Clinical analysis of tethered cord syndrome and gene expression profiling of wound healing surgery.

Introduction: The method to prevent progression of symptoms in tethered cord syndrome (TCS) is neurosurgery. However, postoperative wound healing is a lengthy process and is hindered by the release of cerebrospinal fluid (CSF) through the wound. To the best of the authors' knowledge, there is no study evaluating the changes in the expression of factors involved in the wound healing process after neurosurgery for TCS. Aim: To clinically analyse 2 cases of TCS and evaluate the change in expression of selected genes during the postoperative wound healing process. Material and methods: Determination of TCS in two adult patients (woman, aged 26 years; man, aged 53 years) was based on magnetic resonance imaging (MRI). After confirming the initial diagnosis, a neurosurgical procedure was performed to remove the intrathecal spreading adipoma and transect the medullary terminal thread in patients. In the postoperative period, impaired wound healing was noted as a result of CSF secretion through the surgical wound. Results: Molecularly, there was an increase in expression of all genes assessed in skin biopsy specimens compared to skin samples. Impaired postoperative wound healing after neurosurgery for TCS is expected due to CSF leakage through the surgical wound. The greatest changes were noted for metalloproteinases (MMPs) and four isoforms (A-D) of vascular endothelial growth factor A-D (VEGF-A-D; p < 0.05). Conclusions: Changes in the expression of our selected genes can be used to monitor and predict the process of wound healing and scar formation, which occurred in our cases at 19 and 20 days after surgery.

Polyspikes and Rhythmic Polyspikes During Volatile Induction of General Anesthesia With Sevoflurane Result in Bispectral Index Variations.

BACKGROUND: Although electroencephalography (EEG)-based indices may show artifactual values, raw EEG signal is seldom used to monitor the depth of volatile induction of general anesthesia (VIGA). The current analysis aimed to identify whether bispectral index (BIS) variations reliably reflect the actual depth of general anesthesia during presence of different types of epileptiform patterns (EPs) in EEGs during induction of general anesthesia. METHODS: Sixty patients receiving either VIGA with sevoflurane using increasing concentrations (group VIMA) or vital capacity (group VCRII) technique or intravenous single dose of propofol (group PROP) were included. Monitoring included facial electromyography (fEMG), fraction of inspired sevoflurane (FiAA), fraction of expired sevoflurane (FeAA), minimal alveolar concentration (MAC) of sevoflurane, BIS, standard EEG, and hemodynamic parameters. RESULTS: In the PROP group no EPs were observed. During different stages of VIGA with sevoflurane in the VIMA and VCRII groups, presence of polyspikes and rhythmic polyspikes in patients' EEGs resulted in artifactual BIS values indicating a false awareness/wakefulness from anesthesia, despite no concomitant change of FiAA, FeAA, and MAC of sevoflurane. Periodic epileptiform discharges did not result in aberrant BIS values. CONCLUSION: Our results suggest that raw EEG correlate it with values of BIS, FiAA, FeAA, and MAC of sevoflurane during VIGA. It seems that because artifactual BIS values indicating false awareness/wakefulness as a result of presence of polyspikes and rhythmic polyspikes in patients' EEGs may be misleading to an anesthesiologist, leading to unintentional administration of toxic concentration of sevoflurane in ventilation gas.

Trichopsychodermatology: trichotillomania and trichophagia leading to Rapunzel syndrome.

