Pediatric Inflammatory Multisystem Syndrome (PIMS) - Potential role for cytokines such Is IL-6.

COVID-19 is a transmissible respiratory disease caused by coronavirus SARS-CoV-2, which is similar to SARS or MERS. Its increased severity was noted in aged patients usually over 65 years of age. Children and young people have an asymptomatic or mild course of the disease.Unfortunately, the number of children with problems after mild or asymptomatic COVID-19 recovery is increasing and their troubles resemble Kawasaki disease, although the laboratory findings seem to be different. This condition is called pediatric inflammatory multisystem syndrome (PIMS), and it is a new disease seen in children directly influenced by previous SARS-CoV-2 infection. The literature reports that PIMS typically follows 2-4 weeks after SARS-CoV-2 infection. The clinical symptoms of the affected children are extremely complex, ranging from gastrointestinal to cardiovascular problems with frequent skin and mucosal manifestations, and without intensive treatment they can be fatal. The exact causes of PIMS are recently unknown, however, it is explained as hyperactivation of immunity.In this minireview, we summarize data on the prominent role of the IL-6-IL-6R-STAT3 axis in PIMS aetiopathogenesis. Therapeutic manipulation of IL-6 or IL-6 receptor could be an approach to the treatment of children with severe PIMS.

[Isolated Perfusion of the Upper Extremity with TNF-α - Double Venous Cannulation]. / Izolovaná perfuze horní koncetiny s TNF-α - dvojitá kanylace zilního systému.

BACKGROUND: The authors present a technical variation of the standard cannulation for cardiopulmonary bypass perfusion during hyperthermic isolated limb perfusion (ILP) procedures in selected patients with unresectable soft tissue sarcoma or malignant melanoma. PATIENTS: Of 55 ILP procedures performed at our institution since the procedure was established in 2009, nine were performed at the upper extremity. Standard single venous cannulation was used in five cases, and extended, double venous cannulation in the last four. The standard technique for brachial vein cannulation in a small compartment of the upper extremity entails a problematic and longer perfusion of the upper extremity. This is due to the lower flow rate in the venous system and relatively large surface area with respect to weight. We present a simple technique based on a "Y" cannulation of the venous system via the deep brachial vein and superficial venous system via the basilic vein, delivering a 20% increase in flow rate in the extracorporeal circulation. Faster heating of the upper extremity and a stable thermal environment throughout upper-extremity ILP are essential for successful treatment. CONCLUSION: Extended technique of venous cannulation for extracorporeal circulation setting, due to their advantages, became standard in the upper limb ILP procedure at our institution.Key words: isolated limb perfusion - malignant melanoma - soft tissue sarcoma - upper limb - extracorporeal circulation The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 8. 1. 2017Accepted: 15. 1. 2017.

Phylogeny, regeneration, ageing and cancer: role of microenvironment and possibility of its therapeutic manipulation.

Data about the possible correlation between reduction of the regeneration capacity in the course of phylogeny and formation of malignant tumours have been summarized from invertebrates to mammals. The evolutionarily increasing complexity of body building plane and expectancy of longevity in the course of phylogeny seems to be grossly negatively correlated with diminished regeneration capacity, but positively with increased occurrence of malignant tumours. A certain evolution-based switch-off mechanism reducing the extent of regeneration in developmentally complicated and long-living animals such as mammals and birds can be hypothesized and benefits of loss of this ability are discussed. This high incidence of malignancies seems to be related, in addition to other factors, to prolonged and cumulative exposure to cancerogenic stimuli in the course of lifetime. Longevity, supported by the progress and availability of medical care to the population, has been unveiling this phenomenon during recent decades. From this point of view, ageing represents the main risk for cancer acquisition. The probable role of microenvironment in all the discussed phenomena such as healing/regeneration, inflammation, and cancer is discussed and targeting of microenvironment is consequently predicted as a possible therapeutic target where controlled manipulation may represent a new approach to the treatment of cancer patients.

Fibroblasts prepared from different types of malignant tumors stimulate expression of luminal marker keratin 8 in the EM-G3 breast cancer cell line.

