Taurine in plasma and CSF: a study in healthy male volunteers

In order to explore the interrelationship between plasma and cerebrospinal fluid taurine concentrations, three consecutive 6-ml fractions of cerebrospinal fluid were drawn from 30 healthy male volunteers in the early morning after 8 h in the fasting condition. Repeated plasma samples were drawn over 24 h the day before lumbar puncture. Taurine in plasma and cerebrospinal fluid was determined by high performance liquid chromatography. The subjects were categorized as extensive or poor metabolizers with respect to the cytochrome P450 2D6 genotype. The taurine cerebrospinal fluid/plasma ratio at 8 a.m. was negatively influenced by the plasma taurine concentration at 4 p.m. the previous day. It was also negatively influenced by body mass index and positively by the intraspinal pressure. Three poor metabolizers of cytochrome P450 2D6 had higher plasma taurine areas under the curve than 27 extensive metabolizers. Hypothetically, cytochrome P450 2D6 influences the transport of taurine across the blood–brain barrier.     

Usefulness of McRAPD for typing and importance of biofilm production in a case of nosocomial ventriculoperitoneal shunt infection caused by <Emphasis Type="Italic">Candida lusitaniae</Emphasis>

A case report of ventriculoperitoneal shunt infection caused by Candida lusitaniae in a 6-year-old patient with cerebral astrocytoma and obstructive hydrocephalus is presented briefly with emphasis on the course of antifungal treatment. Seven isolates recovered subsequently from the cerebrospinal fluid were studied retrospectively. To confirm identity, isolates were typed using pulsed-field gel electrophoresis and melting curve of random amplified polymorphic DNA (McRAPD). Further, the ability to form biofilm and its susceptibility to systemic antifungals were evaluated. Using McRAPD, identity of C. lusitaniae isolates showing slight microevolutionary changes in karyotypes was undoubtedly confirmed; successful application of numerical interpretation of McRAPD for typing is demonstrated here for the first time. The strain was also recognized as a strong biofilm producer. Moreover, minimum biofilm inhibitory concentrations were very high, in contrast to low antifungal minimum inhibitory concentrations of isolates. It can be concluded that McRAPD seems to be a simple and reliable method not only for identification but also for typing of yeasts. A ventriculoperitoneal shunt colonized by C. lusitaniae was revealed as the source of this nosocomial infection, and the ability of the strain to form biofilm on its surface likely caused treatment failure.     

神经外科颅脑手术患者发生术后颅内感染的危险因素及应用中医外科"托法"效果分析

摘要 目的：探讨神经外科颅脑手术患者术后发生颅内感染的危险因素，并研究中医外科"托法"辅助治疗颅内感染的临床疗效。方法：回顾性分析2017年1月到2022年12月在我院神经外科进行颅脑手术的260例患者临床资料，分析影响患者发生术后颅内感染的影响因素。发生术后发生颅内感染的患者在常规抗感染治疗的基础上加用中医"托法"治疗，分析其临床治疗疗效、抗感染治疗后脑脊液白细胞计数、蛋白含量和中性粒细胞，以及血清白介素-8（IL-8），超敏c反应蛋白（hs-CRP）和降钙素（PCT）。结果：260例神经外科颅脑手术患者，术后出现颅内感染的患者有21例，术后颅内感染发生率为8.08%。多因素Logistic回归分析显示：手术时间、脑脊液分流术、脑室外引流以及脑脊液漏是影响神经外科颅脑手术患者术后是否发生颅内感染的独立影响因素。经中医外科"托法"辅助抗感染治疗后，21例术后颅内感染患者脑脊液白细胞计数、蛋白含量和中性粒细胞均较治疗前显著降低（P<0.05），并且血清IL-8、hs-CRP和 PCT均较治疗前降低（P<0.05）。21例术后颅内感染患者治疗总有效率、抗感染治疗时间和总费用分别为90.48%、（11.43±1.57）天和（7571.68±2541.29）元。结论：手术时间、脑脊液分流术、脑室外引流以及脑脊液漏是影响神经外科颅脑手术患者术后是否发生颅内感染的独立影响因素，中医外科"托法"可用于术后颅内感染患者抗感染治疗。     