Introduction: The Rapunzel syndrome occurs when the trichobezoar (hair ball) extends beyond the small intestine and sometimes even into the colon, producing long, tail-like hair extensions. Aim: To present cases of trichobezoars, an extremely rare human intestinal disease caused by the ingestion of hair (trichophagia). Material and methods: In this retrospective study, we assessed the medical records of 2 patients diagnosed with Rapunzel syndrome admitted to Academic Clinical Hospital No. 2 in Rzeszow, Poland. Results: The first patient was a 15-year-old girl. The abdominal ultrasound examination revealed an abnormal, non-compressible structure with the approximate dimensions of 12 × 11 cm in the epigastrium, with a strong shadow obscuring the view. Gastroscopy was performed and a tumour sample was taken for histopathological examination, which confirmed the diagnosis of a trichobezoar. The patient's mother did not agree to her daughter's psychiatric treatment. The second patient was a 15-year-old girl who reported to the Emergency Room in critical condition due to dehydration and long-term emesis with symptoms of cachexia. Ileotomy with the removal of two trichobezoars with a diameter of about 5 cm and 7 cm was performed. The patient was discharged for treatment at the Mental Health Clinic for trichotillomania. Conclusions: Trichobezoars give non-specific symptoms that may imitate other diseases for example tumours. Psychotherapy is the recommended treatment and follow-up visits are important in preventing relapses.

Cutaneous metastasis as the first sign of renal cell carcinoma - crossroad between literature analysis and own observations.

Introduction: Metastatic disease can be observed in nearly 25% of all renal cell carcinoma cases (RCC). This can present in the skin as a late symptom of the disease or as a manifestation of an undetected, asymptomatic RCC. Aim: To review the literature and present a case study of cutaneous metastasis in primary genitourinary malignancy, especially RCC in order to broaden the related knowledge. Material and methods: The first stage of our work was focused on presenting the results of the literature review about cutaneous metastasis in the patients with renal cell carcinoma. Next, taking into account that this type of metastasis associated with RCC is relatively rare, we have decided to present a patient aged 68 with incidentally detected skin metastases of RCC located on the scalp. Results: It was diagnosed as a primary manifestation of advanced disease. Computed tomography scans showed a solid mass in the left kidney. Following surgical excision of the skin lesion, a left-side nephrectomy was performed. Conclusions: Cutaneous metastases of RCC present an unfavourable prognosis, however, remission is possible subsequent to early diagnosis and appropriate surgical excision.

Microbiological effects in patients with leg ulcers and diabetic foot treated with Lucilia sericata larvae.

Lucilia sericata bottle fly worms can be used to heal infected, chronic, or necrotic wounds, including those associated with ulceration and diabetic foot. The study aimed to evaluate changes in the microflora in patients treated with L sericata larvae due to leg ulcers and diabetic foot. One hundred twenty-nine patients diagnosed with lower limb ulceration and diabetic foot were enrolled in the study, of which 80 of them met the eligibility criteria for maggot debridement therapy (MDT). On the contrary, 49 unqualified patients were offered ozone therapy (22 with leg ulcers; 27 with diabetic foot). In each of these patients, a microbiological swab was performed before and after the start of therapy. The group of 80 patients was further divided into four equal groups in terms of the treated area (lower leg vs foot) and the number of larvae/cm2 (5 vs 10). Twenty-three particular species of bacteria in the infected wound were studied microbiologically in terms of presence/absence within the wound environment before and after treatment of patients with diabetic foot and lower limb ulceration. It was noted that there was a more intensive bacterial accumulation in the feet of patients compared to legs; furthermore, this applies to almost all analysed species. Diabetes status is also a clinical factor that generates a lower chance of bacterial appearance in the wound environment. Densification of MDT larvae per wound area unit also reduced the chance of the presence of Corynebacterium species, Enterobacteriaceae, Pseudomonas aeruginosa, Staphylococcus aureus MSSA, and Streptococcus coagulase negativa; however, it increased the likelihood of occurrence for Proteus mirabilis and the Proteus species. A microbiological analysis in this non-reference study shows the efficacy of larval therapy for leg and foot ulcers. Rearrangement of the microflora within the wound has been reported as a result of the therapy.

Ocular Surface Preparation Before Keratoprosthesis Implantation.

