Fateful triad of reactive oxygen species, mitochondrial dysfunction and lipid accumulation is associated with expression outline of the AMP-activated protein kinase pathway in bovine blastocysts.

Low cryotolerance is considered as the major drawback of in vitro-produced bovine embryos and is frequently associated with a triad encompassing increased cytoplasmic lipid accumulation, enhanced levels of reactive oxygen species (ROS) and mitochondrial dysfunction. The aim of the present study was to explore the role of the AMP-activated protein kinase (AMPK) pathway in the process resulting such phenotypes. Comparative analysis under different environmental conditions revealed downregulation of AMP-activated protein kinase cytalytic subunit 1alpha (AMPKA1), peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC1A) and carnitine palmitoyltransferase 1 (CPT1) genes and upregulation of acetyl-CoA carboxylase α (ACC). In contrast, the presence of fatty acids within the culture medium resulted in a distinct molecular profile in the embryo associated with enhanced levels of ROS, mitochondrial dysfunction and elevated lipid accumulation in bovine embryos. Because AMPKA1 regulates PGC1A, CPT1 and ACC, the results of the present study reveal that AMPK in active its form is the key enzyme promoting lipolysis. Because AMPK1 activity is, in turn, controlled by the AMP : ATP ratio, it is possible to speculate that excessive uptake of exogenous free fatty acids could increase cellular ATP levels as a result of the disturbed ß-oxidation of these external fatty acids and could therefore bypass that molecular feedback mechanism. Subsequently, this condition would cause enhanced generation of ROS, which negatively affect mitochondrial activity. Both enhanced generation of ROS and low mitochondrial activity are suggested to enhance the accumulation of lipids in bovine embryos.

Molecular signatures of bovine embryo developmental competence.

Assessment of the developmental capacity of early bovine embryos is still an obstacle. Therefore, the present paper reviews all current knowledge with respect to morphological criteria and environmental factors that affect embryo quality. The molecular signature of an oocyte or embryo is considered to reflect its quality and to predict its subsequent developmental capacity. Therefore, the primary aim of the present review is to provide an overview of reported correlations between molecular signatures and developmental competence. A secondary aim of this paper is to present some new strategies to enable concomitant evaluation of the molecular signatures of specific embryos and individual developmental capacity.

Expression of the Fc-receptor for IgE (Fc epsilon RII, CD23) on alveolar macrophages in extrinsic allergic alveolitis.

Expression of the Fc receptor for IgE (Fc epsilon R) was analyzed on alveolar macrophages (AM) in 10 patients with extrinsic allergic alveolitis (EAA) compared with 10 patients with sarcoidosis and to 6 apparently healthy controls. By using the anti-Fc epsilon RII mAb M-L25 in immunocytochemistry experiments, we found that greater than 60% of AM in 10 of 10 patients with EAA were strongly positive, as evidenced by visual analysis in light microscopy and by cytometry. By contrast, no significant staining was detected in sarcoidosis or in controls with either method. Similar results were obtained when Fc epsilon R were identified with preformed immune complexes consisting of NIP-specific human/mouse chimeric IgE antibody plus NIP-ovalbumin. Furthermore, greater than 60% of AM in patients with EAA stained positive for IgE, demonstrating that endogenous IgE is bound to the AM. Our data suggest that IgE antibodies bound to Fc epsilon RII on AM may be involved in pathophysiology of extrinsic allergic alveolitis by activation of the AM after binding of allergen to the cell surface IgE. Furthermore, with the clearcut pattern of Fc epsilon RII expression in extrinsic allergic alveolitis it may be possible to use CD23 antibodies for differential diagnosis of inflammatory lung disease.

The metabolism and pharmacokinetics of isotretinoin in patients with acne and rosacea are not influenced by ethanol.

