Palladium causes bizarre skin hyperpigmentation in long-term dihydrocodeinone 'Braun' abusers.

BACKGROUND: 'Braun' is an illegal injectable dihydrocodeinone-enriched drug mixture of semi-synthetic opioids. It is prepared by palladium-catalysed hydrogenation from codeine-containing tablets. OBJECTIVE: We aimed to characterize the dermatologic consequences of long-term abuse of 'Braun'. METHODS: Skin biopsies of two long-term 'Braun' abusers were evaluated histopathologically, immunohistochemically and ultrastructurally. Palladium skin content was assessed by X-ray fluorescence (XRF) spectrometry. RESULTS: Both patients showed generalized diffuse dark blue-grey hyperpigmentation of the skin. In both, an abnormal population of cells containing intracytoplasmic brownish granular material was identified in the papillary dermis by light microscopy. Electron microscopy revealed a dense and minimally structured material that predominantly accumulated in macrophages, fibroblasts and vascular endothelial cells. XRF analysis confirmed elevated levels of palladium in the patient's skin in comparison to healthy controls. CONCLUSION: Long-term abuse of palladium-contaminated dihydrocodeinone ('Braun') results in excessive accumulation of granular material in various dermal cell types and causes generalized diffuse skin hyperpigmentation.

Quantitation of plasmatic lysosphingomyelin and lysosphingomyelin-509 for differential screening of Niemann-Pick A/B and C diseases.

Acid sphingomyelinase deficiency (ASMd, Niemann-Pick disease A/B) and Niemann-Pick type C disease (NPC) share core clinical symptoms. Initial diagnostic discrimination of these two rare lysosomal storage diseases is thus difficult. As sphingomyelin accumulates in ASMd as well as NPC, lysosphingomyelin (sphingosylphosphorylcholine) and its m/z 509 analog were suggested as biomarkers for both diseases. Herein we present results of simultaneous LC-ESI-MS/MS measurements of lysosphingomyelin and lysosphingomyelin 509 in plasma and dried blood spots (DBS) collected from ASMd and NPC patients and suggest that the plasma but not DBS levels of the two analytes allow differential biochemical screening of ASMd and NPC.

Morphological assessment of thin basement membrane disease.

Thin basement membrane disease is more common than IgA nephropathy or Alport syndrome, which are also associated with the presence of erythrocyturia. Very few reports on the disorder are available in the Polish literature. The objective of this work was to analyze the results from 83 patients with thin basement membrane syndrome as well as to formulate a proposal of strict morphological assessment criteria for the disorder. Attention was drawn to the requirement of thickness of the lamina densa rather than the entire basement membrane thickness and a sufficiently high number of loops featuring thinned lamina densa, namely at least 80% of loops, being taken into account. Occurrence of other morphological changes associated with the disorder and clinical symptoms other than erythrocyturia was also highlighted.

Damage-Associated Molecular Patterns in the Course of Lung Cancer--A Review.

More than 20 years ago, the 'danger theory' was proposed which explains why potent immune responses with no microbial components are elicited against tissue transplants, injuries, tumours and autoimmune diseases. It states that the immune system can distinguish between dangerous and innocuous endogenous signals. In response to trauma or other types of tissue and cell damage, certain molecules that function inside the cell are released or secreted from damaged or dying cells. Such mechanisms initiate an immune response in the absence of infection. These immunostimulatory molecules were named damage-associated molecular patterns (DAMPs). In this article, we will review the available data on the influence of select DAMPs on lung cancer cells and tumour microenvironments. We will also summarize the current information regarding the interactions between lung cancer-associated DAMPs and their toll-like receptors.

Cytotoxic reaction mediators: granzymes A and B in women with ovarian cancer.

The purpose of this work was the assessment of cytotoxic reaction mediators - granzymes A and B in the serum of women with ovarian tumors. The study included 120 women with proven ovarian tumors. The control group consisted of 60 healthy women in whom no pathological changes within the reproductive system were detected. Concentrations of granzymes A and B were measured by enzyme-linked immunosorbent (ELISA) assay. The highest concentrations of the studied parameters were observed in serum of women with ovarian cancer. Moreover, the concentrations of granzymes A and B in patients with ovarian cancer were substantially increased in comparison to concentrations in patients with ovarian cystadenomas (P < 0.0001) or ovarian teratomas (P < 0.0001).