Relation entre réaction géotropique et évolution du statenchyme dans la racine d'Asperge

Relationship between the Geotropic Response and the Evolution of the Statenchyma in Roots of Asparagus officinalis. The evolution of the statenchyma in roots of Asparagus of ficinalis seedlings, grown in obscurity, was followed during the first 17 days. After 7 days of etiolation, a decrease of both the average diameter of the amyloplasts and the average number of these organelles was observed in the central root cap cells. If the seedlings were illuminated (with white light) from the 7th day, the average number of statoliths increased rapidly in the statocytes. The volume of these organelles undergoes the same variation in etiolated and in illuminated plants. The initial rate of curvature (V_i) of the roots (stimulated in a horizontal position) and the volume of amyloplasts (V_ac) in their caps were analysed as a function of the time of germination in obscurity (from the 8th to the 17th day). It was found that V_i increased as a linear function of the logarithm of V_ac, which confirms that the weight of the amyloplasts of the statocytes may play a role in the geotropic stimulation of the roots.     

Characterization of ψM1, a virulent phage of Methanobacterium thermoautotrophicum Marburg

We have studied the biogenesis and enzymic composition of microbodies in different yeasts during adaptation of cells to a new growth environment. After a shift of cells of Candida boidinii and Hansenula polymorpha from glucose to methanol/methylamine-containing media, newly synthesized alcohol oxidase and amine oxidase are imported in one and the same organelle together with catalase; as a consequence the cells contain one class of morphologically and enzymatically identical microbodies. Similar results were obtained when Candida utilis cells were transferred from glucose to ethanol/ethylamine-containing media upon which all cells formed microbodies containing amine oxidase and catalase.However, when methanol-limited cells of H. polymorpha were transferred from media containing ammonium sulphate to those with methylamine as the nitrogen source, newly synthesized amine oxidase was incorporated only in part of the microbodies present in these cells. This uptake was confined to the few smaller organelles generally present at the perimeter of the cells, which were considered not fully developed (immature) as judged by their size. Essentially similar results were obtained when stationary phase cells of C. boidinii or C. utilis — grown on methanol and ethanol plus ammonium sulphate, respectively — were shifted to media containing (m)ethylamine as the nitrogen source. These results indicate that mature microbodies may exist in yeasts which no longer are involved in the uptake of matrix proteins. Therefore, these yeasts may display heterogeneities in their microbody population.     

Fine Structure and Histochemistry of the Choroid Plexus of the Teleost Leuciscus rutilus

Analyses of the fine structure of the posterior choroid plexus in the teleost Leuciscus rutilus and the determination of the presence and function of the enzymes acid and alkaline phosphatases, ATPase and glucose-6-phosphatase confirm similarities between these epithelial cells and the saccus dorsalis and also with the epithelial cells of the choroid plexus found in mammals. The teleost plexus cells contain coated vesicles which are derived from the plasmalemma as well as from the Golgi complex. Moreover, they contain multivesicular bodies and Iysosomes. These organelles function in the absorption of substances from the cerebrospinal fluid and in the breakdown of these substances within the cells. The investigated enzymes play an important role in the secretion of electrolytes into the cerebrospinal fluid by active membrane transport.     

Sensitive reversed-phase high-performance liquid chromatographic method for the determination of atevirdine and its N-desethyl metabolite in human saliva or cerebrospinal fluid using solid-phase extraction

A sensitive reversed-phase high-performance liquid chromatographic method for the determination of atevirdine and its primary metabolite in human saliva or cerebrospinal fluid using solid-phase extraction is described. Samples mixed with internal standard and sodium phosphate buffer were applied to an activated C₁₈ solid-phase extraction column. The reconstituted eluate was injected onto a Zorbax RX C₈ column utilizing a mobile phase of 100 mM ammonium acetate (pH 4.0)–isopropyl alcohol–acetonitrile (55:20:25, v/v/v). Fluorescence detection was employed with excitation at 295 nm and emission at 456 nm. Quantitation was achieved using peak-height ratios. The detection response curve was linear from 2 to 850 nM for atevirdine in both human saliva and cerebrospinal fluid and from 2 to 250 nM for the metabolite in human saliva. The method was utilized to analyze cerebrospinal fluid and saliva samples from clinical studies.     

