Direct biological effects of fractional ultrapulsed CO2 laser irradiation on keratinocytes and fibroblasts in human organotypic full-thickness 3D skin models.

Molecular effects of various ablative and non-ablative laser treatments on human skin cells-especially primary effects on epidermal keratinocytes and dermal fibroblasts-are not yet fully understood. We present the first study addressing molecular effects of fractional non-sequential ultrapulsed CO2 laser treatment using a 3D skin model that allows standardized investigations of time-dependent molecular changes ex vivo. While histological examination was performed to assess morphological changes, we utilized gene expression profiling using microarray and qRT-PCR analyses to identify molecular effects of laser treatment. Irradiated models exhibited dose-dependent morphological changes resulting in an almost complete recovery of the epidermis 5 days after irradiation. On day 5 after laser injury with a laser fluence of 100 mJ/cm2, gene array analysis identified an upregulation of genes associated with tissue remodeling and wound healing (e.g., COL12A1 and FGF7), genes that are involved in the immune response (e.g., CXCL12 and CCL8) as well as members of the heat shock protein family (e.g., HSPB3). On the other hand, we detected a downregulation of matrix metalloproteinases (e.g., MMP3), differentiation markers (e.g., LOR and S100A7), and the pro-inflammatory cytokine IL1α.Overall, our findings substantiate the understanding of time-dependent molecular changes after CO2 laser treatment. The utilized 3D skin model system proved to be a reliable, accurate, and reproducible tool to explore the effects of various laser settings both on skin morphology and gene expression during wound healing.

Controversial constitutive TSHR activity: patients, physiology, and in vitro characterization.

G protein-coupled receptors constitute a large family of transmembrane receptors, which activate cellular responses by signal transmission and regulation of second messenger metabolism after ligand binding. For several of these receptors it is known that they also signal ligand-independently. The G protein-coupled thyroid stimulating hormone receptor (TSHR) is characterized by a high level of constitutive activity in the wild type state. However, little is known yet concerning the physiological relevance of the constitutive wild type TSHR activity. Certainly, knowledge of the physiological relevance of constitutive wild type receptor activity is necessary to better understand thyroid physiology and it is a prerequisite for the development of better therapies for nonautoimmune hyperthyroidism and thyroid cancer. Based on a literature search regarding all published TSHR mutations, this review covers several mutations which are clearly associated with a hyperthyroidism-phenotype, but interestingly show a lack of constitutive activity determined by in vitro characterization. Possible reasons for the observed discrepancies between clinical phenotypes and in vitro characterization results for constitutive TSHR activity are reviewed. All current in vitro characterization methods for constitutive TSHR mutations are "preliminary attempts" and may well be revised by more comprehensive and even better approaches. However, a standardized approach for the determination of constitutive activity can help to identify TSHR mutations for which the investigation of additional signaling mechanisms would be most interesting to find explanations for the current clinical phenotype/in vitro discrepancies and thereby also define suitable methods to explore the physiological relevance of constitutive wild type TSHR activity.

[War trauma and PTSD among German war survivors. A comparison of former soldiers and women of World War II]. / Kriegstraumata und PTBS bei deutschen Kriegsüberlebenden. Ein Vergleich ehemaliger Soldaten und Frauen des Zweiten Weltkriegs.

Stressful war experiences can cause posttraumatic stress disorder (PTSD) in survivors. To what extent were the soldiers and young women of World War II affected by PTSD symptoms over the course of their lives? Do these men and women differ in the traumatic experiences and PTSD symptom severity? To investigate these questions 52 male and 20 female Germans aged 81-95 years were recruited through newspaper advertisements and notices and interviewed regarding war experiences and PTSD symptoms. Of the men 2% and 7% met the criteria for current and lifetime PTSD diagnoses, respectively, as compared to 10% and 30% of the women, respectively. Using multiple linear regression a dose-response relationship between the number of trauma types experienced and PTSD symptom severity could be demonstrated. The slope of the regression curve was steeper for women than for men. When controlling for the number of different traumatic experiences women reported a significantly higher severity of PTSD symptoms than men. It is presumed that this difference in severity of symptoms can be attributed to qualitative differences in the type of traumatic stress factors during the war. The present study provides evidence that even today people continue to be affected by PTSD symptoms due to events which occurred during World War II; therefore, during patient contact with this age group the war experiences specific to each individual need to be considered as potential moderators of symptoms.

