Pilot study of cerebral and somatic autoregulation using NIRS in preterm neonates.

BACKGROUND: As neonates transition from a relatively hypoxic environment to extra-uterine life, arterial oxygen saturation dramatically increases. This transition occurs while most organs have not fully matured. The ability for immature tissue to adequately extract and utilize oxygen remains largely unknown. With the development of near-infrared spectroscopy (NIRS), measuring specific tissue oxygen saturation (StO2) noninvasively, clinicians can measure StO2 and determine if adequate tissue oxygenation is maintained. The objective of this study is to determine the relationships of NIRS brain and somatic autoregulation function to patients' severity of illness. METHODS: In this prospective cohort pilot study, after parental consent, neonates less than 34 weeks with arterial access, were enrolled. The FORE-SIGHT NIRS probe was placed on the forehead and abdominal wall for 24 hours. Continuous arterial blood pressure, SpO2 and cerebral and somatic NIRS were used to derive autoregulation function. RESULTS: Data was obtained from 17 neonates (0.540 to 2.37 kg, gestation 23.0 to 33.2 weeks). The autoregulation function categorizes pressure passive index (PPI) values as good, borderline, or poor. For normal autoregulation function, PPI values tend to be low and fairly constant for a range of MAP. The PPI borderline zone is a hypothetical range of PPI values where autoregulation function transitions from good to poor. CONCLUSION: Our results show most premature neonates, as long as they maintained normal BP and systemic circulation can autoregulate cerebral perfusion. When BP are above or below the normal MAP for age, the neonate is at risk for losing brain and somatic autoregulation.

Aqueous solution behaviour and solubilisation properties of octadecyl cationic gemini surfactants and their comparison with their amide gemini analogues.

Gemini surfactants 18-s-18(Et), comprised of two ethylammonium headgroups and two alkyl tails with m = 18 carbon atoms with spacers of s = 4, 6, 8 and 10 linking the headgroups (alkanediyl-α,ω-bis(diethyloctadecylammonium bromides)), were obtained. Their aqueous solution behaviour, including adsorption at the interface and aggregation in solution, was followed by tensiometric, conductometric and spectroscopic methods. The critical micelle concentration (CMC) of the surfactants decreased with increasing spacer length. The size of 18-s-18(Et) aggregates formed at concentrations of 10 and 40 CMC measured by DLS varied with the elongation of the spacer. Visualisation of aggregated surfactant structures at 40 CMC by cryo-TEM evidenced the formation of different morphologies depending on spacer length. Gemini with s = 4 formed elongated, cylindrical micelles, while geminis of s = 6, 8 and 10 self-assembled into vesicles. The ability of the studied geminis to solubilise hydrophobic dye Sudan I in water was determined as a function of surfactant concentration, demonstrating their high efficiency. Results for 18-s-18(Et) geminis were compared with those previously obtained for their analogues containing an amide group placed between headgroups and tails. The significant impact of amide groups on the surface activity and aggregation properties of gemini surfactants was evidenced and is related to hydrogen-bond formation by amide-containing compounds.

Lateral cephalometric standards of Germans with normal occlusion from 6 to 17 years of age.

The aim of this study was to analyze sagittal and vertical dentofacial dimensions in subjects with normal occlusions during the juvenile and adolescents age periods to establish age- and gender-specific lateral cephalometric standard values for Germans during their active growth period. The study group consisted of a sample of 32 untreated subjects with normal occlusions. Lateral cephalograms were analyzed at 11 consecutive stages, from 6-13 and from 15-17 years of age. A customized cephalometric analysis was used to measure 53 variables. Statistical comparisons of gender-specific differences were performed by means of Mann-Whitney U tests.Anterior and posterior cranial base lengths, midfacial length as well as mandibular length were recorded to be significantly larger in male subjects at the age of 6 years. For most of the linear measurements, significantly larger craniofacial distances were recorded in males from the age of 15 years onward. There were no statistically significant gender differences with regard to most angular measurements at subsequent age groups. Soft tissue analysis revealed flatter profiles in females than in males from the age of 10-11 years onward, while age-dependent changes in the soft tissue profile were similar in both genders.In untreated subjects with normal occlusion craniofacial development of the hard and soft tissues can be considered age- and gender-dependent. Therefore age- and gender-specific differences of linear craniofacial distances should be taken into account for diagnosis and treatment planning in children and adolescents. The present results can be used as reference values for children and adolescents of German origin.

