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PubMed1990

Optimum allocation of samples in strata-matching case-control studies when cost per sample differs from stratum to stratum.

We investigate the allocation of samples over strata in case-control studies with consideration of sampling costs. We consider two situations, typical of strata-matched case-control studies. Subject to a fixed total cost we derived optimum allocations that maximize the asymptotic efficiency of Cochran's test, that is, minimize the variance of the maximum likelihood estimator of the common odds ratio when the common odds ratio is one. Our results show that the standard design with equal numbers of cases and controls in each stratum is inefficient when costs per control differ greatly from stratum to stratum. We show that the optimum design suggested in this paper is robust for values of the odds ratio common in epidemiologic studies.

Case-Control Studies↗
PubMed2000

The activity of fatty acid synthase of epidermal keratinocytes is regulated in the lower stratum spinousum and the stratum basale by local inflammation rather than by circulating hormones.

The epidermal keratinocytes produce and secrete lipids to maintain the water barrier of the epidermis. To clarify the regulation of epidermal lipid synthesis, we investigated the hormonal effect on the activity of fatty acid synthase (FAS) of the keratinocytes, and the expression of FAS in the human skin. In cultured keratinocytes, the FAS activity, assayed by measuring the oxidation of NADPH, was slightly increased by hydrocortisone or testosterone, but not influenced by thyroid hormone, estrogen, progesterone or insulin. In immunohistochemical study of normal human epidermis, FAS was expressed strongly in the stratum granulosum and moderately in the uppermost layer of the stratum spinousum (SS), suggesting that fatty acid synthesis may increase during normal epidermal differentiation. In inflammatory disorders, such as psoriasis, lichen planus, and atopic dermatitis, FAS was also expressed in the lower SS and the stratum basale (SB), resulting in strong staining in the whole layers of the epidermis. Remarkable increase of FAS expression was only observed in the lower SS and the SB. Therefore, the activity of FAS in the epidermis may be regulated in the lower SS and the SB by local inflammation rather than by circulating hormones. In other components of the skin, FAS was strongly expressed not only in adipose tissue and sebaceous glands, which are known as active sites of lipid synthesis, but also in sweat glands, suggesting that the sweat glands can synthesize abundant fatty acids de novo.

Antibodies↗
PubMed1994

Levels of terbinafine in plasma, stratum corneum, dermis-epidermis (without stratum corneum), sebum, hair and nails during and after 250 mg terbinafine orally once daily for 7 and 14 days.

In earlier skin pharmacokinetic studies we have shown that terbinafine is rapidly delivered to the stratum corneum, nails and hair both through sebum and by direct diffusion through dermis-epidermis. In the present study the skin pharmacokinetic profile of terbinafine was studied in two groups of eight human male volunteers during and after 250 mg orally once daily for 7 and 14 days. In the 7-day study high terbinafine levels were found in sebum (19.0 micrograms/g) and stratum corneum (2.5 micrograms/g), and a concentration in stratum corneum above the minimal inhibitory concentration for most dermatophytes was still found 48 days after the last day of medication. Terbinafine was found in peripheral nail clippings after 7 days of medication and the concentration was, in the 7-day study, 0.5 microgram/g 1 day after stopping medication; it was still 0.2 microgram/g 90 days after stopping treatment. The results in the 14-day study were in parallel with, but higher than, in the 7-day study. The elimination of terbinafine from several compartments is biphasic, with a faster initial elimination followed by a slower secondary elimination. For nails, the elimination is slower compared with the other compartments. The results indicate that terbinafine may be effective in short-term treatment of several dermatophytoses. The concentration of 0.2 microgram/g of terbinafine found in nails 90 days after stopping medication, following 7 days of treatment, indicates that the duration of therapy, even in tinea ungium, may be shorter than is currently the case.

Administration, Oral↗
PubMed1992

A distinct stratum corneum antigen in psoriasis and its reactions with stratum corneum autoantibodies.