INTRODUCTION: This study aimed to evaluate the surgical treatment results for conjunctival limbal autograft (CLAU) and keratolimbal allograft (KLAL) in various types of limbal stem cell deficiency (LSCD) etiologies performed in order to achieve a stable ocular surface prior to KPro implantation. METHODS: We analyzed the outcomes of the surgical treatment of 43 eyes of 39 patients with LSCD as an initial treatment preparing patients' ocular surface for KPro implantation. The most common causes were ocular trauma (50.7%), mainly alkali burns (77%); autoimmune causes, mainly ocular cicatricial pemphigoid (OCP; 17.4%); infection (15.9%) including Lyell's syndrome/Stevens-Johnson syndrome (LS/SJS; 16%). In all 17 eyes operated on with CLAU, this procedure was performed once. Similarly, one uncomplicated KLAL procedure in one eye was performed in 10 women and 19 men. In another one woman and three men, KLAL was performed in both eyes. In one man with Lyell's syndrome, the KLAL operation was performed three times in one eye. Follow-up was at least 12 months. RESULTS: Visual acuity (VA) improved in 17 eyes (31%) and remained unchanged in 38 eyes (69%). VA improved from light perception to hand movements in three eyes (16%) from the CLAU group of patients and eight eyes (15%) from the KLAL group; VA improved from hand movements to finger counting in two eyes (12%) post CLAU and two eyes (4%) post KLAL operation. The most common complication of surgical treatment was persistent epithelial defect that was refractory to medical treatment in 32 eyes (58%), 5 eyes post CLAU and 27 post KLAL. Corneal conjunctivalization (19%) and neovascularization (29%) were present on the corneal edge of the graft. Symblephara recurred within 3 months in nine eyes (17.3%) after KLAL, including four eyes that had been chemically burned and five eyes with LS/SJS. DISCUSSION: Pretreatment with CLAU or KLAL procedures in severely damaged ocular surfaces allows the ocular surface to be prepared for safe KPro implantation with sufficient tissue surroundings with less conjunctivalization and deeper conjunctival fornices.

Analysis of the influence of adalimumab to the expression pattern of mRNA and protein of TGF-ß1-3 in dermal fibroblast exposed to lipopolysaccharide.

INTRODUCTION: Psoriasis is a inflammatory illness, where incorrect expression of cytokines and bacteria lipopolysaccharide are observed. In the therapy of moderate to severe psoriasis anti-TNF drugs, i.e. adalimumab are used which have the influence for secreting another cytokines, such as transforming growth factor-ß (TGF-ß). AIM: To analyse the expression profile of mRNA TGF-ß1-3 and proteins (TGF-ß1 and TGF-ß2) it codes in normal human dermal fibroblasts (NHDF) exposed to bacterial lipopolysaccharide (induction of inflammation) and adalimumab (anti-TNF drug). MATERIAL AND METHODS: NHDFs treated with bacterial lipopolysaccharide at a medium concentration of 1 µg/ml for 8 h, and then added to an adalimumab culture at a concentration of 8 µg/ml and continued exposure of the fibroblasts to it for 2, 8 and 24 h. The molecular analysis included microarray, RTqPCR and ELISA assays. RESULTS: Treating the skin fibroblast cells with LPS resulted in significant statistical changes in the expression of TGF-ß1 (↑) and TGF-ß2 (↓) in comparison to the control culture. Likewise, after adding adalimumab to the culture of NHDF treated previously with LPS, significant changes in the expression of TGF-ß1 (↑) and TGF-ß2 (↓) were noted in comparison to the control culture (p < 0.05). On the protein level it can be determined that LPS and adalimumab cause an increase in the concentration of TGF-ß1 and a decrease in the expression of TGF-ß2 in comparison to the control culture. CONCLUSIONS: Blocking the signalling dependant on TNF-α using adalimumab causes an increase in the expression of TGF-ß1 and a simultaneous decrease in the case of TGF-ß2.

Emergency Medical Teams Interventions due to Cardiovascular Diseases in 2018: Polish Regional Observational Study.