Malocclusion and caries prevalence: is there a connection in the primary and mixed dentitions?

The purpose of this epidemiological cross-sectional study was to determine the prevalence of malocclusion and caries in children and to investigate whether a relationship exists between prevalence of caries and studied malocclusion. The study consisted of 8,864 preschool and schoolchildren with primary dentitions (mean age 4.5 years) and mixed dentitions (mean age 8.9 years). 1997 WHO dental caries criteria were applied to both groups. The existence of an increased caries risk was deducted from the dmft and DMFT indices related to age. Malocclusion in primary and mixed dentitions was classified into seven types. Fifty-seven percent of all children had some form of malocclusion. Prevalence of malocclusion increased and was significantly greater in the mixed dentition sample (p < 0.001) than in the primary dentition sample. Seventy-four percent of children with primary dentitions and 23% of children with mixed dentitions had zero dmft and DMFT scores. Mean dmft indices in subjects with primary and mixed dentitions were 1.02 and 1.53, respectively. No positive correlation between prevalence of caries and malocclusion could be established in the sub sample with primary teeth only. However, statistically significant parallelism in prevalence of malocclusion and caries were found for posterior cross-bite (p= 0.050) and mandibular overjet (p= 0.013) in children with mixed dentitions.

Comparative immunophenotypic study of lichen sclerosus: epidermotropic CD57+ lymphocytes are numerous--implications for pathogenesis.

To characterize the immunophenotype of inflammatory cells in lichen sclerosus (LS), we performed a comparative case control study using one- and two-color immunohistochemistry and the nitro blue tetrazolium (NBT) reaction. Study material consisted of 100 biopsies from patients with LS or from 12 control groups consisting of inflammatory, scarring, and depigmenting cutaneous disorders. In addition, fresh tissue was sampled from four vulvectomy specimens for NBT testing. The typical inflammatory infiltrate of LS contained numerous epidermotropic CD3+, CD8+, CD57+ cells, increased intraepidermal HLA-DR+ cells, and a dermal infiltrate rich in CD8+, CD57+, HLA-DR+, and CD68+ inflammatory cells. Comparing LS to the 12 control groups, epidermotropic CD57+ lymphocytes independently predicted LS (P = 0.006, logistic regression, multivariate analysis). Among the 12 control groups, only specimens of the inflammatory stage of morphea exhibited numerous dermal CD57+ lymphocytes. Two-color immunohistochemistry confirmed the CD3+/CD8+CD57+ and CD3+/ CD8+/CD57+HLA-DR+ epidermotropic and dermal lymphocytic phenotypes and the dermal macrophage CD68+HLA-DR+ phenotype. In LS, the NBT reaction revealed evidence of superoxide production associated with CD68+HLA-DR+ cells. Expansion of CD8+CD57+lymphocytes is associated with viral infections, autoimmune disease, malignancies, and transplantation and is suspected to be the result of chronic excessive antigen challenge. In these pathologic states, CD8+CD57+ lymphocytes (as terminally differentiated, antigen-specific T cells) participate in the suppression of cytolytic activity to limit tissue damage. In LS, activated macrophages and lymphocytes indicate persistent antigen-driven inflammation. LS's numerous CD8+CD57+ lymphocytes may be either the mediators or the consequence of its hallmark sclerosis.

The influence of caries of the deciduous teeth upon development of the dentition in patients with cleft lip, jaw and palate.

In 417 children aged 3 to 8 years with cleft lip, jaw and palate, the prevalence of caries and the degree of treatment of deciduous teeth were compared with those of 258 cleft patients who had received preventive treatment. The prevalence of caries was reduced by more than 50% by preventive treatment. A statistically significant effect of the cleft on the prevalence of caries could not be demonstrated. Longitudinal investigations on casts showed reduction of leeway spaces in consequence of caries and early extractions. Recommendations are made for curative pediatric dental treatment.

Functional implications of genetic interactions between genes encoding small GTPases involved in vesicular transport in yeast.