Stratum corneum antibodies are ubiquitous and can be detected by various immunological methods. Of these, the ones detected by hemagglutination undergo changes in antibody titers and have been implicated in psoriasis. The purpose of our study was to examine if differences exist in the activities of the antigens isolated from psoriatic scales in comparison to normal callus. Stratum corneum antigens were prepared by trypsin-phenol-water extraction. The water phase, which contains the stratum corneum antigen, was used to sensitize the red blood cells in the hemagglutination assay. The antibody activity in human sera was determined before and after absorption with antigens isolated from callus, psoriatic scales, and cell envelopes. We found notable differences in the antigens obtained from callus and psoriatic scales. These include higher antibody titers to the antigens of the scales, the presence of unique antigenic determinants on psoriatic scales and the localization of the antigen on cell envelopes. These immunological differences were corroborated by the marked biochemical differences of certain amino acids, most notably glycine and proline, and these differences were unique to psoriatic scales as they were not shared with other hyperproliferative disorders.

Amino Acids↗
PubMed1997

Interneurons in area CA1 stratum radiatum and stratum oriens remain functionally connected to excitatory synaptic input in chronically epileptic animals.

Past work has demonstrated a reduction of stimulus-evoked inhibitory input to hippocampal CA1 pyramidal cells in chronic models of temporal lobe epilepsy (TLE). It has been postulated that this reduction in inhibition results from impaired excitation of inhibitory interneurons. In this report, we evaluate the connectivity of area CA1 interneurons to their excitatory afferents in hippocampal-parahippocampal slices obtained from a rat model of chronic TLE. Rats were made chronically epileptic by a period of continuous electrical stimulation of the hippocampus, which establishes an acute condition of self-sustained limbic status epilepticus (SSLSE). This period of SSLSE is followed by a development of chronic recurrent spontaneous limbic seizures that are associated with chronic neuropathological changes reminiscent of those encountered in human TLE. Under visual control, whole cell patch-clamp recordings of interneurons and pyramidal cells were obtained in area CA1 of slices taken from adult, chronically epileptic post-SSLSE rats. Neurons were activated by means of electrodes positioned in stratum radiatum. Intrinsic membrane properties, including resting membrane potential, action potential (AP) threshold, AP half-height width, and membrane impedance, were unchanged in interneurons from chronically epileptic (post-SSLSE) tissue compared with control tissue. Single stimuli delivered to stratum radiatum evoked depolarizing excitatory postsynaptic potentials and APs in interneurons, whereas paired-pulse stimulation evoked facilitation of the postsynaptic current (PSC) in both control and post-SSLSE tissue. No differences between interneurons in control versus post-SSLSE tissue could be found with respect to the mean stimulus intensity or mean stimulus duration needed to evoke an AP. A multiple linear regression analysis over a range of stimulus intensities demonstrated that a greater number of APs could be evoked in interneurons in post-SSLSE tissue compared with control tissue. Spontaneous PSCs were observed in area CA1 interneurons in both control and post-SSLSE tissue and were markedly attenuated by glutamatergic antagonists. In conclusion, our data suggest that stimulus-evoked and spontaneous excitatory synaptic input to area CA1 interneurons remains functional in an animal model of chronic temporal lobe epilepsy. These findings suggest, therefore, that the apparent decrease of polysynaptic inhibitory PSPs in CA1 pyramidal cells in epileptic tissue is not due to a deficit in excitatory transmission from Schaffer collaterals to interneurons in stratum radiatum and straum oriens.

Animals↗
PubMed1990

Pertussis toxin prevents induction of hippocampal long-term potentiation in the stratum radiatum and stratum oriens inputs to CA1 neurons.

Stereotaxic injections of pertussis toxin (3-4 micrograms) over the right hippocampus resulted in blockade of long-term potentiation (LTP) induction in the ipsilateral stratum radiatum-CA1 and stratum oriens-CA1 synaptic systems. LTP of intracellularly recorded excitatory postsynaptic potentials was prevented in slices obtained from the hippocampus at 3 and 4 but not 6 days post-toxin injection. Slices taken from the left (contralateral) hippocampus on the same days as above exhibited LTP which was similar to that obtained in control slices from uninjected rats. The post- but not presynaptic actions of adenosine were antagonized at 3, 4 and 6 days post-toxin injection. The observations suggest that the guanosine triphosphate binding proteins involved in LTP induction (GLTP) and those coupled to the postsynaptic adenosine receptors exhibit different turnover times.

