Nano-HPLC-MS analysis of phospholipids in cerebrospinal fluid of Alzheimer's disease patients--a pilot study.

There is emerging evidence that lipids play an important role in many neurodegenerative processes, for example in Alzheimer's disease (AD). Although different lipid alterations in the AD brain have been reported, there have only been very few investigations of lipid changes in the cerebrospinal fluid (CSF). Recent developments in mass spectrometry (MS) have enabled fast and sensitive detection of lipid species in different biological matrixes. In this study we developed an on-line HPLC-MS method for phospholipid profiling in the CSF based on nano-HPLC separation using an Amide column and detection with electrospray (ESI) quadrupole-time of flight (QTOF) MS. We achieved good separation, reproducibility, and sensitivity in monitoring of the major phospholipid classes, phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylinositol (PI), and sphingomyelin (SM) in CSF. To emphasize the applicability of the method, a pilot study was performed on a group of CSF samples (N = 16) from individuals with probable AD and non-demented controls. We observed a statistically significant increase of SM levels (24.3 ± 2.4%) in CSF from probable AD individuals vs. controls. Our findings indicate that SM levels in the CSF could potentially provide a new lead in AD biomarker research, and show the potential of the method for disease-associated CSF phospholipid screening.

Distinguishing Parkinson's disease and essential tremor with transcranial sonography.

OBJECTIVES: Until today there is no reliable test that can clearly distinguish Parkinson's disease (PD) from the essential tremor (ET). Our aim was to determine the usefulness of the transcranial sonography (TCS) in the differential diagnosis of the PD and ET as well as the interobserver reliability for this method. METHODS: Transcranial sonography of substantia nigra and clinical examination were performed on 80 PD patients, 30 ET patients, and 80 matched controls by two independent physicians. RESULTS: Bilateral SN hyperechogenicity over the margin of 0.20 cm(2) was found in 91% of PD patients, 10% of healthy subjects, and in 13% patients with ET. Interobserver agreement for this method was significant (Student's t-test, P = 1.000). CONCLUSIONS: Substantia nigra hyperechogenicity on TCS is a highly specific finding of PD, where in healthy individuals or in ET patients, it might correspond to an increased risk of developing PD later in life or might also be because of the impairment of nearby area of nucleus ruber in ET patients, as suggested by positron emission tomography studies. TCS may serve as a practical and sufficiently sensitive neuroimaging tool in PD diagnoses and in distinguishing it from ET; its repeatability and accuracy might add to its practical value.

Frequency and characteristics of emotional disorders in patients after ischemic stroke

Emotional disturbances in stroke patients may unfavorably affect the process of rehabilitation and long-term outcome of the disease. The aim of the study was to assess the prevalence of emotional disturbances and their characteristics in our stroke patients, according to hemispheric lateralization of cerebral lesion (as recorded by CT), patient sex and grade of neurological handicap (as assessed by Rankin scale).The study included 50 patients (29 men and 21 women, mean age 65.52 ± 7.07 and64.62 ± 11.83 years, respectively) who had suffered ischemic stroke 3 weeks to 6 months before the study. The Crown-Crisp experience index which consists of six scales: scales of anxiety, phobia, obsession, somatization, depression and hysteria, were used for detection of emotional disturbances. Results showed a high prevalence of emotional disturbances in the study group. Depression was most common (36 of study patients), followed by generalized anxiety(n=29) and phobic disturbances (n=33). According to hemispheric lateralization of the cerebral lesion, a more intense emotional response was found in case of right hemispheric lesions, however, the difference was statistically significant only on the scale of somaticized anxiety (p<0.05). According to sex, a more intense emotional response was recorded in women. The difference being statistically significant on the scales of anxiety(p<0.05), depression (p<0.05) and phobia (p<0.01). An increasing tendency in the prevalence of emotional disturbances was observed with the increasing severity of neurologic deficit (p<0.05) (AU)

The secular trend in the incidence of hemorrhagic stroke in the region of Osijek, Eastern Croatia in the period 1988-2000--a hospital based study.

The purpose of the study was to establish the possible environmental influences in the observed peculiar rising and falling oscillations in the numbers of hemorrhagic stroke (HS) in Eastern Croatia (region of Osijek) during the last thirteen-years' period (1988-2000). In this period 1,222 HS were registered and treated. A constant increase in the incidence of HS was observed, from 60 (in 1988) to 139 (in 1998), with an average annual proportion of 16.5% of all stroke cases. A sharp increase in proportion of HS in total stroke incidence was recorded during the war in Croatia (1991-1995), with a peak incidence of 27.4% in 1993. Typical hypertensive intracerebral hemorrhage (ICH) was the most common (57.1%), atypical ICH occurred in 26.4%, subarachnoid hemorrhage (SAH) in 16.5%. Analysis of the annual number of hypertensive-ICH and SAH disclosed peculiar rising and falling oscillations. These variations were in correlation with heavy living conditions. During the war-period the SAH incidence sharply rose. Immediately after the war it suddenly decreased. The authors named this phenomenon a "pool depletion", supposing the relatively stable proportion of the bearers of aneurysms in population. The observed variations seem to be the consequence of the war stress and other negative psychosocial and economic factors in post-war period, which increases the risk for SAH and typical hypertensive-ICH through complex pathophysiological mechanisms.

Presynaptic vesicles, exocytosis, membrane fusion and basic physical forces.