The role of Toll-like receptors and vitamin D in diabetes mellitus type 1--a review.

It is widely accepted that type 1 diabetes mellitus (T1DM) is an autoimmune disease resulting from an interaction between immunologic, genetic and environmental factors. However, the exact mechanism leading to the development of T1DM remains incomplete. There is a large body of evidence pointing towards the important role of toll-like receptor (TLR) activation and vitamin D deficiency in T1DM pathogenesis. In this article, we review the available data on the influence of TLRs' level of activation and vitamin D status on the risk of the development of T1DM in humans and rodent models. We also summarize the current information regarding the interactions between TLRs' level of activation, vitamin D status and various environmental factors, such as enteroviral infections, the gut microbiota and breastfeeding substitution, among others. Our results stipulate that vitamin D seems to protect against T1DM by reducing the TLRs' level of activation.

Clinical and molecular characterization of a family with a dominant renin gene mutation and response to treatment with fludrocortisone.

BACKGROUND: A family was identified with autosomal dominant inheritance of anemia, polyuria, hyperuricemia, and chronic kidney disease. Mutational analysis revealed a novel heterozygous mutation c.58T > C resulting in the amino acid substitution of cysteine for arginine in the preprorenin signal sequence (p.cys20Arg) occurring in all affected members. METHODS: Effects of the identified mutation were characterized using in vitro and in vivo studies. Affected individuals were clinically characterized before and after administration of fludrocortisone. RESULTS: The mutation affects endoplasmic reticulum co-translational translocation and posttranslational processing, resulting in massive accumulation of non-glycosylated preprorenin in the cytoplasm. This affects expression of intra-renal RAS components and leads to ultrastructural damage of the kidney. Affected individuals suffered from anemia, hyperuricemia, decreased urinary concentrating ability, and progressive chronic kidney disease. Treatment with fludrocortisone in an affected 10-year-old child resulted in an increase in blood pressure and estimated glomerular filtration rate. CONCLUSIONS: A novel REN gene mutation resulted in an alteration in the amino acid sequence of the renin signal sequence and caused childhood anemia, polyuria, and kidney disease. Treatment with fludrocortisone improved renal function in an affected child. Nephrologists should consider REN mutational analysis in families with autosomal dominant inheritance of chronic kidney disease, especially if they suffer from anemia, hyperuricemia, and polyuria in childhood.

Clinical evaluation of proinflammatory cytokine inhibitors (sTNFR I, sTNFR II, IL-1 ra), anti-inflammatory cytokines (IL-10, IL-13) and activation of neutrophils after burn-induced inflammation.

The study was aimed at evaluating the involvement of sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 and reactive oxygen species (ROS) in systemic inflammatory response syndrome (SIRS) development in severely burned children and at assessing the prognostic value of the immunological markers studied. The study comprised 37 patients (17 burned children and 20 controls). Serum levels of the markers determined by means of ELISA and respiratory burst of neutrophils as well as p55 and p75 tumour necrosis factor-alpha (TNF-alpha) receptor expression using flow cytometry were evaluated twice. The burned children presented significantly higher levels of IL-10 and cytokine inhibitors within the first 6-24 h after injury compared with controls (P < 0.05). The decreased oxygen metabolism of neutrophils and increased TNF-alpha receptor expression were found on admission. Moreover, a significant decrease in initially high sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 concentrations (P < 0.05) and reduced expression of TNF-alpha receptors (P < 0.05) were observed after burn therapy, whereas ROS generation evidently augmented (P < 0.05). Four of our children who developed hypovolaemic shock revealed a significantly lower ROS generation and higher concentrations of soluble TNF-alpha receptors and IL-1 ra together with IL-10, IL-13 compared with children with good outcome (P < 0.05). Our results revealed the involvement of both ROS, soluble TNF-alpha receptors and IL-1 ra in the development of SIRS in burned children; their monitoring allows for an assessment of the systemic inflammatory reaction activity. The neutrophil BURSTTEST and IL-1 ra might have been clinically helpful markers of SIRS prognosis.

A case of excessive autophagocytosis with multiorgan involvement and low clinical penetrance.