Ras-related, guanine nucleotide-binding proteins of the Ypt/Rab family play a key role at defined steps in vesicular transport, both in yeast and in mammalian cells. In yeast, Ypt1p has an essential function late in endoplasmic reticulum (ER) to Golgi transport, and the redundant Ypt31/Ypt32 GTPases have been proposed to act in transport through and/or from the Golgi. Here we report that mutant alleles of YPT31 and YPT32, whose gene products have a reduced affinity for GTP, are able to suppress the dominant lethal phenotype of YPT1(N121I). Co-expression of YPT1(N121I) and the suppressor YPT31(N126I) allow essentially undisturbed secretory transport in the absence of the respective wild-type GTPases. Such mutant cells massively overaccumulate 60-100 nm vesicles and are heat sensitive. It appears likely that the mutant GTPases, which are defective in nucleotide binding, compete for the binding of common interacting protein(s). These and other genetic interactions between YPT1, YPT31/32, ARF1 and SEC4 described here strongly support the view that Ypt31p and Ypt32p have a central, Golgi-associated function in anterograde or retrograde transport.

Specific interaction of the yeast cis-Golgi syntaxin Sed5p and the coat protein complex II component Sec24p of endoplasmic reticulum-derived transport vesicles.

The generation of transport vesicles at the endoplasmic reticulum (ER) depends on cytosolic proteins, which, in the form of subcomplexes (Sec23p/Sec24p; Sec13p/Sec31p) are recruited to the ER membrane by GTP-bound Sar1p and form the coat protein complex II (COPII). Using affinity chromatography and two-hybrid analyses, we found that the essential COPII component Sec24p, but not Sec23p, binds to the cis-Golgi syntaxin Sed5p. Sec24p/Sed5p interaction in vitro was not dependent on the presence of [Sar1p.GTP]. The binding of Sec24p to Sed5p is specific; none of the other seven yeast syntaxins bound to this COPII component. Whereas the interaction site of Sec23p is within the N-terminal half of the 926-aa-long Sec24p (amino acid residues 56-549), Sed5p binds to the N- and C-terminal halves of the protein. Destruction by mutagenesis of a potential zinc finger within the N-terminal half of Sec24p led to a nonfunctional protein that was still able to bind Sec23p and Sed5p. Sec24p/Sed5p binding might be relevant for cargo selection during transport-vesicle formation and/or for vesicle targeting to the cis-Golgi.

The Saccharomyces cerevisiae SOP1 and SOP2 genes, which act in cation homeostasis, can be functionally substituted by the Drosophila lethal(2)giant larvae tumor suppressor gene.

By complementation of a salt-sensitive mutant of Saccharomyces cerevisiae, we cloned the SOP1 gene, encoding a 114.5-kDa protein of 1033 amino acids. Cells deleted for SOP1 exhibited sensitivity to sodium stress, but showed no sensitivity to general osmotic stress. Following exposure of sop1Delta cells to NaCl stress, the intracellular Na+ level and the Na+/K+ ratio rose to values significantly higher than in wild type cells. Deletion of SOP2, encoding a protein sharing 54% amino acid identity with Sop1p, produced only slight Na+ sensitivity. Cells carrying a sop1Deltasop2Delta double deletion became, however, hypersensitive to Na+ and exhibited increased sensitivity also to Li+ and K+, suggesting involvement of both SOP1 and SOP2 in cation homeostasis. The predicted amino acid sequences of Sop1p and Sop2p show significant homologies with the cytoskeletal-associated protein encoded by the Drosophila lethal(2)giant larvae tumor suppressor gene. Immunolocalization of Sop1p revealed a cytoplasmic distribution and cell fractionation studies showed that a significant fraction of Sop1p was recovered in a sedimentable fraction of the cytosolic material. Expression of a Drosophila l(2)gl cDNA in the sop1Deltasop2Delta strain partially restored the Na+ tolerance of the cells, indicating a functional relationship between the Sop proteins and the tumor suppressor protein, and a novel function in cell homeostasis for this family of proteins extending from yeast to human.

Vulvar lichen sclerosus and squamous cell carcinoma: a cohort, case control, and investigational study with historical perspective; implications for chronic inflammation and sclerosis in the development of neoplasia.