BACKGROUND: Isotretinoin is effective in the treatment of severe acne and rosacea. Both parent drug and its main metabolite 4-oxo-isotretinoin are potentially teratogenic compounds and contain a carboxylic acid moiety. In the presence of ethanol, naturally occurring as well as synthetic retinoids also containing a carboxylic acid moiety are capable of undergoing an ethyl esterification with the metabolic formation of more lipophilic compounds with a much longer terminal half-life. OBJECTIVES: To determine if isotretinoin (13-cis-RA), its main metabolite 4-oxo-isotretinoin (4-oxo-13-cis-RA), and other possible metabolites in the presence or absence of ethanol are converted to their corresponding ethyl derivatives in patients with severe acne or rosacea after multiple isotretinoin dosing. In addition, pharmacokinetic parameters of the parent drug and its 4-oxo metabolite were determined. PATIENTS/METHODS: Eleven patients with severe acne or rosacea were treated with isotretinoin daily for 3 months and investigated pharmacokinetically during 24 h after 1 month of treatment and for up to 28 days after discontinuation of therapy. A possible influence of ethanol was evaluated using a simple self-administered questionnaire and by measuring serum ethanol levels during treatment. The concentrations of isotretinoin, 4-oxo-isotretinoin and possible ethylated and nonethylated metabolites were measured by reverse-phase high-performance liquid chromatography. RESULTS: Although seven of 11 patients had a considerable weekly alcohol intake, no endogenous synthesis of ethyl derivatives of isotretinoin, the main 4-oxo metabolite or the all-trans compounds was chromatographically detectable in any of the patients' plasma samples during the treatment period. Multiple dose pharmacokinetic data for the parent drug and its main metabolite were comparable to previous studies. CONCLUSIONS: The metabolism and pharmacokinetics of isotretinoin and its main metabolites are not influenced by ethanol during long-term isotretinoin treatment. After ceasing long-term isotretinoin therapy the recommended period of 1 month for using anticonceptive measures in fertile women seems adequate.

Nickel-63 in marine and terrestrial biota, soil, and sediment.

A previously unreported radionuclide, nickel-63 (half-life, 92 years), produced in the testing of nuclear devices, was measured in biological and environmental samples from areas of the Pacific Ocean and the eastern seaboard of the United States. The concentrations of nickel-63 are low (maximum of 163 disintegrations per minute per gram of dry weight), but this radionuclide may be a useful tracer of oceanic processes because of its long half-life and long residence time in the ocean.

Reversible MRI changes in a patient with uremic encephalopathy.

A 19-year-old patient on chronic ambulatory peritoneal dialysis experienced severe neurologic disturbances caused by uremia. Increased signal intensity was seen bilaterally in the cortical and subcortical areas of the occipital and parietal lobe on cranial magnetic resonance imaging (MRI). Insufficient peritoneal dialysis efficacy was documented and the patient was switched from peritoneal to hemodialysis. Cranial MRI indicated a marked regression of the lesions to nearly normal, confirming the diagnosis of uremic encephalopathy.

Binocular vision recovery in bilateral keratoplasty.

PURPOSE: To determine the benefit of bilateral penetrating keratoplasty with regard to binocular vision. METHODS: We compared patients who underwent corneal transplantation bilaterally with patients who had successful corneal transplantation in one eye and corneal disease in the other eye. Specifically, changes in fusion, stereopsis, and binocular vision function were analyzed. A questionnaire regarding performance of daily tasks was given. RESULTS: Patients with bilateral keratoplasty performed better in all the analyzed functions. Fusion was achieved by 81.25% in the unilateral group versus 100% in the bilateral group (p = 0.15). Stereopsis was present more in the bilateral group (100% vs. 62.5%, p = 0.008) and the quantity of stereopsis was significantly better in the bilateral group (121 seconds of arc vs. 1,284 seconds of arc, p = 0.014). 88.8% of the patients subjectively improved in daily activities after second eye surgery. CONCLUSION: There are objective and subjective improvements after bilateral penetrating keratoplasty.