It is widely recognized that stromal fibroblasts significantly influence biological properties of multiple tumors including breast cancer. However, these epithelial-mesenchymal interactions seem to be essential in tumor biology and it is not fully clear whether this interaction is tumor type-specific or has a more general non-specific character. To elucidate this question, we tested the effect of cancer-associated fibroblasts (CAFs) isolated from different types of tumors (breast cancer skin metastasis, cutaneous basal cell carcinoma and melanoma, squamous cell carcinoma arising from oral cavity mucous membrane) on the EM-G3 breast cancer cell line. The results were compared with control experiments using normal human dermal fibroblasts, 3T3 mouse fibroblasts, and 3T3 fibroblasts influenced by the fibroblasts prepared from the basal cell carcinoma. Our results demonstrated that expression of luminal marker keratin 8 was influenced only by CAFs prepared from any tested tumors. In contrast, all tested types of fibroblasts showed a strong stimulatory effect on the expression of basal/myoepithelial marker keratin 14. The CAFs also elevated the number of cells with positivity for both keratins 8 and 14 that are similar to ductal originated precursor cells. The expression of proliferation marker Ki67 was not influenced by any of the tested fibroblasts. In conclusion, our data indicate that CAFs are able to influence the phenotype of a breast cancer cell line and this effect is based on a tumor type-unspecific mechanism. Finally, a clear functional difference between normal and CAFs was demonstrated.

Comparative analysis of the nuclear presence of adhesion/growth-regulatory galectins and reactivity in the nuclei of interphasic and mitotic cells.

Nuclear galectins participate in splicing of pre-mRNA. In this study we detected galectins-1, -2, -3 and -7 and their glycoligands in three types of cells: fibroblasts, cancer epithelial cells and melanoma cells. The results demonstrated that the nuclear expression of distinct types of galectins and their ligands in interphasic nuclei is dependent on the cell type. The extensive binding of labelled galectins-1 and -2 to mitotic cells (around chromosomes, in mitotic spindle and in bridge connecting both daughter cells) suggests their role during the cell division.

[Cytostatic hyperthermic isolated limb perfusion (HILP) in VFN (General Faculty Hospital in Prague)]. / Cytostatická hypertermická perfuze izolované koncetiny (HILP) ve VFN.

Hyperthermic isolated limb perfusion (HILP) is a standardized method of treatment in selected patients with in-transient locoregional recurrence/methastasis of melanoma or, some other soft tissue tumors (incl. sarcoma etc.) Authors present history and current status of this treatment modality in General University Hospital in Prague. During one year period (7/2009-6/2010) 10 patients were indicated for this procedure. We performed 13 procedures (3x redo), 11 in lower extremity and 2 in upper extremity. There was no serious complication in this cohort of patiens. Multidisciplinar approach is indicated in melanoma patients care.

Two-year follow-up of two patients after severe thallium intoxication.

Information on the prognosis and electrophysiological follow-up of severe thallium poisoning is limited. We report two patients (mother and daughter) who were repeatedly exposed to thallium poisoning experienced hair loss, polyneuropathy, and visual impairment. Nerve conduction studies (NCSs), visual evoked potentials (VEP), brainstem auditory evoked potentials (BAEP) changes, and optical neuropathy developed within a few months latency after the first subjective signs. Normal findings of these electrophysiological methods in the first 2 weeks therefore led in one of our patients to exclusion of thallium as the cause of symptoms. Thallium poisoning was, however, later confirmed by toxicological analysis of blood and/or urine and feces in both the patients and in the microscopic hair analysis of the daughter. Both patients were treated with Prussian blue that increased the elimination of thallium in urine and feces. The hair loss was fully reversible. During a 2-year follow-up after the poisoning, polyneuropathy in the lower extremities improved substantially, but residual impairment in both motor and sensory function, NCSs, VEP, and BAEP remained. Additionally, severe asymmetrical vision impairment persists in both women, with central scotomata and impaired color discrimination in both eyes. Substantial improvement of their visual function is unlikely.

Immunohistochemical fingerprinting of the network of seven adhesion/growth-regulatory lectins in human skin and detection of distinct tumour-associated alterations.