Action Potentials↗
PubMed1994

Loss of calbindin-immunoreactivity in CA1 hippocampal stratum radiatum and stratum lacunosum-moleculare interneurons in the aged rat.

Alterations in hippocampal circuitry may underly age-related learning and memory impairment. We showed in a previous study that the GABAB-mediated slow inhibitory postsynaptic potential (IPSP) induced in CA1 pyramidal neurons by electrical stimulation of stratum radiatum, is depressed in the hippocampus of the aged rat. This could be due to alterations in GABAergic interneuron functions. We report in this study that the number of hippocampal calbindin-immunoreactive (CaBP-IR) GABAergic interneurons is decreased in the aged rat. The mean number of CaBP-IR interneurons per slice decreases by 50% in the aged rat. The most severe loss was observed in the stratum radiatum of CA1 (78%), with a less consistent loss of immunoreactivity in CA3 (35%). In contrast, the mean number of interneurons containing parvalbumin (PV), was not significantly decreased in the aged rat. Our results show a loss of CaBP immunoreactivity in a population of GABAergic interneurons, which might be related to an altered function of these interneurons and consequently of GABAergic synaptic transmission in the aged rat. In contrast, PV immunoreactivity in interneurons located close to the pyramidal layer does not decrease in the hippocampus of the aged rat.

Aging↗
PubMed1975

The mechanical properties of stratum corneum. I. The effect of water and ambient temperature on the tensile properties of newborn rat stratum corneum.

The tensile properties of the outermost layer of skin of neonatal rats, the stratum corneum, were investigated at a constant strain rate as a function of moisture content and ambient test temperature. The results show that the mechnical behavior of this membrane, whose primary constituent is the fibrous protein keratin, can be significantly altered by variations in both the sorbed water content and ambient temperature. In particular, a brittle to ductile transition was observed at 25 degrees C once the hydration level exceeded 70% relative humidity. Similarly, an identical phenomenon was detected at temperatures beyond 40 degrees C for specimens whose equilibrium moisture concentrations were maintained at 10 g H2 O/100 g dry protein. Differential scanning calrimetry measurements showed the presence of a molecular relaxation process which migrated from 42 degrees C at 40% relative humidity to --18 degrees C at 95% relative humidity. It is postulated that this relaxation process, possibly corresponding to the glass transition of the fibrous protein component of stratum corneum, is primarily responsible for the observed behavior.

Animals↗
PubMed2003

Different patterns of synaptic transmission revealed between hippocampal CA3 stratum oriens and stratum lucidum interneurons and their pyramidal cell targets.

Stratum lucidum (SL) interneurons likely mediate feedforward inhibition between the dentate gyrus mossy fibers and CA3 pyramidal cells, while stratum oriens (SO) interneurons likely provide both feedforward and feedback inhibition within the CA3 commissural/associational network. Using dual whole-cell patch-clamp recordings between interneurons and CA3 pyramidal cells, we have examined SL and SO interneurons and their synapses within organotypic hippocampal slice cultures. Biocytin staining revealed different morphologies between these interneuron groups, both being very similar to those found previously in acute slices. The kinetics of IPSCs were similar between the two groups, but the reliability of synaptic transmission of SL interneuron (SL-INT) IPSCs was significantly lower than the virtually 100% reliability (non-existent failure rates) of SO-INT IPSCs. The SL-INT IPSCs also had a lower quantal content than the SO-INT IPSCs. In addition, SL-INTs were less likely than SO-INTs to innervate or to be innervated by nearby CA3 pyramidal cells. Paired-pulse stimulation at 100 ms interstimulus intervals produced similar paired-pulse depression in both interneuron synapses, despite the significantly higher failure rate of IPSCs produced by the SL-INTs compared with SO-INTs. CV analysis supported the hypothesis that paired-pulse depression was presynaptic. During repetitive, high frequency stimulation (>10 Hz for 500 ms) the two different synapses exhibited distinctly different forms of short-term plasticity: all SL interneurons displayed significant short-term facilitation (mean 113% facilitation, n=4), while, by contrast, SO interneuron synapses displayed either short-term depression (mean 42% depression, n=5 of 8) or no net facilitation or depression (n=3 of 8). These results indicate that the synaptic properties of interneurons can be quite different for interneurons in different hippocampal circuits.

