NAA), choline (Cho), glutamate and glutamine (Glx), myo-inositol (mIns) and creatine (Cr) were correlated with Multiple Sclerosis Severity Scale (MSSS) and 3VW. RESULTS: Metabolite concentrations were compared between patients and controls using multiple regression models allowing for age, 3VW and metabolites. It revealed that the only relevant predictors of MSSS were 3VW and Glx/NAA. At a significance level of P<0.05, a unit increase of 3VW was associated with a 0.35 increase of MSSS, for a typical value of Glx/NAA; P value 0.0039. A unit increase of Glx/NAA was associated with a 0.93 increase of MSSS, for a typical value of atrophy; P value 0.090. There were significant linear correlations between Glx/Cr and MSSS, Glx/NAA and MSSS, and between mIns/NAA and 3VW. CONCLUSIONS: The results suggest that both NAA and Glx are associated with neurodegeneration of hypothalamic DGM and severe disease course. Glx related 1H-MRS parameters seem to be superior to other metabolites in determining disease burden, independently of otherwise powerful 3VW planimetry. Significantly increased mIns/NAA in MS patients compared to controls point to gliosis, which parallels the atrophy of hypothalamic DGM.

Contribution to the Diagnosis and Treatment of Life-Threatening Parasitosis Caused by the Parasite Echinococcus multilocularis.

Echinococcosis is a serious parasitic disease that ends lethally in 95% of untreated infected patients. It was first diagnosed in Slovakia in the year 2000. It is caused by the larval stage of a tapeworm belonging to the genus Echinococcus, which was assigned to the group "A" of zoonoses in the year 2004. The number of new infections is rising because of increasing percentage of infected red foxes (Vulpes vulpes). Early and accurate diagnosis of infections with this parasite is essential for proper initiation of adequate therapy. Thanks to professional multidisciplinary efforts and new laboratory procedures, the number of correctly diagnosed cases has increased. Antimicrobial therapeutic approaches can lead to improved quality of life and better prognosis even if radical surgery is refused by the patient.

Congenital complete and partial absence of the left pericardium.

Congenital absence of pericardium is a rare malformation. We report 2 young patients with a diagnosis of congenital absence of the pericardium. The posteroanterior view of the chest X-ray showed displacement of the left cardiac border into the left hemithorax. Unusual acoustical windows and abnormal cardiac and septal motion during echocardiography suggested the diagnosis of congenital absence of pericardium. Magnetic resonance imaging definitive confirmed diagnosis of congenital absence of pericardium.

Treatment of cervical internal carotid artery spontaneous dissection with pseudoaneurysm and unilateral lower cranial nerves palsy by two silk flow diverters.

Internal carotid artery (ICA) lesions in the parapharyngeal space (a dissection and a pseudoaneurysm) may present as isolated lower cranial nerves (IX, X, XI, and XII) palsy (Collet-Sicard syndrome). Some arteriopathies such as fibromuscular dysplasia and tortuosity make a vessel predisposed to dissection. Extreme vessel tortuosity makes the treatment by a stent graft impossible. Two Silk stents were used in a 46 year-old man with left lower cranial nerves (IX-XII) palsy for the treatment of left ICA spontaneous dissection with pseudoaneurysm. A follow-up angiogram 5 months later confirmed pseudoaneurysm thrombosis and patency of the left ICA. The patient recovered completely from the deficits.

Flow changes after endovascular treatment of a wide-neck anterior communicating artery aneurysm by using X-configured kissing stents (cross-kissing stents) technique.

Endovascular treatment for a wide-neck anterior communicating artery (AcomA) aneurysm remains technically challenging. Stent-assisted embolization has been proposed as an alternative of treatment of complex aneurysms. The X-configuration double-stent-assisted technique was used to achieve successful coiling of wide-neck AcomA aneurysm. Implanted stent can alter intra-arterial flow. Follow-up angiograms 4 months later showed flow changes due to used X-technique of stents implantation and filling of the anterior cerebral artery from the opposite internal carotid artery.

Proton magnetic resonance spectroscopy in patients with early stages of amyotrophic lateral sclerosis.