Glycans of natural glycoconjugates are considered as a source of biological information relevant to cell adhesion or growth. Sugar-based messages are decoded and translated into responses by endogenous lectins. This mechanism assigns a functional dimension to tumour-associated changes of glycosylation. Consequently, it calls for mapping the lectin presence in tumours. Such an analysis has so far commonly been performed with the scope to determine expression of a few distinct proteins, e.g. from the effector family of galectins with focus on galectins-1 and -3. Due to the emerging evidence for functional divergence among galectins it is timely to address the challenge to evaluate their presence beyond these few family members. Having raised a panel of non-cross- -reactive antibodies against seven human galectins covering all three subfamilies, we de scribe their expression profiles in human skin. Comparison of normal and malignant tissues enabled us to define galectin-type-dependent alterations, arguing in favour of distinct functionalities. It is concluded that comprehensive monitoring performed to define the different aspects of the galectin network, as documented in this pilot study, is advisable for future histopathologic studies aimed at delineating clinical correlations.

Phenotypic characterization of human keratinocytes in coculture reveals differential effects of fibroblasts from benign fibrous histiocytoma (dermatofibroma) as compared to cells from its malignant form and to normal fibroblasts.

BACKGROUND: Benign and malignant fibrous histiocytoma present with a considerable difference concerning cellular organization in their vicinity. OBJECTIVE: Normally appearing epithelium covers the malignant form in contrast to hyperplastic epidermis for benign tumors. It is an open question as to whether the tumor-associated fibroblasts are capable to affect phenotypic features of normal keratinocytes, prompting this comparative analysis. METHODS: Fibroblasts were isolated from benign and malignant fibrous histiocytomas, respectively, and also from normal dermis. The resulting cell populations were thoroughly characterized immunocytochemically using a large panel of antibodies. The three fibroblast preparations were cocultured with normal interfollicular keratinocytes. Their phenotype was characterized for distinct properties including differentiation and proliferation. RESULTS: Fibroblasts prepared from both tumor types were phenotypically practically identical with normal dermal fibroblasts. Their activities on keratinocytes were different. Cells prepared from benign fibrous histiocytoma were capable to effect strong expression of keratin 19 and production of a galectin-1-rich extracellular matrix. Fibroblasts isolated from malignant fibrous histiocytoma led to a phenotype very similar to that when keratinocytes were cocultured with normal dermal fibroblasts. CONCLUSION: Fibroblasts prepared from benign fibrous histiocytoma were biologically active on keratinocytes in a particular manner. Our results on fibroblast activity are suggested to be relevant for morphologic differences observed in vivo between normal epidermis and epidermis adjacent to the studied tumor types.

Differential regulation of galectin expression/reactivity during wound healing in porcine skin and in cultures of epidermal cells with functional impact on migration.

The glycophenotyping of mammalian cells with plant lectins maps aspects of the glycomic profile and disease-associated alterations. A salient step toward delineating their functional dimension is the detection of endogenous lectins. They can translate sugar-encoded changes into cellular responses. Among them, the members of the lectin family of galectins are emerging regulators of cell adhesion, migration and proliferation. Focusing on galectins-1, -3 and -7, we addressed the issue whether their expression is regulated during wound healing in porcine skin as model. A conspicuous upregulation is detected for galectin-1 in the dermis and a neoexpression in the epidermis, where an increased level of galectin-7 was also found. Applying biotinylated tissue lectins as probes, the signal intensities for accessible binding sites decreased, intimating an interaction of the cell lectin with reactive sites. In contrast, galectin-3 parameters remained rather constant. Of note, epidermal cells in culture also showed an increase in expression/presence of galectin-1, measured on the levels of mRNA and protein, in this case by Western blotting and quantitative immunocytochemistry. Used as matrix, galectin-1 conferred resistance to trypsin treatment to attached human keratinocytes and reduced migration into scratch-wound areas in vitro. This report thus presents new information on endogenous lectins in wound healing and differential regulation among the three tested cases.

Galectin-7: will the lectin's activity establish clinical correlations in head and neck squamous cell and basal cell carcinomas?