The histological changes of lichen sclerosus (LS) are frequently found in association with vulvar squamous cell carcinoma (SCC). The importance of chronic inflammation and scarring in oncogenesis is well recognized. Thirty-two patients with symptomatic vulvar LS and 60 with vulvar SCC were studied. Paraffin sections of vulvar LS, and three controls groups (acute scars, normal vulva, and vulvar lichen simplex chronicus [LSC]) were investigated with a panel of seven tissue markers and for DNA content in areas without vulvar intraepithelial neoplasia (VIN). All published cases to date of vulvar LS associated with SCC were reviewed. Of the cohort of symptomatic vulvar LS patients (mean/median age, 60 years), 9% developed VIN lesions and 21% invasive SCC; symptomatic LS preceded the carcinoma by a mean of 4 years (range, 1 to 23 years). Second and third primary tumors developed in three of these patients. Of the series of 60 patients presenting with vulvar SCCa, the clinical setting and histological features of SCCs associated with LS were significantly distinctive compared with SCCas without LS: SCCs associated with LS occurred in an older age-group (74 v 65 years; P = .01), were located on the clitoris (41% v 5%; P = .003), were of conventional SCCa type (85% v 57%; P = .02), were associated with a prominent fibromyxoid stromal response (46% v 10%; P = .004), were not associated with VIN 3 (SCC in situ) (5% v 67%; P = .02) and diffusely expressed tumor suppressor gene product p53 (43% v 19%; P = .01) and cytokine TGF-beta (33% v 9%; P = .05). The epidermis of vulvar LS was similar to that of acute scars and differed significantly compared with normal vulva with respect to keratinocytic expression of markers to keratin AE 1, involucrin and filaggrin, epidermal thickness (0.13 mm [LS] v 0.05 mm [normal]; P < .03), and proliferative index by PCNA and Mib-1 labeling (53/60 [LS] v 15/19 [normal] per 200 basal cells [bc]; P < .003). Vulvar LS showed significantly higher expression of p53 than all three control groups (80 [LS] v 3 [normal]/44 [acute scar]/28 [LSC] per 200 bc; P < .008), and aneuploidy (33% v diploid controls) in the absence of VIN. Comparing LS with and without associated SCCa found significant increases in age of patients (74 v 66 years; P = .001), and DNA aneuploidy (52% v 11%; P = .0001) and no differences in epidermal thickness, sclerotic thickness, proliferative index, or p53 expression. However, those cases of LS with an aneuploid DNA content showed significantly elevated p53 expression (88 v 60/200 bc; P = .01) and epidermal thickness (0.16 v 0.11 mm; P = .005) compared with LS with a diploid DNA content. Review of published cases supports an association between LS and vulvar SCC. The phenomenon of chronic inflammation and scarring giving rise to carcinoma has been well documented. Vulvar lichen sclerosus (LS) is an inflammatory dermatosis characterized by clinicopathologic persistence and hypocellular fibrosis (sclerosis). A subset of vulvar SCCs is significantly associated with the presence of LS and diffusely express the p53 gene product. Keratinocytes affected by LS show a proliferative phenotype and can exhibit markers of neoplastic progression such as increased p53 expression and DNA aneuploidy. As a chronic scarring inflammatory dermatosis, vulvar LS could act as both "initiator and promoter" of carcinogenesis, explaining the frequent coexistence of these diseases. Because keratinocytes of LS significantly express tumor suppressor gene p53 protein, the p53 gene may be involved early in this proposed pathway of carcinogenesis.

High-affinity binding of the yeast cis-Golgi t-SNARE, Sed5p, to wild-type and mutant Sly1p, a modulator of transport vesicle docking.

Docking of ER-derived vesicles to the cis-Golgi compartment in yeast requires vesicle and target membrane receptors (v-SNAREs and t-SNAREs) and the GTPase Ypt1p. The t-SNARE Sed5p is complexed with Sly1p in vivo. The mutant form Sly1-20p rescues Ypt1p-lacking cells from lethality, suggesting an inhibitory function of Sly1p in v-SNARE/t-SNARE interaction. Using surface plasmon resonance spectroscopy, we found that Sed5p binds Sly1p and Sly1-20p with equally high affinity (K(D) = 5.13 x 10(-9) M and 4.74 x 10(-9) M, respectively). Deletion studies show that the N-terminal half of Sly1p rather than the C-terminus (harbouring the E532K substitution in Sly1-20p) is most critical for its binding to Sed5p. These data appear to argue for an active rather than an inhibitory role of Sly1p in vesicle docking.