Background and objectives: The goal of this work was to assess the interventions for cardiovascular causes (ICD-10: I) and analyze the time between the request for intervention and the arrival of the Medical Emergency Team realized by the Voivodeship Rescue Service in Katowice in the period between 1 January 2018 to 31 December 2018. Materials and Methods: Analysis of the characteristics of the interventions was completed based on the information contained on the dispatch order cards and medical emergency services. Statistical analysis was done using the Chi-square test (p < 0.05). Results: Out of 211,548 cases, 26,672 were associated with cardiovascular diseases. It can be observed that the large majority of interventions took place in urban areas (89.98%; 23,998 cases), whereas only 11.02% took place in rural areas (2674 cases). The most common cause for medical interventions being made by the Medical Emergency Team was primary hypertension-11,649 cases. The average arrival time to urban areas was 9 min and 12 s ± 3 min and 54 s, whereas for rural areas it was 11 min and 57 s ± 4 min and 32 s (p < 0.05). Conclusions: It can be observed that the Medical Emergency System in Katowice operates accordingly with the intentions of the legislator. The obtained data also indicates that there is a high societal awareness of the residents about the purpose of the Medical Emergency Team.

Salinomycin Modulates the Expression of mRNAs and miRNAs Related to Stemness in Endometrial Cancer.

BACKGROUND: Salinomycin, an ionophore antibiotic, has a strong anti-cancer effect, inducing the apoptosis of cancer cells and cancer stem cells. OBJECTIVE: The aim of the study was to assess the influence of salinomycin on the expression profile of genes related to stemness and miRNA regulating their expression in endometrial cancer cells. METHODS: Endometrial cancer cells of cell line Ishikawa were exposed to salinomycin at concentrations in the range of 0.1-100 µM, with the aim of determining its pro-apoptotic potential and the concentration which would cause the death of 50% of the cells (Sulforhodamine B test). In the following stages, the cells were incubated with the drug at a concentration of 1µM for 12,24 and 48 hour periods and compared to the control. Determining the changes in the expression of the genes related to stemness and regulating their miRNA was done using the microarray technique and RTqPCR. ELISA assay was performed in order to determine the level of TGFß2, COL14A1, CDH2, WNT5A in cell culture under salinomycin treatment in comparison to the control. RESULTS: Salinomycin caused the apoptosis of cells. For the concentration of 0.1 µM, a decrease in the population of living cells by 11.9% was determined. For 1 µM, it was 49.8%, for 10 µM -69.4%, and for a concentration of 100 µM - 87.9%. The most noticeable changes in the expression caused by the addition of salinomycin into the culture were noted for mRNA: TGFß2; WNT5A (up-regulated); COL14A1; CDH2 (down-regulated), as well as miRNA: hsa-miR-411 (up-regulated); hsa-miR-200a; hsa-miR-33a; hsa-miR-199a; hsa-miR-371-5p; hsa-miR-374; hsa-miR-374b (down-regulated). CONCLUSION: It was confirmed that salinomycin has an influence on the stemness process. The most noticeable changes in the expression were noted for mRNA: TGFß2; COL14A1; CDH2; WNT5A, as well as for miRNA: hsa-miR-200a; hsa-miR-33a; hsa-miR-199a; hsa-miR-371-5p; hsa-miR-411; hsa-miR- 374a; hsa-miR-374b.

Evaluation of Variances in VEGF-A-D and VEGFR-1-3 Expression in the Ishikawa Endometrial Cancer Cell Line Treated with Salinomycin and Anti-Angiogenic/Lymphangiogenic Effect.