Inflammatory demyelinating neuropathy in patients with end-stage renal disease receiving continuous ambulatory peritoneal dialysis (CAPD)

OBJECTIVE: To report on 3 patients with inflammatory demyelinating peripheral neuropathy in strong temporal coincidence with the initiation of peritoneal dialysis (PD) therapy. SETTING: Nephrology and Neurology Department of the University Hospital, Munich, Germany. PATIENTS: Three patients with end-stage renal failure presented with the clinical picture of inflammatory demyelinating peripheral neuropathy within 4 to 10 weeks after start of continuous ambulatory peritoneal dialysis (CAPD). They had acute or subacute onset of lower extremity or generalized weakness, diminished reflexes, elevated spinal fluid protein levels, and signs of demyelinating neuropathy on electrophysiological testing. MEASURES: Clinical follow-up, nerve conduction studies, cerebral spinal fluid (CSF). RESULTS: All patients did not improve under intensified PD therapy but took profit from immunomodulatory therapy. One bed-bound patient improved after change to hemodialysis and showed complete remission after renal transplantation. CONCLUSION: Because of strong temporal coincidence, a causal relationship between CAPD and inflammatory demyelinating peripheral neuropathies can be suspected in these 3 patients.

Bioincompatible membranes place patients with acute renal failure at increased risk of infection.

The mortality of critically ill patients with acute renal failure (ARF) requiring hemodialysis remains high (> 50%), even though major improvements in the management of this disorder have been made. Sepsis has been identified as a major cause of death in ARF. Experimental data have suggested that bioincompatible membranes aggravate uremia induced neutrophil dysfunction. In the following two combined prospective, randomized, clinical studies, the effects of the biocompatibility of dialysis membranes on the susceptibility to infection were compared in 72 surgical and medical patients with ARF. The group of patients treated with biocompatible polyacrylonitrile membranes had a significantly lower incidence of bacterial infections and a lower relative and absolute mortality rate due to sepsis, compared to patients treated with bioincompatible cuprophane membranes. These investigations identified bioincompatibility as a factor that adversely affects the prognosis of critically ill patients with ARF.

Assessment of acute parathyroid responsiveness to high calcium dialysate in uremic patients.

The response of the parathyroid glands to dialysis-induced changes in calcium has been studied in 83 normocalcemic patients receiving maintenance hemodialysis. Predialysis concentrations of intact parathyroid hormone (iPTH) were normal in 13 patients; 70 patients had increased iPTH levels of different degrees. All patients had analytically significant changes in total serum calcium (2.3 +/- 0.2 vs 2.8 +/- 0.2 mmol/l) without additional parenteral calcium loading during a standard dialysis session with a dialysis solution of 1.75 mmol calcium chloride. During hemodialysis the concentration of iPTH fell in 77 patients and was normalized in 48 of these patients. The remaining 6 patients showed no or an inadequate response (less than 30% suppression of predialysis iPTH) to the increase in Ca. Parathyroidectomy was performed in patients with refractory iPTH secretion. Histological evidence of marked parathyroid hyperplasia was obtained in all these patients. All patients with normal or partial responsiveness of the parathyroids tolerated at least small doses of 1.25 (OH)2-vitamin D3. Quantification of predialysis and postdialysis iPTH concentrations is a useful and simple method for identifying those uremic patients with true autonomy of the parathyroid glands at an early stage.

The ecosystem study on Rongelap Atoll.

During the 1950's and 1960's, the Laboratory of Radiation Biology at the University of Washington carried out an intensive study of this Atoll, which was contaminated with radioactive fallout from the "Bravo shot" in 1954. This study involved many aspects of the environment and the plant and animal life: soils, land plants, marine life, birds, geology and hydrology, and human diets as well. In much of the research, the fortuitiously present radioactive isotopes, especially 137Cs and 90Sr, were tracers. Although the term "ecosystem study" was not in vogue at that time, it is clear that this was an early use of the ecosystem approach. Soil types and their development, the distribution of mineral elements in plants and soils, including predominant radionuclides, distribution and growth of native terrestrial plants in relation to topography and salinity, some aspects of the human diets, micronutrient nutrition of the coconut palm, island and islet development and stability, were given attention in the studies. Some of the findings in the various areas of study will be presented and discussed.

[What is established in the therapy of kidney diseases?]. / Was ist gesichert in der Therapie der Nierenkrankheiten?

The established therapy for glomerular disorders of the kidney is outlined in a short review. Before initiating therapy, the physician must decide if the underlying disease is of primary or secondary type. The nephrotic syndrome and progressive renal insufficiency can result in both conditions. The necessary dietary and pharmacological principles in the therapy of these syndromes are discussed. Finally, the clinical and therapeutic aspects of secondary glomerulopathy are described with selected examples.

