Zinc deficiency leads to reduced interleukin-2 production by active gene silencing due to enhanced CREMα expression in T cells.

BACKGROUND & AIMS: The micronutrient zinc is essential for proper immune function. Consequently, zinc deficiency leads to impaired immune function, as seen in decreased secretion of interleukin (IL)-2 by T cells. Although this association has been known since the late 1980s, the underlying molecular mechanisms are still unknown. Zinc deficiency and reduced IL-2 levels are especially found in the elderly, which in turn are prone to chronic diseases. Here, we describe a new molecular link between zinc deficiency and reduced IL-2 expression in T cells. METHODS: The effects of zinc deficiency were first investigated in vitro in the human T cell lines Jurkat and Hut-78 and complemented by in vivo data from zinc-supplemented pigs. A short- and long-term model for zinc deficiency was established. Zinc levels were detected by flow cytometry and expression profiles were investigated on the mRNA and protein level. RESULTS: The expression of the transcription factor cAMP-responsive-element modulator α (CREMα) is increased during zinc deficiency in vitro, due to increased protein phosphatase 2A (PP2A) activity, resulting in decreased IL-2 production. Additionally, zinc supplementation in vivo reduced CREMα levels causing increased IL-2 expression. On epigenetic levels increased CREMα binding to the IL-2 promoter is mediated by histone deacetylase 1 (HDAC1). The HDAC1 activity is inhibited by zinc. Moreover, deacetylation of the activating histone mark H3K9 was increased under zinc deficiency, resulting in reduced IL-2 expression. CONCLUSIONS: With the transcription factor CREMα a molecular link was uncovered, connecting zinc deficiency with reduced IL-2 production due to enhanced PP2A and HDAC1 activity.

Donor age of human platelet lysate affects proliferation and differentiation of mesenchymal stem cells.

The regenerative potential declines upon aging. This might be due to cell-intrinsic changes in stem and progenitor cells or to influences by the microenvironment. Mesenchymal stem cells (MSC) raise high hopes in regenerative medicine. They are usually culture expanded in media with fetal calf serum (FCS) or other serum supplements such as human platelet lysate (HPL). In this study, we have analyzed the impact of HPL-donor age on culture expansion. 31 single donor derived HPLs (25 to 57 years old) were simultaneously compared for culture of MSC. Proliferation of MSC did not reveal a clear association with platelet counts of HPL donors or growth factors concentrations (PDGF-AB, TGF-ß1, bFGF, or IGF-1), but it was significantly higher with HPLs from younger donors (<35 years) as compared to older donors (>45 years). Furthermore, HPLs from older donors increased activity of senescence-associated beta-galactosidase (SA-ßgal). HPL-donor age did not affect the fibroblastoid colony-forming unit (CFU-f) frequency, immunophenotype or induction of adipogenic differentiation, whereas osteogenic differentiation was significantly lower with HPLs from older donors. Concentrations of various growth factors (PDGF-AB, TGF-ß1, bFGF, IGF-1) or hormones (estradiol, parathormone, leptin, 1,25 vitamin D3) were not associated with HPL-donor age or MSC growth. Taken together, our data support the notion that aging is associated with systemic feedback mechanisms acting on stem and progenitor cells, and this is also relevant for serum supplements in cell culture: HPLs derived from younger donors facilitate enhanced expansion and more pronounced osteogenic differentiation.

Measurement of medial olivocochlear efferent activity in humans: comparison of different distortion product otoacoustic emission-based paradigms.

OBJECTIVE: To assess the suitability of contralateral suppression (CS) of distortion product otoacoustic emissions (DPOAEs) for measurement of activity of the medial olivocochlear (MOC) efferents. BACKGROUND: The MOC efferent system has been shown to be involved in sound discrimination, selective attention to tones, sound localization, and protection of the cochlea against noise. A great variety of paradigms for measurement of MOC activity by CS of OAE (MOC reflex [MOCR]), has been described. An issue of this approach is the dependence of the CS values on stimulus parameters, especially when DPOAE are used. METHODS: Four different measurement paradigms, which used different combinations of stimulus frequencies and primary tone levels, were applied in 16 human subjects. RESULTS: Mean absolute values of CS were in the range of 1.2 to 2.6 dB. The use of different stimulus parameters produced not only MOCR values of different size-which was expected-but, in many cases, also different relative classifications of the subjects according to their MOCR strength. CONCLUSION: The suppression effects on DPOAE demonstrated in this study reflect MOC activity. However, the new conclusion from our data is that CS of DPOAE measurements, as they were used in this study, may not allow for a consistent quantitative classification of human subjects according to their MOCR strength. This finding concerns interpretation of previous studies using CS of DPOAE and analogous future studies. One future approach may lie in the separation of the DPOAE components to distinguish interference phenomena, which complicate interpretation of CS values.

Endoscopic transventricular third ventriculostomy through the lamina terminalis.

OBJECT: Endoscopic third ventriculostomy (ETV) has become a well-accepted option for obstructive hydrocephalus. However, standard ventriculostomy at the floor of the third ventricle might not be feasible under certain conditions. Here, the authors report in detail on their initial experience with an alternative option of endoscopic ventriculostomy through the lamina terminalis via a transventricular route. METHODS: Endoscopic third ventriculostomy through the lamina terminalis from a transventricular transforaminal route was evaluated in 4 cadaveric human heads and in 4 clinical cases. RESULTS: In all 4 human cadavers, an opening of the lamina terminalis via a transventricular approach could be achieved without injury to either the optic chiasm or the anterior cerebral arteries. In the 4 clinical cases, an accurate and reliable ventriculostomy was performed at the lamina terminalis. The bur hole was placed directly at the coronal suture 2 cm lateral from the midline. After identifying the optic chiasm and the anterior cerebral arteries, a blunt perforation was made just anterior to the optic chiasm by using perforation forceps and a balloon catheter. After the opening, the stoma was inspected with a 0° and 30° rod lens endoscope, and its patency as well as the preservation of vessels and optic nerves was checked. No complications occurred, although all patients suffered from a clinically silent fornical contusion at the foramen of Monro. CONCLUSIONS: Endoscopic opening of the lamina terminalis via a transventricular transforaminal route appears to be feasible. No complications were observed. Although no conclusions on the clinical success rate can be drawn, the reliable anatomical opening and known success rate for anterior subfrontal approaches suggest that the technique represents an alternative in a small subgroup of patients in whom a standard ETV cannot be performed.