BACKGROUND: In cancer, an excessive and uncontrolled process of creating new blood and lymphatic vessels that play a key role in the metastasis process can be observed. The Vascular Endothelial Growth Factor (VEGF-A,-B,-C,-D) family together with their specific receptors (VEGFR-1,-2,- 3) plays a key role in these processes, therefore, it would be reasonable to determine the correct pattern of their expression. OBJECTIVES: The study aimed to assess the use of salinomycin as an anti-angiogenic and anti-lymphangiogenic drug during endometrial cancer by examining changes in the expression pattern of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGFR-1, VEGFR-2 and VEGFR-3 depending on the treatment period of the Ishikawa endometrial cancer cells with salinomycin in comparison to the control culture. MATERIALS AND METHODS: To determine how influential salinomycin was on the expression of both mRNAs, 1 µM of the drug was added to the cell culture and then it was cultured all together for 12, 24 and 48 hour periods. The cells that made up the control culture were not treated with salinomycin. To determine the changes in the expression profile of the selected genes, we used the microarray, techniques: RTqPCR and ELISA (p<0.05). RESULTS: For all isoforms of VEGF-A-D as well as receptors of VEGFR-1-3, a decrease in expression under the influence of salinomycin was noted. For VEGF-A and VEGFR-1, the difference in the expression between the culture treated with salinomycin in comparison to the control was statistically significant (p=0.0004). In turn, for VEGF-B, the difference between the culture exposed for 24 hours in comparison to the control (p=0.00000) as well as the comparison between H48 vs. C (p=0.00000) was statistically significant. In reference to VEGF-C, VEGFR-2 and VEGFR-3, the statistical analysis showed the significant difference in expression between the culture incubated with the drug for 12, 24 and 48 hours in comparison to the control as well as between the selected times. For all of these comparisons, p=0.00000 was utilized. CONCLUSION: Salinomycin changes the expression pattern of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGFR-1, VEGFR-2, and VEGFR-3 in endometrial cancer cells. The obtained results suggest that salinomycin might exert the effect via VEGF signaling pathways.

In silico analysis of CpG islands and miRNAs potentially regulating the JAK-STAT signalling pathway.

INTRODUCTION: Searching for new therapeutic possibilities constitutes a challenge for modern medicine and an answer to better understanding of molecular mechanisms of pro-inflammatory diseases. The JAK-STAT pathway plays an important role in the inflammatory processes, which is supported by the fact that its inhibitors are used to treat, for instance, psoriasis and rheumatoid arthritis. AIM: To determine whether the epigenetic mechanisms - methylation of gene promotion regions and miRNAs may serve as a new therapeutic strategy for JAK-STAT pathway inhibition. MATERIAL AND METHODS: Basing on MethPrimer (plus CpG Island Prediction) program and microrna.org database of the said mechanism in the regulation of the JAK-STAT signalling pathway, the gene expression was performed, indicating or excluding the possibility of their use as new potential therapeutic strategies. RESULTS: A different number of CpG islands (CGI) for each gene (JAK1-4 CGI; JAK2-2 CGI; JAK3-5 CGI, TYK2-6 CGI; STAT1-2 CGI; STAT2-1 CGI; STAT3-3 CGI; STAT5A-4 CGI; STAT5B-3 CGI) might be a new therapeutic goal. What is more, our results show that genes associated with JAK-STAT signalling pathways can be regulated by miRNAs (JAK1-42 miRNAs; JAK2-47 miRNAs; JAK3-15 miRNAs, TYK2-4 miRNAs; STAT1-17 miRNAs; STAT2-30 miRNAs, STAT3-36 miRNAs, STAT4-15 miRNAs; STAT5A-10 miRNAs; STAT5B-23 miRNAs). CONCLUSIONS: The epigenetic mechanisms of the regulation of the JAK-STAT signalling pathway gene expression constitute a promising new therapeutic strategy for treatment of those diseases, during which disorders are observed in gene expression models of the analysed signalling pathway.

The Influence of Salinomycin on the Expression Profile of mRNAs Encoding Selected Caspases and MiRNAs Regulating their Expression in Endometrial Cancer Cell Line.