INTRODUCTION: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder primarily affecting upper and lower motor neurons. Due to relative fast progression of the disease, early diagnosis is essential. Proton magnetic resonance spectroscopy ((1)H-MRS) is used for objectivization of upper motor neuron (UMN) lesions. The aim of this study was to assess the use of (1)H-MRS in the early stages of ALS. METHODS: Eleven patients with clinically definite (n=2), probable (n=7), and probable laboratory-supported (n=2) diagnosis of ALS with disease duration of less than 14 months were studied. Control group consists of 11 sex- and age-matched healthy subjects. All subjects underwent assessment of functional disability using revised ALS Functional Rating Scale (ALSFRS-R) and single-voxel (1)H-MRS examination of both precentral gyri, pons, medulla oblongata, and occipital lobe. Spectra were evaluated with LCModel software. RESULTS: The mean disease duration was 6.5 ± 3.5 months. The median ALSFRS-R was 42. Significant decrease between patient and control groups was found in the NAA/Cre ratio in the left and right precentral gyri (p=0.008, p=0.040). Other metabolite ratios in other areas did not show significant differences. Total ALSFRS-R score weakly positively correlated with NAA/Cre ratio in the left precentral gyrus (p=0.047). CONCLUSIONS: (1)H-MRS is sensitive to detect metabolic changes caused by neurodegeneration processes during ALS and can be used for detection of UMN dysfunction. These MRS changes in the early stages of ALS are most prominent in motor cortex.

Bursts of 15-30 Hz oscillations following noxious laser stimulus originate in posterior cingulate cortex.

Previous EEG studies reported the presence of synchronised 15-30 Hz oscillations in vertex electrodes following innocuous somatosensory stimulation and noxious laser stimulation. The purpose of the present study was to analyse the sources of poststimulus increases of 15-30 Hz oscillations during noxious laser stimulation of four different body regions and to compare the sources of the poststimulus synchronisation with the sources of the N2 component of laser-evoked potential (LEP). In 10 healthy subjects, moderately painful laser stimuli were applied to the dorsum of the right hand, dorsum of the right foot, right groin, and right side of the face. EEG data, recorded from 111 scalp sites, were analysed using event-related desynchronisation method and source dipole analysis. A profound amplitude increase of 15-30 Hz oscillations peaking 1-2 s after noxious laser stimulation was found during stimulation of each body part. The sources of these oscillations were located in the dorsal posterior cingulate cortex and showed no somatotopic arrangement. The sources of the N2-LEP component were located in the anterior mid-cingulate cortex 25-30 mm rostral to the sources of 15-30 Hz oscillations. The amplitude of the poststimulus synchronisation of 15-30 Hz oscillations correlated (P<0.05) with the amplitude of N2-LEP component. Results show that noxious laser stimuli induce bursts of 15-30 Hz oscillations in the posterior cingulate cortex. The poststimulus increases of 15-30 Hz oscillations may stand for transient cortical inhibition possibly aiding temporary suppression of motor programs that have been primed by noxious stimulation.

Broken esophageal stent successfully treated by interventional radiology technique.

Esophageal stent fractures occur quite rarely. A 61-year-old male patient was previously treated for rupture of benign stenosis, occurring after dilatation, by implanting an esophageal stent. However, a year after implantation, the patient suffered from dysphagia caused by the broken esophageal stent. He was treated with the interventional radiology technique, whereby a second implantation of the esophageal stent was carried out quite successfully.

[Cyst of Echinococcus etiology localised in the spleen]. / Cysta echinokokovej etiológie lokalizovaná v slezine.

Cystic form in the spleen was diagnosed accidentally in a 44 years old patient during ultrasonography of the abdomen. It reached 6.0 cms in diameter. The mass was localised superficially, closely beneath the capsule of the spleen. The diagnose was confirmed serologically. Because of the localisation and the size of the cyst, the surgical approach - splenectomy - was indicated and performed.

A novel JAK2 exon 12 mutation identified in the retrospective analysis of paraffin-embedded tissues of polycythemia vera patients.

The aim of the study was to screen formalin-fixed, paraffin-embedded tissues of polycythemia vera patients for the presence of JAK2(V617F) and JAK2 exon 12 mutations. Of 64 cases, 60 were positive for JAK2(V617F) mutation using allele-specific polymerase chain reaction (PCR). Using modified allele-specific PCR, samples of 4 JAK2(V617F)-negative patients were analyzed for the presence of JAK2 exon 12 mutations. In one case, we found a PCR product matching with allele-specific primer, which was designed to detect the N542-E543del mutation. Surprisingly, in the result sequence we have detected another recently described mutation, R541-E543delinsK. In the other 3 JAK2(V617F)-negative patients, allele-specific PCR for the detection of JAK2 exon 12 mutations did not yield any product. However, in 1 case, the sequencing of JAK2 exon 12 PCR product revealed a novel mutation, H538-K539delinsF. We confirmed that paraffin-embedded tissues represent a valuable source of DNA, which can be used in diagnostics of both JAK2 exon 12 and exon 14 mutations and we described 1 novel JAK2 exon 12 mutation.