An autopsy and microscopic analyses of a 74-year-old female with a clinical history of cardiac hypertrophy and hypertension disclosed a pronounced distension of lysosomal compartment with signs of excessive autophagocytosis, predominantly in cardiomyocytes, hepatocytes and smooth muscle cells of the small intestine. The histological storage pattern did not correspond to the usual changes seen in defined lysosomal storage disorders. The amount of age-related lipopigment was low in all tissues. Confocal microscopy of liver tissue samples documented a progressive loss of mitochondrial epitopes in the distended lysosomal compartment along the porto-central axis of hepatic lobules. The possibility to detect subunit c of mitochondrial ATP synthase (SCMAS) indicated extensive intra-lysosomal degradation of mitochondria, both in hepatocytes and smooth muscle cells. The SCMAS epitope can thus be considered a valuable immunohistochemical marker of autophagocytic mitochondrial degradation. The distended lysosomes also displayed tissue specific ubiquitination. Absence of immuno-detectable p62 protein excluded aggresome formation. An inherent dysfunction of the late endosomal/lysosomal LAMP2 protein (Danon disease), was excluded on the basis of LAMP2 gene sequence analysis and LAMP2 protein levels. Whether the observed process reflects a primary dysregulation of the constitution of the autophagosomal membrane or was induced by defects in other cellular components, remains unanswered. Whatever mechanism involved, the findings should be considered relevant in differential diagnostics, despite their low clinical penetrance, should be registered and thus rendered available for future definition.

Regulatory T cells in malignant pleural effusions subsequent to lung carcinoma and their impact on the course of the disease.

Tumors exert suppressive effects on the host immune system and tumor progression can be linked to functional impairments of immune cells. Regulatory T cells (Treg) are a subpopulation of T lymphocytes and play a key role in suppressing immune responses against autoimmune diseases and cancer. The aim of the study was to investigate the prevalence of Treg in malignant and benign pleural effusions and to evaluate the relationship between Treg frequency and disease advance. Pleural effusions from 76 patients were subjected to a routine laboratory diagnosis and analyzed by conventional cytology. Biological materials were divided into three groups: malignant pleural effusions with malignant cells, effusions from patients with malignancy but without malignant cells, and non-malignant pleural effusions. The frequency of Treg in malignant pleural effusions was significantly higher compared to non-malignant effusions. In general, the increase in Treg frequency was correlated with a decrease in the percentage of lymphocytes and an increase in T CD4+ and T CD4+ CD25+ cells. The highest percentage of Treg was observed among patients with the most advanced clinical stage of lung cancer in terms of size and location of a primary tumor, T4. A Kaplan-Meier survival analysis showed a statistically significant trend towards an adverse outcome for patients representing higher Treg counts. Overall, our results support the extraordinary potential of Treg control in future anticancer therapy.

Subclinical course of adult visceral Niemann-Pick type C1 disease. A rare or underdiagnosed disorder?

We present the third case of Niemann-Pick disease type C without neurological symptoms. The patient was a 53-year-old woman without significant prior health problems who died of acute pulmonary embolism. Autopsy findings of hepatosplenomegaly, lymphadenopathy and ceroid-rich foam cells raised the suspicion of the visceral form of acid sphingomyelinase deficiency (Niemann-Pick disease type B; NPB) or a much rarer disorder, variant adult visceral form of Niemann-Pick disease type C (NPC). To verify the histopathological findings, SMPD1, NPC1 and NPC2 genes were analysed. Two novel sequence variants, c.1997G>A (S666N) and c.2882A>G (N961S) were detected in the NPC1 gene. No pathogenic sequence variants were found either in the SMPD1 gene mutated in NPB or in NPC2 gene. The pathogenicity of both NPC1 variants was supported by their location in regions important for the protein function. Both variations were not found in more than 300 control alleles. Identified sequence variations confirm the diagnosis of the extremely rare adult visceral form of Niemann-Pick disease type C, which is otherwise dominated by neurovisceral symptoms. Although only three patients have been reported, this (most probably underdiagnosed) form of NPC should be considered in differential diagnosis of isolated hepatosplenomegaly with foam cells in adulthood.

A Faraday effect position sensor for interventional magnetic resonance imaging.