Occurrence of yeast in the intestinal tract of 20 patients during treatment with bacitracin and neomycin

Summary A mycologic investigation of stool samples of 20 patients treated with bacitracin and neomycin shows increasing growth of yeasts during the therapy. Thus 90% of the patients had yeasts in the stools atter treatment, while 37% before treatment. Most of the isolated yeast species areCandida species. The technical assistance of MissAnna Helgerud is gratefully acknowledged.     

Ultrastructure changes of seedlings of <Emphasis Type="Italic">Kosteletzkya virginica</Emphasis> under waterlogging conditions

The changes in ultrastructure of leaf cell in waterlogged Kosteletzkya virginica seedlings were observed by transmission electron microscope. During waterlogging, the chloroplasts turned into a round shape and their volume was gradually reduced. Furthermore, the lamellae of thylakoids swelled, and the inclusions in chloroplast decreased. The shape of starch grains slightly changed, but their number and volume decreased, and they even disappeared in the end. On the other hand, plastoglobules enlarged and their amount enhanced gradually. The membrane and inner cristea of mitochondria gradually became unclear, while the mitochondria firstly enlarged but disappeared at the end. Similar to chloroplast, cell nucleus dwindled gradually, concentrated and tended to a round shape. Additionally, the annulate lamellae and multivesicular body occurred on the 20th day of experiment, and the cell wall distorted and twisted at the late stage of waterlogging. Consequently, these changes of organelles are the typical characteristics of the complete cell disintegration or death for Kosteletzkya virginica under long-term waterlogging.     

Electron Microscopic Study of Demyelination in an Experimentally Induced Lesion in Adult Cat Spinal Cord

Plaques of subpial demyelination were induced in adult cat spinal cords by repeated withdrawal and reinjection of cerebrospinal fluid. Peripheral cord was fixed by replacing cerebrospinal fluid available at cisternal puncture with 3 per cent buffered OsO₄. Following extirpation, surface tissue was further fixed in 2 per cent buffered OsO₄, dehydrated in ethanol, and embedded in araldite. Normal subpial cord consists mainly of myelinated axons and two types of macroglia, fibrous astrocytes and oligodendrocytes. Twenty-nine hours after lesion induction most myelin sheaths are deteriorating and typical macroglia are no longer visible. Phagocytosis of myelin debris has begun. In 3-day lesions, axons are intact and their mitochondria and neurofibrils appear normal despite continued myelin breakdown. All axons are completely demyelinated by 6 days. They lack investments only briefly, however, for at 10 and 14 days, macroglial processes appear and embrace them. These macroglia do not resemble either one of the normally occurring glia; their dense cytoplasm contains fibrils in addition to the usual organelles. It is proposed that these macroglia, which later accomplish remyelination, are the hypertrophic or swollen astrocytes of classical neuropathology. The suggestion that these astrocytes possess the potential to remyelinate axons in addition to their known ability to form cicatrix raises the possibility of pharmacological control of their expression.     

Purification of axonin-1, a protein that is secreted from axons during neurogenesis.

Using selective metabolic labelling in a compartmental cell culture system two proteins, denoted axonin-1 and axonin-2, were found to be secreted by axons of dorsal root ganglia neurons from chicken embryos. Based on its characteristic coordinates and spot morphology in two-dimensional gel electrophoresis, axonin-1 was detected in the cerebrospinal fluid and the vitreous fluid, axonin-1 was purified 476-fold to homogeneity by a four-step chromatographic procedure. The identity of the purified protein as axonin-1 was confirmed by immunological methods. Axonin-1 is a glycoprotein that subdivides into at least 16 immunologically similar isoelectric variants; their molecular weight range extends from 132 to 140 kd and their pI range from 5.3 to 6.2. In the vitreous fluid of the embryo, axonin-1 could first be detected on the embryonic day 5 and highest concentrations were measured during the second half of embryonic life; in the vitreous fluid of the adult chicken, concentrations were approximately 20 times lower. The early onset of secretion and the time course of expression suggest a role for axonin-1 in the development of the nervous system.     