Late manifestation of subclinical hyperthyroidism after goitrogenesis in an index patient with a N670S TSH receptor germline mutation masquerading as TSH receptor antibody negative Graves' disease.

In 27 families with familial non-autoimmune hyperthyroidism (FNAH) reported up to date, the onset of hyperthyroidism varies from 18 months to 60 years. Also the manifestation of goitres is variable in these families. A 74-year-old woman first presented at the age of 69 years with tachyarrhythmia and hypertension. After initial treatment of her hypertension and oral anticoagulation for her intermittent atrial fibrillation, a thyroid workup revealed a suppressed TSH and normal fT3 and fT4. TPO, TSH receptor (TSHR), and thyroglobulin antibodies were negative. Thyroid ultrasound revealed a thyroid volume of 102 ml with several nodules with diameters of up to 2.6 cm right and up to 1.8 cm left. Scintigraphy showed a homogeneous Technetium-99 m ((99 m)Tc) uptake of 1.27%. She was subsequently treated with 1 GBq radioiodine ((131)I). At the age of 74, her thyroid function was normal and her thyroid volume decreased to 90 ml. Because of the diffuse (99 m)Tc uptake and the negative TPO, TSHR, and thyroglobulin antibodies, genetic analysis of her TSHR gene was performed, in spite of her negative family history for hyperthyroidism. Sequencing revealed a N670S TSHR germline mutation. Previous in vitro characterisation of this TSHR mutation suggests a weak constitutive activity, yet the experimental data are ambiguous. This case illustrates the necessity to analyse patients with hyperthyroidism accompanied by diffuse (99 m)Tc uptake and negative TPO, TSHR, and thyroglobulin antibodies for TSHR germline mutations. Moreover, it demonstrates that TSHR germline mutations may first lead to longstanding nodular goitrogenesis before the late manifestation of subclinical hyperthyroidism.

[A young fireman candidate with an abnormal chest x-ray]. / Auffälliger Röntgenthorax bei jungem Feuerwehranwärter.

An 18-year-old, asymptomatic male underwent a routine chest x-ray examination which showed a pathological result. It revealed a characteristic crescent-like shadow in the right lower lung field, resembling a muslim sword (scimitar), responsible for the name of the underlying disease. Scimitar syndrome is a rare congenital malformation with a wide clinical spectrum ranging from asymptomatic to severe disturbances manifesting in childhood. The typical findings are hypoplasia of the right lung with marked mediastinal shift to the right and dextrocardia, reduced right pulmonary artery perfusion with maintained bronchial connection, anomalous arterial supply to the right lower lobe from the aorta and abnormal draining of the enlarged right pulmonary vein into the pulmonary circulation, producing the characteristic radiographic "scimitar sign". The development of right ventricular failure due to long-standing right ventricular overload and recurrent respiratory infections depend on the severity of anatomical abnormalities and functional disorders, sometimes requiring surgical intervention. Nowadays, sophisticated imaging techniques allow precise and rapid diagnosis of these complex abnormalities and their functional implications. The findings for our patient differed from the classic constellation, with a regular emptying of the enlarged right pulmonary vein into the left atrium, leading to a decreased burden on the central haemodynamics. In our very rare case of a scimitar syndrome variant, the functional disorders were negligible.

[Color reaction of chlorhexidine and proguanil with hypobromite]. / Zur Farbreaktion von Chlorhexidin und Proguanil mit Hypobromit.