BACKGROUND: Apoptosis could take place in the pathway dependent on death receptors or pathways dependent on mitochondria. In both, a key role is played by enzymes with protease activity, known as caspases. AIM: The aim of this study was to assess the variances in the expression pattern of caspase-dependent signaling pathways in the endometrial cancer cell line when treated with salinomycin. Additionally, the changes in the level of miRNA that potentially regulate these mRNAs were evaluated. MATERIALS AND METHODS: Endometrial cancer cells were treated with 1 µM of salinomycin for 12, 24 and 48 hours. Untreated cells made up the control culture. The molecular analysis consisted of screening mRNA and miRNA microarray expression profiles of caspases, and the evaluation of the expression of caspases 3,8 and 9 by RTqPCR, also on the protein level. RESULTS AND DISCUSSION: It was observed that 5 of the 14 differentiating mRNAs were commonly found for all incubation times of the cells and they corresponded with CASP3, CASP8, and CASP9 genes. The highest impact probability was determined between CASP3(up-regulated) and hsa- miR- 30d (FC -2.01), CASP8 (down-regulated) and hsa-miR-21 (FC +1.39) and between CASP9 (upregulated) and hsa-miR-1271 (FC +1.71). CONCLUSION: Salinomycin induces the apoptosis of endometrial cancer cells. The largest increase in activity was noted for caspases 3 and 9, while the expression of caspase 8 was decreased. Salinomycin causes a regulatory effect on the transcriptomes of mRNA and miRNA in in vitro endometrial cancer cells.

Cisplatin Changes Expression of SEMA3B in Endometrial Cancer.

BACKGROUND: Semaphorin 3B (SEMA3B) is characterized as a strong suppressing factor of the proliferation of cancerous cells and also by its anti-angiogenic effect. However, the knowledge on the changes in the expression profile of SEMA3B under the influence of cisplatin in endometrial cancer remains fragmented. The aim of this work was to note the changes in expression of SEMA3B when under the influence of cisplatin in the endometrial cancer cell line. METHODS: Ishikawa cell line cells were exposed to three different concentrations of cisplatin: 2.5µM; 5µM; 10µM for 12, 24 and 48 hours and were compared to cells untreated by the drug. Changes in the expression profile of SEMA3B were determined based upon RtqPCR (mRNA) alongside the ELISA assay (protein). The Statistica 13.0 PL program was used for statistical analysis (p<0.05). RESULTS: Changes on the transcriptome level seem to be more dynamic than on the proteome level. Regardless of the concentration given or the exposition period, the expression of semaphorin 3B was, in fact, higher in cells exposed to cisplatin. Statistically substantial differences (p<0.05) in the expression of SEMA3B mRNA and protein were seen for all incubation periods at the given cisplatin level when compared to the control. CONCLUSION: Cisplatin causes a growth in the expression of SEMA3B in an endometrial cancer cell culture, this results in the restoration in the state of cell homeostasis and shows the effectiveness of pharmacotherapy, including a low risk of drug resistance.

Effect of Salinomycin on Expression Pattern of Genes Associated with Apoptosis in Endometrial Cancer Cell Line.

BACKGROUND: Salinomycin is part of a group of ionophore antibiotics characterized by an activity towards tumor cells. To this day, the mechanism through which salinomycin induces their apoptosis is not fully known yet. The goal of this study was to assess the expression pattern of genes and the proteins coded by them connected with the process of programmed cell death in an endometrial cancer cell Ishikawa culture exposed to salinomycin and compared to the control. MATERIALS AND METHODS: Analysis of the effect of salinomycin on Ishikawa endometrial cancer cells (ECACC 99040201) included a cytotoxicity MTT test (with a concentration range of 0.1-100 µM), assessment of the induction of apoptosis and necrosis by salinomycin at a concentration of 1 µM as well the assessment of the expression of the genes chosen in the microarray experiment (microarray HG-U 133A_2) and the proteins coded by them connected with apoptosis (RTqPCR, ELISA assay). The statistical significance level for all analyses carried out as part of this study was p<0.05. RESULTS: It was observed that salinomycin causes the death of about 50% of cells treated by it (50.74±0.80% of all cells) at a concentration of 1µM. The decrease in the number of living cells was determined directly after treatment of the cells with the drug (time 0). The average percent of late apoptotic cells was 1.65±0.24% and 0.57±0.01% for necrotic cells throughout the entire observation period. DISCUSSION: Microarray analysis indicated the following number of mRNA differentiating the culture depending on the time of incubation with the drug: H_12 vs C = 114 mRNA, H_8 vs C = 84 mRNA, H_48 vs. C = 27 mRNA, whereas 5 mRNAs were expressed differently at all times. During the whole incubation period of the cells with the drug, the following dependence of the expression profile of the analyzed transcripts was observed: Bax>p53>FASL>BIRC5>BCL2L. CONCLUSION: The analysis carried out indicated that salinomycin, at a concentration of 1 µM, stopped the proliferation of 50% of endometrial cancer cells, mainly by inducing the apoptotic process of the cells. The molecular exponent of the induction of programmed cell death was an observed increase in the transcriptional activity of pro-apoptotic genes: Bax;p53;FASL and a decrease in the expression of anti-apoptotic genes: BCL2L2; BIRC5.