An optical sensor is presented which determines the position and one degree of orientation within a magnetic resonance tomograph. The sensor utilizes the Faraday effect to measure the local magnetic field, which is modulated by switching additional linear magnetic fields, the gradients. Existing methods for instrument localization during an interventional MR procedure often use electrically conducting structures at the instruments that can heat up excessively during MRI and are thus a significant danger for the patient. The proposed optical Faraday effect position sensor consists of non-magnetic and electrically non-conducting components only so that heating is avoided and the sensor could be applied safely even within the human body. With a non-magnetic prototype set-up, experiments were performed to demonstrate the possibility of measuring both the localization and the orientation in a magnetic resonance tomograph. In a 30 mT m(-1) gradient field, a localization uncertainty of 1.5 cm could be achieved.

Elevated and deregulated expression of HDAC3 in human astrocytic glial tumours.

Abnormal expression of histone deacetylases may contribute to the establishment of a cancer specific transcription profile. We examined expression of HDAC3 in human non-malignant gliosis and glial astrocytic tumours. Samples from four non-malignant gliosis and 17 astrocytic gliomas (six of grade II, one of grade III and ten of grade IV) removed for therapeutic purposes were assayed for HDAC3 expression at mRNA and protein levels. HDAC3 mRNA was detected in non-tumorous gliosis as well as in all examined glial tumours. Seven out of eleven examined high-grade tumours showed an elevated number of copies of HDAC3 mRNA. Western blot analysis detected high levels of expression of HDAC3 in the majority of the examined tumours. Immunohistochemistry and immunofluorescence made on a collection of 35 astrocytic tumours detected nuclear as well as cytoplasmic HDAC3 expression in all of those tumours. While the distribution of HDAC3 was both nuclear as well as cytoplasmic and moderate in intensity in non-malignant tissues and low-grade gliomas, high-grade tumours expressed HDAC3 in a focally deregulated pattern that included strongly pronounced cytoplasmic localization. Confocal microscopy and additional co-localization analysis detected nuclear HDAC3 in all tumours examined. We conclude that HDAC3 expression is elevated in human astrocytic tumours and its expression pattern is deregulated at the cellular level in high-grade gliomas.

Retrospective sequence analysis of the human PRNP gene from the formaldehyde-fixed paraffin-embedded tissues: report of two cases of Creutzfeldt-Jakob disease.

The definitive diagnosis of the CJD (Creutzfeldt-Jakob disease; very rare neurodegenerative disorder) can be established only on the basis of post-mortem examination of the central nervous system tissue. Formaldehyde-fixed paraffin-embedded (FFPE) tissue samples may thus constitute the only material available for molecular pathology analyses. We performed post-mortem analysis of the coding region of the prion-protein gene (PRNP)-sequence variations in two definite CJD cases suggestive of genetic form. Only FFPE tissues were available for molecular analyses. The PRNP gene open reading frame was amplified from the genomic DNA (FFPE isolated) in four overlapping, two round semi-nested PCR products that were directly sequenced. We found known pathogenic sequence variation g.532 G>A (Asp178Asn) in patient 1 but we did not find any pathogenic sequence variation in patient 2 despite her origin from the Slovak Orava region. Based on these results, we were able to discriminate between genetic and sporadic form of CJD in patient 1 and 2, respectively. The established method was found to be efficient for the sequence-variation analysis of the entire PRNP gene coding region using the genomic DNA isolated from the FFPE tissues; it can be employed in other retrospective molecular studies.

[Cardiac myxoma in the left atrium resulting in syncopes]. / Myxom levé síne jako prícina kolapsových stavu.

A case study of a 74-year-old woman with cardiac myxoma in the left atrium resulting in syncopes is discussed in this article.

Dependence between acute phase response, oxidative status and mastitis of cows.

There are many studies exploring the topic of acute phase response and oxidative status in inflammation of the mammary gland of cows. However, many phenomena are relatively not well known. Mastitis is associated with significantly higher concentrations of inflammatory and oxidative mediators in the cells and blood. Results of experiments have shown that there are evident changes in serum interleukins (IL), acidglycoprotein (alpha1AG), tumor necrosis factor (TNF), and haptoglobin (Hp). Thus, local as well as systemic inflammation might play important roles in increased mammary oxidative stress. Reactive oxygen species (ROS) have been implicated in the pathogenesis of a variety of diseases, including mastitis and in transgenic technology leading to production of new bacterial proteins, very important in prevention of mastitis. We can also observe an interaction between inflammatory and oxidative mediators. These results suggest an important role played by acute phase response and oxidative status in inflammation of the mammary gland.