Colour reaction of chlorhexidine and proguanil with hypobromite The antimalarial agent proguanil reacts with hypobromite to yield the red coloured (E)-3-[(4-chlorophenyl)imino]-N-isopropyl-3H-1,2,4-triazol-5-amine (6B). The structure of 6B was proven by X-ray. The red colour obtained by the test for the disinfectant chlorhexidinedihydrochloride Ph. Eur. with hypobromite is probably attributable to a corresponding chromophore.

Metal insertion into NiFe-hydrogenases.

The synthesis and the insertion of the metallocentre of NiFe-hydrogenases is a complex process, in which seven maturation enzymes plus ATP, GTP and carbamoyl phosphate are involved. The review summarizes what is known about the properties and activities of these auxiliary proteins, and postulates a pathway along which maturation may take place.

Relative allergenic potential of four proteases used as contact lens cleaners.

The general use of enzymatic cleaners for soft contact lenses has led to reports of incidents of allergic reactions in previously sensitized patients. In light of this, experiments were performed to assess the relative allergenic potential of four such enzymes: papain, subtilisin A, subtilisin B, and pancreatin. Mice immunized intraperitoneally (i.p.) with either subtilisin A or papain exhibited little serum immunoglobulin E (IgE), even after tertiary doses of protein. Primates immunized intradermally (i.d.) with the four enzymes showed various levels of response to challenges: none to papain, mild to moderate to subtilisin A and pancreatin, and severe to subtilisin B. Finally, guinea pigs instilled ocularly with enzymes weekly for 15 weeks exhibited significant hyperemia and chemosis only to the positive control protein, ovalbumin, and not to any of the enzymes tested. We conclude that three of the four enzymes tested have low potential to sensitize human beings, and, further, that animal models may find increased use in screening products of biotechnology.

Effect of in vivo and in vitro surface protein deposits on the oxygen permeability of polyhydroxyethylmethacrylate gel contact lenses.

The effect of surface protein deposits on the oxygen permeability of polyhydroxyethylmethacrylate ( polyHEMA ) gel contact lenses was studied using: (1) previously worn lenses with protein deposits accumulated in vivo, and (2) new lenses with protein deposited in vitro. The oxygen permeability (Dk) of the protein-deposited in vivo lenses was determined before and after cleaning, and that of the new lenses before and after in vitro protein deposition. For both groups of lenses, the oxygen permeability did not change significantly after either cleaning (in vivo lenses) or protein deposition (in vitro lenses). It is concluded that surface protein deposits on polyHEMA lenses do not alter the oxygen permeability of the lenses.

Calcium and protein in tears: diurnal variation.

With the use of an automatic titrator for calcium and a microcolorimetric assay for protein, diurnal variations in tear calcium and total protein values were studied in nine subjects during a ten-day period. Tear calcium and total protein concentrations were found to be individualized functions that were dependent on the sampling time and sampling day. Prolonged eye closure resulted in an approximate twofold increase in both tear calcium and total protein concentrations. The concentrations of both species remained relatively constant during the day between 8 AM and 5 PM, exhibiting few substantive variations. Diurnal changes in tear calcium and protein values closely paralleled one another. It was found that the tear concentrations of these two species could be related with a simple linear function, which suggests that calcium binding and transport by tear protein may be occurring.

Calcium in tears and contact lens wear.

Tear calcium levels were measured in 27 normal subjects. An instrumental technique that required only 5- to 10-microL tear samples was used for this purpose. A mean of 2.11 +/- 49 (SD) mg/dl of calcium with a range of 1.05 to 3.49 mg/dl was determined for 67 tear samples from 27 subjects. This is in excellent agreement with more elaborate and previous methods. Various factors involved in tear calcium levels were also studied. Tear calcium levels were not significantly affected by the normal walking-hour wear of either hard or soft contact lenses among 22 women aged 19 to 35 years. Tear calcium day-to-day and diurnal variations were found for five of five subjects in a three-day study. Further studies of these variable phenomena are indicated.