Action Potentials↗
PubMed2004

Stratum corneum acidification in neonatal skin: secretory phospholipase A2 and the sodium/hydrogen antiporter-1 acidify neonatal rat stratum corneum.

At birth, human stratum corneum (SC) displays a near-neutral surface pH, which declines over several days to weeks to months to an acidic pH, comparable to that of adults. Recent studies suggest that an acidic pH is required for normal permeability barrier homeostasis and SC integrity/cohesion. We assessed here the basis for postnatal acidification in the neonatal rat, where SC pH, as measured with a flat surface electrode, declines progressively from near-neutral levels (pH 6.63) on postnatal days 0 to 1 to adult levels (pH 5.9) or even below over the subsequent 7 to 8 d. The postnatal decline in SC pH was paralleled by a progressive activation of a pH-dependent hydrolytic enzyme, beta-glucocerebrosidase. Because SC acidification could not be linked to commonly implicated exogenous factors, such as bacterial colonization, or the deposition of sebaceous gland products. We next assessed whether changes in one or more of three endogenous mechanisms demonstrate postnatal activity changes that contribute to the progressive development of an acidic SC pH. Although the histidine-to-urocanic acid pathway has been implicated in acidification of the adult SC, surface pH is completely normal in histidase-deficient (his/his, Peruvian) mice, ruling out a requirement for this mechanism. In contrast, when sodium/hydrogen antiporter-1 (NHE1), which predominantly acidifies membrane domains at the stratum granulosum-SC interface, is inhibited, postnatal acidification of the SC is partially blocked. Likewise, SC secretory phospholipase A2 (sPLA2) activity, measured with a fluorometric assay, is low at birth, but increases progressively (by 66%) over the first 5 d after birth, and inhibition of sPLA2 between days 0 to 1 and days 5 to 6 delays postnatal SC acidification. Together, these results describe a neonatal model, in which the development of an acidic surface pH can be ascribed, in part, to progressive SC acidification by two endogenous mechanisms, namely, sPLA2 and NHE1, which are known to be important for acidification of adult rodent SC. Conversely, the impaired acidification of neonatal SC, which has important functional and clinical consequences, can be explained by the relatively low activities of one or both of these mechanisms at birth.

Acids↗
PubMed2004

Stratum-by-stratum projection of light response attributes by retinal bipolar cells of Ambystoma.

The visual system processes light images by projecting various representations of the visual world to segregated regions in the brain through parallel channels. Retinal bipolar cells constitute the first parallel channels that carry different light response attributes to different parts of the inner plexiform layer (IPL). Here we present a systematic study on detailed axonal morphology and light response characteristics of over 200 bipolar cells in dark-adapted salamander retinal slices by the whole-cell voltage clamp and Lucifer yellow fluorescence (with a confocal microscope) techniques. Four major groups of bipolar cells were identified according to the patterns of axon terminal ramification in the IPL: 36% were narrowly monostratified (whose axon terminals ramified in one of the 10 strata of the IPL), 27% were broadly monostratified, 19% were multistratified, and 18% bore pyramidally branching axons. By analysing the bipolar cells with narrowly monostratified axon terminals in each of the 10 strata of the IPL, we found that several key light response attributes are highly correlated with the strata in which the cells' axon terminals ramify. The 10 IPL strata appear to be the basic building blocks for attributes of light-evoked signal outputs in all bipolar cells, and several general stratum-by-stratum rules were identified by analysing the broadly monostratified, multistratified and pyramidally branching cells. These rules not only uncover mechanisms by which third-order retinal cells integrate and compute bipolar cell signals, but also shed considerable light on how bipolar cells in other vertebrates process visual information and how physiological signals may shape the morphology and projection of output synapses of visual neurones during development.