Concentration of ascorbic acid in the blood of cows with subclinical mastitis.

The aim of the investigations was evaluation of ascorbic acia concentration in the blood or cows in the subclinical form of mastitis. The research was conducted on 56 cows. The cows were divided into 4 groups: A, B, and C with subclinical form of mastitis caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia col, and D control. The ascorbic acid concentration in the serum of cows was measured by the hydrazine method. We observed a marked decrease in ascorbic acid concentration in the serum of cows from experimental groups A, B and C, respectively: 34.2, 35.9, and 39.4 micromol/dm3 when we compared them with control group D--69.8 micromol/dm3. In cows with subclinical form of inflammation of the mammary gland, a decreased potential of antioxidant protection in the blood was noticed, which manifested itself as a lower ascorbic acid concentration.

The estimation of velocity distribution profile of Paramecium cytoplasmic streaming.

Cytoplasmic streaming of Paramecium bursaria and Paramecium tetraurelia was investigated by cinematographic techniques. Analysis of the records reveals the paraboidal character of the velocity distribution profiles in all arbitrarily chosen zones along the whole route of moving cytoplasm in the cell. According to the date obtained from cytoplasmic streaming analysis and food vacuole path, the geometry of the "channel" was described in terms of ellipsoid axes. Total volume changes of cytoplasm flowing through a given cross section in a given time unit were computed and no significant differences were found. The participation of a pressure gradient in motive force generation in cytoplasmic streaming is discussed.

Tricyclic antidepressant binding to lymphocyte membranes and changes during depression.

The binding of [3H]imipramine, its 2- and 4-nitroderivatives and [3H]desmethylimipramine to lymphocyte membranes was determined. IC50 values for drugs and neurotransmitters to inhibit [3H]imipramine binding to lymphocyte membranes were comparable with those for brain and thrombocyte membranes. The number of [3H]imipramine and [3H]desmethylimipramine binding sites increased in depressive patients, whereas the dissociation constants remained unchanged.

The effect of nickel compounds on immunophenotype and natural killer cell function of normal human lymphocytes.

In order to elucidate effects of nickel on human lymphocytes in vitro, peripheral blood mononuclear cells from normal donors were initially tested for viability in the presence of increasing concentrations of two selected nickel salts, sparingly-soluble nickel subsulfide (Ni3S2), and promptly-soluble nickel sulfate (NiSO4). After establishing the toxicity profile, the cells were cultured for 24 h with each compound at three nontoxic concentrations, 0.01 mM, 0.02 mM, and 0.04 mM, to determine its effect on lymphocyte immunophenotype and function. Cells were also cultured in the presence of 0.01-0.04 mM magnesium acetate, Mg(CH3COO)2, while still other cell samples were subjected to a mixture of Mg(CH3COO)2 plus either Ni3S2 or NiSO4 at equimolar concentration. Following the culture, the immunophenotype of the cells was determined by indirect immunofluorescence, using monoclonal antibodies to major differentiation antigens of peripheral blood mononuclear cells, and their natural killer activity toward K562 target cells was measured. Both nickel salts were found to exert distinct effects on lymphocyte phenotype. Exposure of cells to Ni3S2 resulted in the decline of CD4 and natural killer cell populations. NiSO4 diminished the abundance of natural killer cells and, to a limited extent, also of CD4 cells. The nickel salts tested suppressed natural cytotoxicity of peripheral blood mononuclear cells, with Ni3S2 acting more strongly than NiSO4. The addition of Mg(CH3COO)2 to a nickel salt during in vitro culture abolished the above inhibitory effects. Nickel and magnesium salts did not affect CD3, CD8, CD20, and CD11a cell populations. The results indicate that nickel salts have deleterious effects on human peripheral blood mononuclear cells in short-term in vitro culture, but the magnitude of these effects varies, depending on the cell subsets.