The human lectin galectin-7 (Gal-7; p53-induced gene-1) has anti- and pro-malignant features in different in vitro models. We tried to clarify relation of its expression to cellular and clinical parameters in head and neck squamous and basal cell carcinomas. Using a non-cross-reactive antibody, immunohistochemical staining in squamous cell epithelia (epidermis, epithelium of oropharynx and larynx) (n = 57), squamous cell carcinomas (n = 47) and lymph node metastases (n = 25), as well as basal cell carcinomas (n = 10) were studied. This monitoring was flanked by processing to assess the level of differentiation (cytokeratins 10 and 14), proliferation (Ki67) and basal lamina formation (collagen IV). The results were correlated with clinical and pathological findings (grading, TNM-staging, extracapsular spread, angio- and lymphangioinvasion, perineural invasion, recurrence and survival). Gal-7 resides in all layers of epithelia with cytoplasmic and nuclear localization in normal specimens. Basal cell carcinomas were devoid of the Gal-7 respective signal. Squamous cell carcinomas were positive, presenting different staining profiles. Intense staining was predominantly found in squamous cell cancers with high degrees of differentiation and keratinization. Fittingly, poor level of differentiation (P = 0.0009), absence of keratinization (P = 0.0105) and significant discontinuity or absence of collagen IV expression in the peritumoral basal lamina (P = 0.0024) was found in Gal-7-negative tumors. Gal-7 presence was not related to gender, primary tumor site, T-stage, N-stage, clinical stage, extracapsular spread, angio- and lymphangioinvasion, perineural spread or treatment outcome at a statistically significant level. Immunohistochemical analysis revealed a positive correlation for differentiation and keratinization to Gal-7 presence in squamous cell carcinomas. Absence of Gal-7 expression was detected in basal cell carcinomas. These clinical data delineate Gal-7 influence on differentiation in vivo, without evidence for a role in dissemination reported for lymphoma.

[Galectins in squamous cell carcinomas of the head and neck cancers]. / Galektiny v dlazdicových karcinomech hlavy a krku.

Cancers of head and neck represents about 5% of all tumors. 80 to 90% of these tumors are constituted of squamous cell carcinomas. Despite a rapid progress in diagnostics and therapy the overall 5-year survival of this type of cancer is among the lowest of the major cancer types. This unfavourable situation needs the extensive research to found new markers to better characterize biological behavior of tumors as a rational background for more sophisticated therapeutic modalities. Among the most promising markers are endogenous lectins called galectins and their ligands. Especially galectin-1, -3 and -7 play a key role in pathology of squamous cell carcinomas.

Human galectin-2: nuclear presence in vitro and its modulation by quiescence/stress factors.

Galectins have the particular capacity to interact with distinct proteins, in addition to the typical reactivity of lectins with glycans. Therefore, they can be functionally active when residing at places other than the membrane or extracellular matrix. In fact, nuclear presence of galectins-1 and -3 is solidly documented but it is an open question whether these two cases are exceptional within this lectin family. Thus, galectin-2, which shares 43% sequence identity on the protein level with galectin-1, warrants study in this respect. Based on initial immunohistochemical evidence we herein address the issue as to whether this galectin can join the category of nuclear lectins. To do so we studied different types of cell in vitro using an antibody preparation free of cross-reactivity against other tested galectins. The immunocytochemical experiments revealed that galectin-2 was present in nuclei of murine 3T3 fibroblasts and also genetically engineered human colon carcinoma cells with stable ectopic expression. Transport of galectin-2 to the nucleus could be enhanced by physical (UV light), chemical (mitomycin C, serum withdrawal) or cell biological (coculture with stromal cells) treatment modalities. As a means of further characterizing the staining profile cytochemically, a series of markers with well-defined site of residency within the nuclear compartment was tested in parallel. Importantly, no colocalization with galectins-1 and -3 and the splicing factor SC35 was detectable, the former cases also serving as inherent specificity control. In contrast, a similarity was uncovered in the case of the promyelocytic leukemia (PML) protein as marker of PML nuclear bodies. In aggregate, nuclear localization is documented for galectin-2. This attribute should thus not be considered as an exceptional finding confined to galectins-1 and -3. That even closely related family members, here galectins-1 and -2, exhibit distinct intranuclear localization patterns gives ensuing research a clear direction.

Stromal fibroblasts from basal cell carcinoma affect phenotype of normal keratinocytes.