Adaptation, Ocular↗
PubMed2000

Comparison of changes evoked by GABA (gamma-aminobutyric acid) and anoxia in [K+]o, [Cl-]o, and [Na+]o in stratum pyramidale and stratum radiatum of the guinea pig hippocampus.

Ion-selective microelectrode recordings were made to assess a possible contribution of extracellular gamma-aminobutyric acid (GABA) accumulation to early responses evoked in the brain by anoxia and ischemia. Changes evoked by GABA or N2 in [K+]o, [Cl-]o, [Na+]o, and [TMA+]o were recorded in the cell body and dendritic regions of the stratum pyramidale (SP) and stratum radiatum (SR), respectively, of pyramidal neurons in CA1 of guinea pig hippocampal slices. Bath application of GABA (1-10 mM) for approximately 5 min evoked changes in [K+]o and [Cl-]o with respective EC50 levels of 3.8 and 4.1 mM in SP, and 4.7 and 5.6 mM in SR. In SP 5 mM GABA reversibly increased [K+]o and [Cl-]o and decreased [Na+]o; replacement of 95% O2 -5% CO2 by 95% N2 -5% CO2 for a similar period of time evoked changes which were for each ion in the same direction as those with GABA. In SR both GABA and N2 caused increases in [K+]o and decreases in [Cl-]o and [Na+]. The reduction of extracellular space, estimated from levels of [TMA+]o during exposures to GABA and N2, was 5-6% and insufficient to cause the observed changes in ion concentration. Ion changes induced by GABA and N2 were reversibly attenuated by the GABA(A) receptor antagonist bicuculline methiodide (BMI, 100 microM). GABA-evoked changes in [K+]o in SP and SR and [Cl-]o in SP were depressed by > or =90%, and of [Cl-]o in SR by 50%; N2-evoked changes in [K+]o in SP and SR were decreased by 70% and those of [Cl-]o by 50%. BMI blocked delta [Na+]o with both GABA and N2 by 20-30%. It is concluded that during early anoxia: (i) accumulation of GABA and activation of GABA(A) receptors may contribute to the ion changes and play a significant role, and (ii) responses in the dendritic (SR) regions are greater than and (or) differ from those in the somal (SP) layers. A large component of the [K+]o increase may involve a GABA-evoked Ca2+-activated gk, secondary to [Ca2+]i increase. A major part of [Cl-]o changes may arise from GABA-induced g(Cl) and glial efflux, with strong stimulation of active outward transport and anion exchange at SP, and inward Na+/K+/2Cl- co-transport at SR. Na+ influx is attributable mainly to Na+-dependent transmitter uptake, with only a small amount related to GABA(A) receptor activation. Although the release and (or) accumulation of GABA during anoxia might be viewed as potentially protectant, the ultimate role may more likely be an important contribution to toxicity and delayed neuronal death.

Animals↗
PubMed2004

Theta rhythmic stimulation of stratum lacunosum-moleculare in rat hippocampus contributes to associative LTP at a phase offset in stratum radiatum.

Computational modeling demonstrates that encoding and context-dependent retrieval of memories in region CA1 of the hippocampus will be most effective when the phase of strongest entorhinal input (to stratum lacunosum-moleculare) is offset from the phase of maximal induction of long-term potentiation at Schaffer collateral synapses (in s. radiatum). This would allow entorhinal input to play a role in both retrieval and encoding without engaging long-term potentiation (LTP) during retrieval. Experiments in brain slice preparations of the hippocampal formation tested the relationship between rhythmic input to s. lacunosum-moleculare and the time of maximal LTP induction at Schaffer collateral synapses in s. radiatum. Analysis of the data demonstrates a statistically significant difference in the induction of LTP for different time intervals between the end of each four-pulse train in s. lacunosum-moleculare and the single pulse s. radiatum stimulation. The time of maximal LTP induction was found to be approximately 30 ms after the end of lacunosum-moleculare stimulation, consistent with the requirements of the model.

Animals↗
PubMedDate not supplied

Reservoir function of the stratum corneum: development of an in vivo method to quantitatively determine the stratum corneum reservoir for topically applied substances.