Resistance of polysaccharide coatings to proteins, hematopoietic cells, and marine organisms.

The interaction of covalently coupled hyaluronic acid, alginic acid, and pectic acid with proteins, cells (hematopoietic KG1a and Jurkat cells), and marine organisms (algal zoospores and barnacle cypris larvae) is compared. In contrast to cells and proteins for which such polysaccharide coatings are known for their antiadhesive properties, marine algal spores and barnacle cyprids were able to colonize the surfaces. Of the three polysaccharides, hyaluronic acid showed the lowest settlement of both Ulva zoopores and barnacles. Photoelectron spectroscopy reveals that the polysaccharide coatings tend to bind bivalent ions, such as calcium, from salt water. Such pretreatment with a high salinity medium significantly changes the protein and hematopoietic cell resistance of the surfaces. Complexation of bivalent ions is therefore considered as one reason for the decreased resistance of polysaccharide coatings when applied in the marine environment.

Test-retest repeatability of distortion product otoacoustic emissions.

OBJECTIVES: Distortion product otoacoustic emissions (DPOAE) have become part of routine audiological diagnostics. The large scale of clinical DPOAE applications, such as screening of hearing in infants, objective estimation of hearing status, distinction between cochlear and retrocochlear origin of sensorineural hearing loss, exclusion of psychogenic hearing loss, monitoring of hearing during administration of ototoxic drugs, and others illustrates the significance of this audiological tool. In all diagnostic tests, knowledge about the procedure's test-retest repeatability is of crucial importance, to allow for distinction between measurement deviations and true physiological or pathological changes in monitoring over time. DESIGN: Measurements of DPOAE were performed in triplicate in 80 normally hearing ears of 40 subjects. Both immediate remeasurements with the ear probe left in place [single-fit mode (SF-mode)] and remeasurements after approximately 5 to 10 days [multiple-fit mode (MF-mode)] were included. DPOAE primary tone levels were varied in 5 dB steps from L2 = 60 to 20 dB SPL (L1 = L2 x 0.4 + 39 dB SPL) and within the frequency range f2 = 1 to 6 kHz. Repeatability of DPOAE was evaluated by the standard error of measurement (Sm), reliability (Cronbach alpha), absolute differences between measurements, 95% confidence intervals, and repeatability standard deviations. RESULTS: Sm averaged 0.67 dB over all frequencies and primary tone levels in the SF-mode, and 1.44 dB in the MF-mode, respectively. As expected, test-retest repeatability declined with decreasing primary tone levels; however, repeatability values were still mostly satisfactory with the lower primary tone levels. For the exemplary primary tone level combination of L1/L2 = 63/60 dB SPL, which is close to common clinical paradigms, the difference between two DPOAE measurements under the reported test conditions could be considered statistically significant (p = 0.05) if it exceeded 0.7 to 1.3 dB in the range 1 to 5 kHz and 2.3 dB for 6 kHz in the SF-mode, when compared with 1.8 to 2.7 dB for 1 to 5 kHz and 3.7 dB for 6 kHz in the MF-mode. Signal to noise ratio (SNR) did not seem to have a large influence on repeatability, as long as SNR was within 6 to 35 dB, which covers the range of most clinical DPOAE measurements. CONCLUSIONS: The DPOAE-test-retest study presented here is to our knowledge the first, which combines variation of primary tone levels, assessment of both SF- and MF-modes, and comparison of the two modalities within the same subjects. Although the measurements were conducted under practical conditions resembling the clinical setting, repeatability was generally good. The widely used minimum SNR of 6 dB seems to be a recommendable criterion when considering both practicability and measurement quality under clinical conditions. The current findings underline the suitability of DPOAE as a monitoring tool of cochlear status over time. The data are intended to assist the clinician and the scientist in the correct interpretation of DPOAE level changes in the test-retest situation.

Evidence for a bipolar change in distortion product otoacoustic emissions during contralateral acoustic stimulation in humans.

The aim of this study was to investigate the activity of the medial olivocochlear (MOC) efferents during contralateral (CAS) and ipsilateral acoustic stimulation (IAS) by recording distortion product otoacoustic emission (DPOAE) suppression and DPOAE adaptation in humans. The main question was: do large bipolar changes in DPOAE level (transition from enhancement to suppression) also occur in humans when changing the primary tone level within a small range as described by Maison and Liberman for guinea pigs [J. Neurosci. 20, 4701-4707 (2000)]? In the present study, large bipolar changes in DPOAE level (14 dB on average across subjects) were found during CAS predominantly at frequencies where dips in the DPOAE fine structure occurred. Thus, effects of the second DPOAE source might be responsible for the observed bipolar effect. In contrast, comparable effects were not found during IAS as was reported in guinea pigs. Reproducibility of CAS DPOAEs was better than that for IAS DPOAEs. Thus, contralateral DPOAE suppression is suggested to be superior to ipsilateral DPOAE adaptation with regard to measuring the MOC reflex strength and for evaluating the vulnerability of the cochlea to acoustic overexposure in a clinical context.