Short reflex expirations (expiration reflexes) induced by mechanical stimulation of the trachea in anesthetized cats.

Fifty spontaneously breathing pentobarbital-anesthetized cats were used to determine the incidence rate and parameters of short reflex expirations induced by mechanical stimulation of the tracheal mucosa (ERt). The mechanical stimuli evoked coughs; in addition, 67.6% of the stimulation trials began with ERt. The expiration reflex mechanically induced from the glottis (ERg) was also analyzed (99.5% incidence, p < 0.001 compared to the incidence of ERt). We found that the amplitudes of abdominal, laryngeal abductor posterior cricoarytenoid, and laryngeal adductor thyroarytenoid electromyograms (EMG) were significantly enhanced in ERg relative to ERt. Peak intrathoracic pressure (esophageal or intra-pleural pressure) was higher during ERg than ERt. The interval between the peak in EMG activity of the posterior cricoarytenoid muscle and that of the EMG of abdominal muscles was lower in ERt compared to ERg. The duration of thyroarytenoid EMG activity associated with ERt was shorter than that in ERg. All other temporal features of the pattern of abdominal, posterior cricoarytenoid, and thyroarytenoid muscles EMGs were equivalent in ERt and ERg.In an additional 8 cats, the effect of codeine administered via the vertebral artery was tested. Codeine, in a dose (0.03 mg/kg) that markedly suppressed cough did not significantly alter either the incidence rate or magnitudes of ERt.In the anesthetized cat the ERt induced by mechanical stimulation of the trachea was similar to the ERg from the glottis. These two reflex responses differ substantially only in the frequency of occurrence in response to mechanical stimulus and in the intensity of motor output.

Functional magnetic resonance imaging of cerebral activation during spinal cord stimulation in failed back surgery syndrome patients.

Spinal cord stimulation (SCS) consisting of electrical stimulation of the dorsal spinal cord using epidural electrodes has been shown to relieve chronic neuropathic pain. To analyze the cerebral activation patterns related to SCS, and to evaluate the effects of SCS on the processing of acute experimental pain, we performed functional magnetic resonance imaging (fMRI) on eight patients suffering from failed back surgery syndrome who were also being treated with SCS for severe pain in their legs and lower back. Three types of stimulation were used, each lasting 36s: (i) SCS, (ii) heat pain (HP) applied to the leg affected by neuropathic pain, and (iii) simultaneous HP and SCS. During SCS, we found increased activation of the medial primary sensorimotor cortex somatotopically corresponding to the foot and/or perineal region, contralateral posterior insula, and the ipsilateral secondary somatosensory cortex (S2). Decreased activation was seen in the bilateral primary motor cortices and the ipsilateral primary somatosensory cortex corresponding to the shoulder, elbow and hand. Compared to separately presented HP and SCS, simultaneous HP and SCS showed statistically significant activation of the bilateral inferior temporal cortex and the ipsilateral cerebellar cortex. The activation of the primary motor cortex, insula and S2 during SCS may directly interfere with the processing of neuropathic pain. When SCS is associated with heat pain, the paralimbic association cortex and cerebellum show activation exceeding the sum of activations resulting from separate SCS and heat pain stimulation. The explanation of this could possibly rest with the continuous comparisons of simultaneous pain and somatosensory sensations occurring in a single dermatome.

Skeletal fluorosis from the point of view of an occupational exposure to fluorides in former Czechoslovakia.

Electrolytic production of aluminium in former Czechoslovakia started in the year 1953 in the Ziar valley in the central Slovakia. However, till 1995 the hygienic conditions for health protection were not met in the factory. It underwent a reconstruction afterwards.The authors demonstrate cases of occupational skeletal fluorosis (currently rare in Europe) in 14 metallurgists which were all disclosed in foundry workers in Ziar nad Hronom as to the year 2005. The occupational disease was diagnosed after 17.7 ± 7.67 years (x±SD) of exposure in the foundry.The authors describe the clinical conditions, haematological and biochemical tests (decreased level of ionising calcium was found in serum). The content of fluorides in urine was increased (254.4±130.95 µmol/l). The average age of patients at the time of recognition of the professional etiology of the disease was 57.93±7.95 years. Eight patients were older than 60 years. Skeletal abnormalities were evaluated by using X-ray skiagraphy, estimating the Stage I-III of the skeletal fluorosis. Typically an increase of bone density was found, the compact part of long bones was coarsed, there were calcifications of the interosseous membrane between radius and ulna and some ossifications of the sacrospinal and sacrotuberous ligaments. Twelve patients suffered sensorimotor polyneuropathy of extremities, chronic bronchitis was found in 6 patients (two of them were smokers).The last occupational case was registered in the year 2001. The authors assume that aluminium production with modern technology of better safety and protection of health of workers is the key which will make the skeletal fluorosis the history in the Czech and Slovak Republic.