[Porphyria cutanea tarda and hepatitis C virus infection]. / Porphyria cutanea tarda og Hepatitis C virus-infektion.

Porphyria cutanea tarda (PCT) is a rare metabolic disorder characterized by an abnormal porphyrin metabolism and typical cutaneous lesions. Recently a strong association between PCT and hepatitis C virus (HCV) has been proposed. Studies in the south of Europe have shown high prevalences (53 to 91%) of HCV markers in patients with sporadic PCT. Six characteristic case stories are presented. We suggest routine hepatitis C serological testing in all patients with PCT, as HCV infection is a major predisposing factor for the disease.

[Perioral dermatitis in children under steroid inhalation therapy]. / Perioral dermatitis hos børn i steroidinhalationsbehandling.

Topically applied corticosteroid is a known provoking factor in perioral dermatitis. In recent years inhaled corticosteroids have become first line therapy for asthma in children. We present two characteristic cases of perioral dermatitis that developed during asthma treatment with budesonide using either a nebulizer or a spacer connected with a mask. In severe cases treatment with oral erythromycin or topical metronidazole gel may be considered. The eruption should not be confused with atopic eczema.

[Investigation of malignant melanoma in an "open house" setting]. / Undersøgelse for malignt melanom ved et "åbent hus"-arrangement.

Individuals with a self-evaluated risk of malignant melanoma were invited to an "open house" where trained dermatologists examined suspected lesions. The attendants filled in a short questionnaire relating to skin cancer risk factors. Persons with skin changes suspected for malignancy were referred for excision in the dermatological department. The clinical evaluation included dermatoscopy. Nine hundred and eleven individuals attended. Among 16 individuals referred for excision one malignant melanoma, one lentigo maligna melanoma and two in situ melanomas were histologically verified. Furthermore, 21 basal cell carcinomas were identified clinically. The most frequent clinical diagnoses were melanocytic naevi and seborrhoic keratoses.

G6PDH-activity in equine oocytes correlates with morphology, expression of candidate genes for viability, and preimplantative in vitro development.

Efficiencies for in vitro production of equine embryos are still low due to highly variable developmental competences of equine immature oocytes. In contrast to the equine, in vitro developmental competence of immature oocytes has been predicted successfully by the activity of glucose-6-phosphate dehydrogenase (G6PDH) indicated by brilliant cresyl blue (BCB) dye in a range of different species. Therefore, the aim of the present study was to test the association between G6PDH activity in equine oocytes with: (1) cumulus morphology and oocyte properties in terms of diameter and volume; (2) maturational competence; (3) gene expression of certain molecular markers; and (4) in vitro embryo development after intracytoplasmic sperm injection. Equine oocytes were exposed to BCB stain and were classified as BCB+ or BCB- according to their ability to convert the dye from blue to colorless. Additionally, BCB+ and BCB- oocytes were subclassified as having a compact (Cp) or expanded (Ex) cumulus complex. As a result, BCB+ oocytes had a greater proportion of expanded cumulus oocyte complexes compared with BCB- oocytes (71.2% vs. 49.5%). Moreover, we observed a significant difference in oocyte diameter and volume between BCB+ and BCB- oocytes irrespective of cumulus morphology. BCB+ oocytes reached a higher maturation rate compared with BCB- oocytes (59.0% vs. 28.7%). Regarding the analyzed candidate genes, relative transcript abundance was significantly different for nine genes. The expression of eight genes was significantly higher (P < 0.05) for BCB+ oocytes, including ATPV6E, IF-3, TFAM, DNMT1, STAT3, Aurora-A, ODC1, and CKS2 whereas BCB- oocytes showed higher in expression of COX1. These results are in line with the observed developmental competence. Cleavage rate (45.9% vs. 29.0%) and percentage of embryos that reached the blastocyst stage (9.2% vs. 1.4%) were significantly higher for embryos derived from BCB+ oocytes compared with BCB- oocytes. In conclusion, the present study provides evidence that G6PDH-activity in immature equine oocytes is a useful predictor for subsequent in vitro developmental competence.