Investigations on the stratum corneum (SC) reservoir for topically applied substances are of importance in dermatologic science in order to assess the pharmacokinetics of these substances. In the present study, an in vivo method was developed to determine the SC reservoir quantitatively and to investigate the temporal behavior of this reservoir. Therefore, increasing amounts of an oil-in-water emulsion (o/w emulsion) containing 4% of a chemical UV filter were topically applied onto the flexor forearms of 5 healthy volunteers. The saturation of the SC reservoir was determined utilizing the tape stripping technique 1 and 6 h after application. The capacity of the SC reservoir for the o/w emulsion was found to be approximately 2.7 mg/cm(2). Furthermore, a correlation of the capacity of the SC with transepidermal water loss was observed. Extending the time between the topical application and SC removal did not affect the distribution or the recovery rate of the UV filter in the SC. The results indicate that the reservoir of the SC is limited. This is reflected by the saturation level, which depends on the individual volunteer and, presumably, the topically applied substances and formulations used. The results show that the method developed is suited to quantitatively determine in vivo the SC reservoir for topically applied substances.

Administration, Topical↗
PubMed1981

Stratum corneum lipids in disorders of cornification: increased cholesterol sulfate content of stratum corneum in recessive x-linked ichthyosis.

Activity of the microsomal enzyme, steroid sulfatase, is absent in keratinocytes, fibroblasts, and leukocytes of patients with recessive x-linked ichthyosis. This study was undertaken to determine if cholesterol sulfate, a substrate of this enzyme, accumulates in the pathological scale of these patients. Scales from 8 patients with recessive x-linked ichthyosis, 10 patients with other forms of ichthyosis, and normal human outer stratum corneum were extracted with chloroform/water (1:2:0.8 by vol) and lipids were fractionated by quantitative, sequential thin-layer chromatography. Cholesterol sulfate was identified by cochromatography in several solvent systems, by its staining characteristics, by biochemical analysis, and by mass spectrometry. The mean cholesterol sulfate content of recessive x-linked ichthyotic scale was 12.5 +/- 0.8% of the total lipid, a fivefold increase over normal (P less than 0.0025), whereas the cholesterol sulfate content of other ichthyotic scale was normal. This increase in cholesterol sulfate content was accompanied by a decrease in total neutral lipids (P less than 0.0025) and free sterols (P less than 0.025) but no change in sterol esters or total sterols. These results demonstrate that deficiency of steroid sulfatase in recessive x-linked ichthyosis results in excessive accumulation of a substrate, cholesterol sulfate, in the pathologic scale, which may underly the pathogenesis of the scaling in this disorder. Measurement of cholesterol sulfate content in scale provides an alternative method to enzymatic assay for the diagnosis of this form of ichthyosis.

Adolescent↗
PubMed2001

Comparison of bending modulus and yield strength between outer stratum medium and stratum medium zona alba in equine hooves.

OBJECTIVE: To determine whether the bending modulus and yield strength of the outer stratum medium (SM) differed from those of the SM zona alba (SMZA) and to what degree they differed. In addition, a comparison was made among our values and values reported elsewhere. SAMPLE POPULATION: 10 normal equine feet. PROCEDURE: A 3-point bending technique was used to determine the bending modulus and yield strength of the outer SM and SMZA. Efforts were made to minimize biological and technical factors that could influence the bending modulus. RESULTS: Bending modulus of the outer SM was (mean +/- SD) 187.6 +/- 41.3 MPa, whereas mean value for the SMZA was 98.2 +/- 36.8 MPa. Mean yield strength was 19.4 +/- 2.6 MPa for the outer SM and 5.6 +/- 1.7 MPa for the SMZA. Values for bending modulus and yield strength differed significantly between the outer SM and SMZA. Significant differences were not detected when the outer SM was loaded in bending from the outer or inner surface. CONCLUSIONS AND CLINICAL RELEVANCE: Potentially, the SMZA could serve as a mechanical buffer zone between the rigid hoof wall and bone and laminar tissues. This buffer zone potentially assists the feet of horses in transmitting a load through the tissues and prevents the most susceptible tissues from becoming damaged. More consistency among tissue selection, preparation, and testing protocols must be attained before an accurate 3-dimensional finite-element model of an equine foot can be constructed.

Animals↗