Effects of spinal cord stimulation on the cortical somatosensory evoked potentials in failed back surgery syndrome patients.

OBJECTIVE: To evaluate the functional activation of the somatosensory cortical regions in neuropathic pain patients during therapeutic spinal cord stimulation (SCS). METHODS: In nine failed back surgery syndrome patients, the left tibial and the left sural nerves were stimulated in two sessions with intensities at motor and pain thresholds, respectively. The cortical somatosensory evoked potentials were analyzed using source dipole analysis based on 111 EEG signals. RESULTS: The short-latency components of the source located in the right primary somatosensory cortex (SI: 43, 54 and 65ms) after tibial nerve stimulation, the mid-latency SI component (87ms) after sural nerve stimulation, and the mid-latency components in the right (approximately 161ms) and left (approximately 168ms) secondary somatosensory cortices (SII) were smaller in the presence of SCS than in absence of SCS. The long-latency source component arising from the mid-cingulate cortex (approximately 313ms) was smaller for tibial and larger for sural nerve stimuli during SCS periods compared to periods without SCS. CONCLUSIONS: SCS attenuates the somatosensory processing in the SI and SII. In the mid-cingulate cortex, the effect of SCS depends on the type of stimulation and nerve fibers involved. SIGNIFICANCE: Results suggest that the effects of SCS on cortical somatosensory processing may contribute to a reduction of allodynia during SCS.

Source imaging of the cortical 10 Hz oscillations during cooling and warming in humans.

Primary cold and warm afferent fibers show a robust overshoot in their firing during periods of temperature change, which subsides during tonic thermal stimulation. Our objective was to analyze cortical activation, on a scale of hundreds of milliseconds, occurring during the process of dynamic cooling and warming, based on an evaluation of the amplitude changes seen in 10 Hz electroencephalographic oscillations. Eleven right-handed subjects were exposed to innocuous cold ramp stimuli (from 32 degrees C to 22 degrees C, 10 degrees C/s) and warm ramp stimuli (32 degrees C to 42 degrees C, 10 degrees C/s) on the thenar region of their right palm, using a contact thermode. EEG was recorded from 111 scalp sites, and the 10 Hz current source densities were modeled using low-resolution electromagnetic tomography. During cooling, the earliest amplitude decreases of 10 Hz oscillations were seen in the contralateral posterior insula and secondary somatosensory cortex (SII), and the premotor cortex (PMC). During warming, the earliest events were only observed in the PMC and occurred approximately 0.7 s later than during cooling. Linear regression analysis between 10 Hz current source densities and temperature variations revealed cooling-sensitive activation in the bilateral posterior insula, PMC and the anterior cingulate cortex. During warming, the amplitude of 10 Hz oscillations in the PMC and posterior insula correlated with stimulus temperature. Dynamic thermal stimulation activates, in addition to the posterior insula and parietal operculum, the lateral PMC. The activation of the anterior cingulate cortex during cooling may aid in the anticipation of the cold temperature end-point and provide continuous evaluation of the thermal stimulus.

Somatosensory-evoked potentials are influenced differently by isometric muscle contraction of stimulated and non-stimulated hand in humans.

Chronic pain is associated with motor dysfunctions, and stimulation of the motor cortex has been shown to alleviate chronic pain. Recently Nakata et al. [Pain 107 (2004) 91-98] showed differentiated patterns of neuromagnetic evoked fields following painful laser stimulation during phasic movements of stimulated and non-stimulated hand. Phasic movements and static contractions differ in their functional activation of the motor cortices. Therefore, we decided to analyze the effects of isometric contractions of intrinsic right and left hand muscles on cortical sources of somatic-evoked potentials related to a painful galvanic stimulation of the right middle finger. Using spatio-temporal source dipole analysis of 111 electroencephalographic signals in 10 right-handed men, source activities were evaluated in the left primary somatosensory cortex (S1), left (S2(L)) and right (S2(R)) secondary somatosensory cortex, anterior cingulate cortex (ACC) and posterior cingulate cortex (PCC). Ipsilateral hand muscle contraction was associated with a decrease of source activity in S1 and with subsequent increases in S2(L) and the PCC. Contralateral hand muscle contraction was accompanied by a decrease of source activity in bilateral S2 cortices followed by decreases in the S1 and anterior cingulate cortex. Results suggest early suppression of source activity in S1 during ipsilateral hand muscle contractions and in bilateral S2 during contralateral hand muscle contractions.