Alpha-fetoprotein expression in intra- and extraembryonic fluids of developing chick embryo

The ontogeny of alpha-fetoprotein (AFP) has been studied in the chicken (from 7 days of incubation until 2 days after hatching) using (1) the two-dimensional immunoelectrophoresis technique, (2) the polyacrylamide gel electrophoresis, and (3) the high resolution two-dimensional polyacrylamide gel electrophoresis. The molecular weight of AFP was estimated at 71,000. AFP was seen as a heterogeneous population composed of four isoforms which slightly differ by their isoelectric points. Up to the 18th day of development, qualitative changes in AFP heterogeneity do not occur. Only traces of the two alkaline isoforms were observed in plasma of 2 days post-hatching chickens. AFP has been identified in allantoic and cerebrospinal fluids but is not present in amniotic fluid. At 7 days of embryonic age, all the plasma AFP species are present in cerebrospinal fluid.     

Some basic chemical components of the cerebrospinal fluid in developing chick embryos.

The development of the chloride ion, glucose and total protein concentration was investigated in the cerebrospinal fluid of 11- to 21-day-old chick embryos and compared with their development in the blood plasma. Developmental changes in the chloride concentration in the plasma and CSF were very small, but it was always higher in the CSF than in the plasma. The plasma/CSF ratio fell during development, from 0.906 in 11-day-old embryos to 0.778 at the end of incubation. The CSF glucose concentration fell up to the 19th day of incubation, but a significant increase was recorded shortly before hatching. The plasma glucose concentration rose throughout the whole of the investigated period of embryogenesis. Up to the 19th day the P/CSF ratio rose from 1.59 to 4.05 and in 21-day-old embryos fell to 2.47. The developmental increase in the plasma total protein concentration was accompanied by the reverse process in the CSF. During the second half of incubation the P/CSF ratio rose from 1.88 to 7.9 Calculation of total osmolarity from the Na+, K+, Ca2+, Cl- and glucose concentration showed permanent hyperosmolarity of the CSF compared with the plasma. The P/CSF ratio was maintained within limits of 0.94 to 0.98.     

Unchanged levels of interleukins, neopterin, interferon-γ and tumor necrosis factor-α in cerebrospinal fluid of patients with dementia of the Alzheimer type

Several histopathological studies suggest that amyloidogenesis in dementia of the Alzheimer type is accompanied by activated glia and glia-derived cytokines, leading to chronic, self-propagating, cytokine-mediated molecular and cellular reactions. As studies regarding inflammatory changes in cerebrospinal fluid of patients with dementia of the Alzheimer type has been inconclusive, we set up a prospective study to assess cerebrospinal fluid levels of interleukin-1β, interleukin-6, interleukin-10, interleukin-12, soluble interleukin-2 receptor, interferon-γ, tumor necrosis factor-α and neopterin in 20 patients with dementia of the Alzheimer type and 20 age- and sex-matched controls. Comparing both groups, no significant differences in concentrations and specific activities could be revealed. An additional 22 patients were included to enlarge the study population. No statistically significant differences were shown comparing patients (n=42) with the control group (n=20). We conclude that the immune-mediated inflammatory changes found in histopathological studies are not reflected in cerebrospinal fluid of patients with dementia of the Alzheimer type. Probably, cytokine production appears very localized in the central nervous system, not allowing representative detection in cerebrospinal fluid.Further studies assessing cytokine levels in various regions of central nervous system of patients with dementia of the Alzheimer type will be of interest to confirm this hypothesis.     

Declining incidence of AIDS dementia complex after introduction of zidovudine treatment.