BACKGROUND: Epithelial-mesenchymal interactions are important not only to direct the course of prenatal development of skin and its appendages but also to influence the behaviour of transformed epithelial cells. OBJECTIVES: Evaluation of the role of stromal fibroblasts on the phenotype of epithelial cells of basal cell carcinoma (BCC). METHODS: The phenotype of human BCC was compared with the in vitro model where the growth and phenotypic pattern of normal human keratinocytes were monitored in co-culture with fibroblasts prepared from stroma of BCC (BCCFs), with normal dermal fibroblasts or with two established fibroblast lines. We visualized the expression of a panel of keratins, three types of endogenous lectin [galectin (Gal)-1, Gal-3 and Gal-7], binding sites for Gal-1 and Gal-3, a proliferation marker Ki67, nucleolar protein nucleostemin (NuclS) and membrane protein Ber-EP4. A phenotype and karyotype of BCCFs were also monitored. BCCFs were also grafted to NOD/LtSz-Rag1(null) mice to evaluate their malignant potential. RESULTS: Prolonged cultivation of BCCFs has led to morphological changes, loss of contact inhibition, loss of fibroblast surface antigens and progressive aneuploidity. However, a fully malignant phenotype did not develop as these cells did not form tumours in immunodeficient mice. Co-culture of BCCFs with normal keratinocytes in vitro led to their phenotypical changes resembling those in BCC, namely, expression of keratin 19. These keratinocytes also strongly express nuclear binding sites for Gal-1 and NuclS. This phenotype was not observed when normal keratinocytes were cultured with nontumour-originated fibroblasts. CONCLUSIONS: These observations indicate that BCCFs may differ from normal fibroblasts and may play a regulatory role in BCC biology.

Glycophenotype of psoriatic skin.

Psoriasis is considered an auto-immune disease with consequential keratinocyte hyperproliferation resulting in specific architecture of psoriatic skin. This process is associated with phenotypical keratinocyte changes including an altered carbohydrate expression pattern studied by labelled plant lectins. Expression of endogenous lectins and their reactive glycoligands are differentiation-dependent in squamous epithelia including epidermis. However, no data are available on psoriatic skin, although this disease represents an important medical problem. We investigated the expression of galectin-1, -3, -7 and the presence of their glycoligands in the psoriatic skin and compared the results with the normal skin samples. The results were correlated to expression patterns of cytokeratin 10 and cytokeratin peptide 37 as markers of keratinocyte differentiation as well as to the expression of proliferation marker Ki67. Contrary to normal epidermis, the psoriatic epithelium expressed no galectin-3 and no glycoligands for galectin-1. Strong expression of galectin-3/galectin-3-reactive glycoligands in capillaries of psoriatic dermis represents one of the most important findings demonstrating the activation of endothelium in the course of the disease. The keratin expression pattern was not affected in psoriatic skin compared with normal epidermis. In conclusion, the altered galectin expression and binding pattern in psoriatic skin indicates the modified process of keratinocyte maturation in hyperactivated psoriatic epithelium. The enhanced expression of galectin-3/galectin-3-reactive glycoligands in dermal capillaries of psoriatic skin can be important for rearrangement of the capillary network and migration of inflammatory cells to psoriatic skin.

Validation of a self-administered questionnaire to screen for prenatal alcohol use in Northern Plains Indian women.

OBJECTIVE: This study among American Indian prenatal patients was conducted to validate a self-administered questionnaire (SAQ) designed to (1) identify women who had consumed alcohol during pregnancy, (2) identify women who may be at risk of drinking during pregnancy, and (3) determine the quantity and frequency of alcohol and other substance use just before and during pregnancy. METHODS: The validation involved three components: (1) review of the SAQ responses by a public health nurse; (2) structured patient interview with the research nurse; and (3) medical record abstraction postpartum. RESULTS: Compared to extensive interview and medical record data, the SAQ is sensitive (76.6%) and specific (92.8%) in detecting pregnant women who had consumed alcohol during pregnancy. CONCLUSION: The SAQ is a useful screening tool for alcohol use in this population.

Relationship of prenatal alcohol use with maternal and prenatal factors in American Indian women.

Demographic factors and patterns of substance use among women who did not consume alcohol during pregnancy were compared to women who did consume alcohol during pregnancy. One-hundred seventy-seven Northern Plains Indian women who received prenatal care at an urban clinic in a rural state were screened for substance use as part of the validation study with a self-administered questionnaire. Women who drank during pregnancy were more likely to be single and have less education than women who did not drink. While most of the women in the study had available transportation resources, the women who drank during pregnancy were less likely to have transportation than the women who did not drink. Women who drank during pregnancy consumed more alcohol more frequently before pregnancy than did women who drank before but not during pregnancy. Compared to women who did not drink during pregnancy, women who drank during pregnancy were more likely to smoke cigarettes and use illicit drugs, to have parents who drank, to feel they drank the same or more than other pregnant women, or to have experienced more relationship breakups and physical and emotional abuse. Prenatal patients who drink alcohol during pregnancy need more intensive counseling regarding their multiple risk behaviors.