Theta-like activity in hippocampal formation slices: the effect of strong disinhibition of GABAA and GABAB receptors.

The involvement of GABAA and GABAB receptors in neural mechanisms responsible for the production of theta rhythms in hippocampal formation (HPC) slices is addressed in the present study. In a number of papers published in the last decade, we have demonstrated that theta-like activity can be successfully recorded in the limbic cortex maintained in vitro when the cholinergic agonists, acetylcholine, carbachol or muscarine, were added to the bath. Recently, we have also shown a strong GABAA modulation of the cholinergic-induced in vitro theta-like activity. This study presents a report of the first demonstration of in vitro theta-like field responses induced a consequence of simultaneously inhibiting hippocampal GABAA and GABAB receptors. HPC slices (350 microns) were maintained in a gas-liquid interface chamber (35 degrees C). Theta-like activity was induced in the presence of bath perfusion of bicuculline (GABAA antagonist) and 2-hydroxysaclophen (GABAB antagonist). This in vitro induced field response was antagonized both by muscimol (GABAA agonist) and baclophen (GABAB agonist). In addition, the experiments presented here revealed that bicuculline/2-hydroxysaclophen-induced in vitro theta-like activity also had a strong cholinergic M1 involvement: it was abolished by hemicholinium-3 (choline transport blocker) and pirenzepine (specific antagonist of M1 receptor), but not by gallamine (specific antagonist of M2 receptor). The results of the present study provided further evidence for a strong GABAergic/cholinergic interaction in the neural mechanism responsible for production of theta-like activity in the hippocampal formation slices.

The spontaneous theta rhythm recorded from the hypothalamus posterior in the cat in vivo.

Rhythmical slow activity (theta) was mapped in the hypothalamic region in freely moving cats. We recorded well synchronized and high amplitude theta rhythm in the medial part of the hypothalamus posterior area. The EEG recordings made from lateral part of this hypothalamic region contained only irregular activity. These findings support earlier observations concerning the topography of hippocampal formation desynchrony and synchrony system. The observations of the present study also suggest that the hypothalamus posterior area is actively involved in the mechanisms responsible for generating theta oscillations in the cat.

The rhythmic slow activity recorded from entorhinal cortex in freely moving cats.

The relationships between the entorhinal cortex (EC) and the hippocampal formation (Hipp) field potentials were examined in the present study. The detailed analyses of the signal let us group the patterns of theta appearance into three categories: (1) Theta rhythm dominating in both recordings from the EC and from the Hipp (2) Theta rhythm dominating in the Hipp with irregular activity in the EC (3) Theta rhythm dominating in the EC with irregular activity in the Hipp. These findings provide the evidence for the intrinsic generator of theta rhythm to be localized in entorhinal cortex in cats.

Muscarinic (M1) mediation of carbachol-induced theta in the cat entorhinal cortex in vitro.

Entorhinal cortex slice preparations obtained from the cat exhibited theta rhythm during perfusion with 50 microM carbachol. The effect of carbachol was antagonized by the muscarinic blocker atropine sulphate, but not by hexamethonium and mecamylamine, which are antagonists of the nicotinic receptor. Further analysis of the pharmacological profile of these carbachol-induced theta oscillations showed that M1 receptor subtype to be involved in mediation of this EEG activity: the theta rhythm was antagonized by the M1 receptor blocker pirenzepine, but was unaffected by gallamine, an antagonist of the M2 receptor subtype.

[Metrical studies of the pharynx in patients with cheilognathopalatoschisis and their significance for nasal breathing]. / Metrische Untersuchungen des Pharynx bei Patienten mit Lippen-Kiefer-Gaumen-Spalten und deren Stellenwert für die Nasenatmung.