OBJECTIVE--To assess the incidence of the AIDS dementia complex and the presence of HIV I p24 antigen in cerebrospinal fluid in relation to zidovudine treatment. DESIGN--Retrospective study of a consecutive series of patients with AIDS from 1982 to 1988. SETTING--An academic centre for AIDS. PATIENTS--196 Patients with AIDS and neurological symptoms examined from 1982 to 1988. INTERVENTIONS--Zidovudine treatment, which was introduced to The Netherlands on 1 May 1987 for patients with severe symptoms of HIV infection (Centers for Disease Control groups IVA, B, C, and D). MAIN OUTCOME MEASURES--Diagnosis of AIDS dementia complex and presence of HIV I p24 antigen in cerebrospinal fluid. RESULTS--The AIDS dementia complex was diagnosed in 40 of the 196 (20%) patients with AIDS. Thirty eight of 107 patients with AIDS (36%) not taking zidovudine developed the AIDS dementia complex compared with two of the 89 (2%) taking the drug (p less than 0.00001). The incidence of the AIDS dementia complex increased to 53% in the first half of 1987, after the introduction of zidovudine in May 1987, decreasing to 10% in the second half of 1987 and to 3% in 1988. Dementia was diagnosed before definition of the AIDS dementia complex (1986) according to DSM-III criteria and there was good agreement between diagnosis before and after 1986. Sixteen of 61 samples of cerebrospinal fluid (26%) from patients with AIDS (10 with the AIDS dementia complex) not taking zidovudine were positive for HIV I p24 antigen, whereas none of 37 cerebrospinal fluid samples from patients with AIDS (two with the AIDS dementia complex) taking zidovudine were positive. CONCLUSIONS--The incidence of AIDS dementia complex in patients with AIDS declined after the introduction of systematic treatment with zidovudine; the AIDS dementia complex might be prevented by inhibiting viral replication in the central nervous system.     

Changes in lectin binding patterns of Leydig cells during fetal and postnatal development in mice

Summary Changes in the lectin binding of mouse Leydig cells during fetal and postnatal development were examined by light- and electron-microscopy using eight different biotinylated lectins (ConA, WGA, RCA-I, UEA-I, GS-I, PNA, SBA and GS-II). At the light-microscopic level, ConA, WGA, RCA-I, UEA-I and GS-I showed the same binding pattern in which all five lectins bound to the plasma membrane and cytoplasm of Leydig cells from the 13th day post coitum (p.c.) to the 8th postnatal week. PNA, SBA and GS-II reactions were positive in the plasma membrane and cytoplasm of Leydig cells from the 13th day p.c. to 15th day post partum (p.p.) but disappeared completely by day 20. At the electron-microscopic level, gold particles representing the GS-I or GS-II binding sites were distributed primarily along the cell surface membrane, including that of microvilli, as well as in the cytoplasm. These results indicate that certain glycoconjugates bearingD-galactose,N-acetyl-D-galactosamine, andN-acetyl-D-glucosamine residues are expressed on the cell surface and in the cytoplasm of Leydig cells during the period from the 13th day p.c. to around the 20th day p.p. The results suggest that these glycoconjugates might play some role in modulating hormone-receptor interaction in the Leydig cells before the 20th day. Furthermore, these results may indicate that sugar residues expressed on the cell surface and in the cytoplasm of Leydig cells are different from those in the fetal-neonatal and adult phases.     

Impact of beta-glucan on the faecal microbiota of polypectomized patients: a pilot study