The theoretical hypothesis is presented trying to explain the vesicle release from presynaptic nerve ending and membrane fusion. This theoretical concept implies only essential physical forces such as electrostatic force and surface tension force. Transmembrane resting potential of approximately -70 to -80 mV means that the intracellular fluid is electronegative in comparison with extracellular one. In this concept it is supposed that the inner and outer lipid layer of the membrane also have different electrostatic charges. Presynaptic vesicles are made from cell membrane by endocytic process through which the vesicle loses the contact with cell membrane. Also, during the endocytic process, the inner lipid layer of the cell membrane becomes the outer lipid layer of presynaptic vesicle and vice versa. During the resting phase, equally charged lipid layers of presynaptic vesicle and cell membrane repel each other, but during the action potential, differently charged lipid layers strongly attract each other, bringing the presynaptic vesicle and cell membrane in close contact. Immediately thereafter, the surface tension forces open the pore and fuse both membranes trying to minimize the area of the contact between water fluids (extra and intracellular fluid) and lipid fluids (lipid membrane bilayer). Since only fundamental physical forces are involved in this process, it could be very fast, effective and almost inexhaustible. Similar mechanisms could be responsible for all exocytic processes and all membrane fusion processes in the cells.

Tuberculosis and alcoholism in Croatia.

Alcoholism and tuberculosis represent very difficult problem in treatment because both illnesses require complex treatment which cannot be adequately obtained on classic wards. Tuberculosis in the last 50 years showed the constant decline of incidence--in Croatia (4,500,000 inhabitants) from approximately 20,000 to approximately 2000 cases per year. This study compares preliminary results of AUDIT (Alcohol Use Disorders Identification Test) in the group of tuberculosis patients with the diagnosis of alcoholism (group 1) and in the group of tuberculosis patients who had not the diagnosis of alcoholism (group 2) and who all were admitted to the Hospital for pulmonary diseases and tuberculosis Klenovnik where is an organised unique ward management for tuberculosis and alcoholic patients functioning for the last 25 years on the basic principles of therapeutic community. Their average AUDIT score was 25.44--very high, and all patients scored above 8, what indicated on harmful and hazardous drinking. Additionally, in the group of patients admitted for hospital treatment of tuberculosis without diagnosis alcoholism there were more than 50% of harmful or hazardous alcohol drinkers who ranged 8 or more on AUDIT score, what could indicate that modern identification of hazardous drinking (for example using AUDIT) should be used in all pulmonary and tuberculosis wards. Patients for whom diagnostic procedure shows that they drink harmfully or hazardously should be treated simultaneously for tuberculosis and alcohol use disorders. Because of hidden alcohol problems the group sociotherapy should be organised for all patients at all wards for treatment of tuberculosis in a common and special alcohologic way.

[Cotrimoxazole in the treatment of Wegener's vasculitis (case report)]. / Kotrimoksazol u lijecenju Wegenerova vaskulitisa (prikaz bolesnika).

A female patient suffering from Wegener's vasculitis of upper respiratory tract, lungs, kidney and skin, is presented. The diagnosis has been established on the basis of clinical course, histopathologic examination of lung tissue obtained by transbronchial lung biopsy and positive serum antineutrophil cytoplasmatic antibodies (ANCA). The patient was successfully treated with trimethoprim-sulfamethoxasole (daily dose 320 + 1600 mg) leading to complete clinical remission. The course of disease and the effects of treatment were strongly drug-dose-related. We emphasized some diagnostic difficulties and new trends in the therapy of Wegener's granulomatosis and other ANCA positive vasculitides. Possible mechanisms of the trimethoprim-sulfamethoxasole efficiency in Wegener's granulomatosis are also discussed.

Electrostatic attraction: the driving force for the presynaptic vesicle-cell membrane fusion.

In this theoretical conception, it is suggested that the driving force for presynaptic vesicle-cell membrane fusion during action potential that results with the neurotransmitter release is the electrostatic attraction of the differently charged outer lipid layer of the presynaptic vesicle membrane and the inner lipid layer of the cell membrane. All other previously described processes that take place during presynaptic vesicle-cell membrane fusion could serve to modulate the basic electrostatic process presented here.

Utilization of antituberculosis drugs expressed in defined daily doses in Klenovnik Hospital in the period between 1983 and 1987.

In this paper the utilization of antituberculosis drugs was analyzed in defined daily doses per 1000 bed-days (DDD/1000 BD) in the Hospital for Pulmonary Diseases and Tuberculosis, Klenovnik, from 1983 to 1987. The utilization of these drugs increased from 894 DDD/1000 BD in 1983 to 1112 DDD/1000 BD in 1984, and then decreased to 1077 DDD/1000 BD in 1986; but in 1987 it again increased to 1270 DDD/1000 BD. During the research period the following drugs were prescribed: ethambutol, rifampin, pyrazinamide, streptomycin, isoniazid, and two combinations of drugs: a combination of isoniazid with pyridoxine and a combination of ethambutol, isoniazid and pyridoxine. For the whole of that period the use of ethambutol, isoniazid with pyridoxine and rifampin made up more than 85% of the general utilization of antituberculosis drugs, while other drugs were prescribed in lesser quantities. The data presented indicate that tuberculosis in Klenovnik hospital was in most cases treated with ethambutol, the combination of isoniazid with pyridoxine, and rifampin.