This study compares the topography of the pharyngeal structures of 115 patients with isolated and combined cleft palates with the topography of the pharyngeal structures of 115 patients without cleft palates. The study revealed that the sagittal dimensions in the cleft palate patients are significantly reduced. The maximum closeness can be observed at the lower margin of the velum. In order to clarify growth dependent change of the sagittal pharynx dimensions the growth of the pharynx of 61 cleft palate patients was documented at 4 different points in time between age 7 and age 18. Over this period of growth only a slight increase in the sagittal plane of the pharyngeal area was observed. Especially conspicuous was the stagnation of the distance of the lower margin of the velum to the posterior pharyngeal wall. An increase in air passage resistance, which is the cause of incompetent breathing with all its consequences for growth and dentition development, is derived from these topographical relationships. The study concludes that the postoperative functional competence of the velum is a conditio sine qua non not only in relation to speech but also in relation healthy breathing.

A small GTP-binding protein from Arabidopsis thaliana functionally complements the yeast YPT6 null mutant.

A clone designated A.t.RAB6 encoding a small GTP-binding protein was isolated from a cDNA library of Arabidopsis thaliana leaf tissue. The predicted amino acid sequence was highly homologous to the mammalian and yeast counterparts, H.Rab6 and Ryh1/Ypt6, respectively. Lesser homology was found between the predicted Arabidopsis protein sequence and two small GTP-binding proteins isolated from plant species (44% homology to Zea mays Ypt1 and 43% homology to Nicotiana tabacum Rab5). Conserved stretches in the deduced amino acid sequence of A.t.Rab6 include four regions involved in GTP-binding, an effector region, and C-terminal cysteine residues required for prenylation and subsequent membrane attachment. Northern blot analysis demonstrated that A.t.Rab6 mRNA was expressed in root, leaf, stem, and flower tissues from A. thaliana with the highest levels present in roots. Escherichia coli produced histidine-tagged A.t.Rab6 protein-bound GTP, whereas a mutation in one of the guanine nucleotide-binding sites (asparagine122 to isoleucine) rendered it incapable of binding GTP. Functionally, the A.t.RAB6 gene was able to complement the temperature-sensitive phenotype of the YPT6 null mutant in yeast. The isolation of this gene will aid in the dissection of the machinery involved in soluble protein sorting at the trans-Golgi network of plants.

[Ear manifestations in adolescents after closure of lip-jaw-palate- or isolated palatal clefts]. / Ohrbefunde bei Jugendlichen nach Verschluss einer Lippen-Kiefer-Gaumen- oder isolierten Gaumenspalte.

We examined two groups of teenagers who had been surgically treated as small children for cleft palate. Most patients were between 13 and 21 years of age. One group had been looked after by the Dept. of Orthodontics at the University of Erlangen-Nürnberg, the other by the Dept. of Orthodontics at the University of Rostock. There were differences in sequence and time of the surgical closure between the two departments. Additionally, 60% of the people treated in Rostock had a velopharyngoplastic, which was rarely the case in Erlangen. In both groups only a few patients had been seen by an ENT-doctor regularly. Only some patients had been previously treated with tubes. There was one patient in each group with a bilateral, most likely genetically determined, sensorineural hearing loss. In Erlangen we examined 66 teenagers (132 ears). Six ears had been previously treated with one or more tympanoplasties. 10 ears needed further treatment due to a seromucotympanon, adhesions, perforations of the ear drum, suspicion of cholesteatoma or insufficient improvement of hearing after previous tympanoplasty. Another 18 ears showed signs of former inflammations. The control group in Rostock included 63 patients (i.e. 126 ears). 14 of the ears examined had undergone one or more tympanoplasties previously. 13 other ears needed further treatment for seromucotympanon, adhesions, perforations of the ear drum, insufficient improvement of hearing after tympanoplasty or cholesteatoma. Residuals due to prior inflammations were found in another 26 ears. Possible reasons for the different occurrence of middle ear problems in both groups are discussed.

Bacterial L-serine dehydratases: a new family of enzymes containing iron-sulfur clusters.

Two families of enzymes are described which catalyse identical chemical reactions but differ in their prosthetic groups and hence in their mechanism of action. One family, the pyridoxal-5'-phosphate (PLP)-dependent L-threonine dehydratases, also use L-serine as substrate. The other, hitherto unrecognized family is the iron-dependent, highly specific bacterial L-serine dehydratases. It has been shown that L-serine dehydratase from the anaerobic bacterium Peptostreptococcus asaccharolyticus contains an iron-sulfur cluster but no PLP. A mechanism for the dehydration of L-serine which is similar, but not identical, to that of the dehydration of citrate catalysed by aconitase is proposed.