Beta-glucans are polysaccharides present in the cell walls of higher plants, in the seeds of some cereals, and certain yeasts and fungi also produce them. It is suggested that they exhibit, among many other health benefits, protective effects against carcinogenesis in the colon, but there is not enough human data to support this. The aim of the study was to determine the effect of barley-derived beta-glucan in the gut microbiota of polypectomized patients. Subjects were randomly assigned to consume 125 g of bread per day with beta-glucan (3 g/d), or without (placebo group), for 3 months. Thirty-three polypectomized men and women (mean age 57.6 years) were recruited into the study, but only 20 completed. Subjects did not consume any probiotics, prebiotics or antibiotics 2 months prior the intervention, or during the study. Stool samples were collected at baseline, on days 30 and 90 of intervention, as well as 2 weeks after the intervention, for enumeration of total aerobes and anaerobes, coliforms, E. coli, enterococci, Bacteroides spp., Clostridium perfringens, bifidobacteria, lactobacilli and Candida spp. Faecal bacterial enzyme activity (beta-glucuronidase and beta-glucosidase), pH, faecal moisture and the concentration of volatile fatty acids in the faeces were measured. Gastrointestinal symptoms were also recorded. Overall, no significant differences were observed in bacterial viable counts between the two feeding groups. Group specific analysis for β-glucan group revealed significantly decreased total coliform counts on the 30th day of the trial compared to the baseline (p = 0.041). Clostridium perfringens concentration increased without reaching statistical significance, on the 30th day, while it decreased significantly on the 90th day of the intervention compared to the 30th day (p = 0.016). An increase was noted in the molar ratio of acetate on the 90th day of the trial compared to placebo (p = 0.018). The molar ratio of butyrate presented a trend to increase on the 30th day, which decreased (p = 0.013) on the 90th day and then increase 2 weeks after the intervention (p = 0.017) compared to placebo. A decrease was recorded in the β-glucan group in the bloating and abdominal pain score after the 30th day of the intervention (Day 30–37) compared to placebo. During β-glucan administration we did not observe any changes on beta-glucuronidase or beta-glucosidase activity, faecal pH, or on faecal moisture.     

Isolation of Saccharomyces cerevisiae strains from different food matrices and their preliminary selection for a potential use as probiotics

Aims: To isolate acid‐ and bile‐resistant Saccharomyces cerevisiae strains directly from food samples and to preliminarily select them on the basis of fundamental probiotic properties. Methods and Results: A rapid screening method allowed the isolation and selection of 20 acid‐ and bile‐resistant yeasts from foods, avoiding time‐consuming isolation steps. The strains were characterized for their specific survival in simulated gastric juice and in intestinal fluid after pre‐exposure at low pH. Ten isolates demonstrated a satisfactory survival percentage in intestinal fluid after pre‐exposure to gastric juice and appreciable lipolytic and proteolytic properties, as demonstrated by the API‐ZYM test. By using molecular methods five strains were identified as Saccharomyces cerevisiae, three as Candida spp., one as Candida pararugosa and one as Pichia spp. The Saccharomyces cerevisiae strains showed considerable probiotic properties, achieving a 80< % <90 survival through the simulated gastrointestinal tract, as well as interesting glucosidase activities. Conclusions: The research represents an efficient strategy to select and identify Saccharomyces cerevisiae strains with desirable acid and bile resistances. Significance and Impact of the Study: This paper reports the direct selection of potentially probiotic yeasts from foods and provides indications about the ability of Saccharomyces cerevisiae strains to survive conditions simulating the human gastrointestinal tract.     

Age-dependent responses of Phaseolus aureus Roxb. to inorganic salts of mercury and cadmium

Phaseolus aureus Roxb. was exposed to Hg and Cd separately at different stages/ages of its development, viz. seed germination stage and seedling stages (4th and 6th day). The responses, besides being metal specific, were also age-dependent. The root growth study at germination stage treatment (GST) revealed Hg to be more toxic than Cd, but, in contrast, at seedling stage treatment (SST) with seedlings more than 5 days old, while Cd killed all the seedlings at 30 μM concentration, Hg did not even at 200 μM. Among the enzymes studied, catalase showed greater metal specific and age-dependent responses than the peroxidases. Both the metals significantly increased the levels of chlorophylls and carotenoids at GST (25 μM Hg/Cd) and 4th day SST (20 μM Hg/Cd), but not at 6th day SST (20 μM Hg/Cd). The photosynthetic O₂ evolution rate expressed as per unit chlorophyll (chl) decreased irrespective of the treatment stages, and also the metals; however, when expressed as per unit f. w., it was inhibited only at 6th day SST, exclusively by Cd. It seems that plants, unlike animals, are capable of facing challenges of metals more at younger than at older stages of their development, probably by mechanisms very different from those genetically controlled.