Evaluation of Changes in the Expression Profile of mRNA and Proteinencoding Adiponectin in Ishikawa Cell Line under the Influence of Cisplatin - Preliminary Report.

BACKGROUND: A reduced concentration of adiponectin is considered as an independent factor of the risk of inducing endometrial cancer. Cisplatin is a drug used in the therapy of this type of neoplasm. However, knowledge of the effects of cisplatin on the adiponectin level is still limited. OBJECTIVE: The purpose of this study was to assess the impact of cisplatin depending on the concentration and time of exposition of the cells to the drug on the adiponectin level in the endometrial cancer cell line. METHODS: Cells of endometrial cancer cell line Ishikawa were exposed for 12,24 and 48 hour periods to cisplatin with the following concentrations: 2.5µM, 5µM, 10µM. The changes in the expression profile of adiponectin were compared to the RtqPCR reaction and ELISA test. The STATISTICA 13.0 PL program was used for statistical analysis (p<0.05). RESULTS: In the culture without the drug, the concentration of adiponectin was statistically lower than in the cell culture incubated with the drug. Changes on the mRNA level seem to be more specific than on the protein level, although in both cases, the same trend in the expression changes was noted. DISCUSSION: The longer the time of exposition of the cells to the drug, the expression of mRNA, and the adiponectin protein increased. Changes in the expression profile were characterized statistically (p<0.05). CONCLUSION: Cisplatin, in a noticeable way, changes the expression profile of adiponectin. Molecular analysis indicated that in the case of endometrial cancer therapy should be implemented with a concentration of no less than 5 µM.

Differences in the Expression Pattern of mRNA Protein SEMA3F in Endometrial Cancer in vitro under Cisplatin Treatment.

BACKGROUND: Semaphorin 3F (SEMA3F) plays a substantial role in carcinogenesis, because of its role in inducing angiogenesis, and creating a microenvironment for the developing tumor. OBJECTIVE: The purpose of this work was to assess the impact of cisplatin, depending on the concentration and exposure time on the expression pattern of SEMA3F in an endometrial cancer cell line. MATERIALS AND METHODS: Cultures of the Ishikawa endometrial cancer cells were incubated with cisplatin with the following concentrations: 2.5µM; 5µM; and 10µM and for the following periods of time: 12; 24; and 48 hours. Cells not incubated with the drug constituted the control in the experiment. To determine the effect of cisplatin on the expression of SEMA3F, the real-time quantitative reverse transcription reaction (RtqPCR; mRNA) was used, as well as the ELISA assay (protein). The statistical analysis was done with the admission of p<0.05. RESULTS: The silencing of SEMA3F expression on the transcriptome and proteome levels in a culture unexposed to the effects of cisplatin in comparison to endometrial cancer cells under the influence of cisplatin (p<0.05) were noted. Along with an increase in the concentration of the drug used, the number of copies of the gene transcript, during the shortest incubation period had a gradual increase. Only for the highest concentration of the drug, substantial statistical differences in the expression of the SEMA3F protein between 24 and 48 hour incubation periods (p<0.05) were determined. CONCLUSION: Using cisplatin in an endometrial cancer cell culture results in an increased expression of SEMA3F, which advantageously affects the normalization of the neoplastic angiogenic process and lowers the proliferation of the cells making up the mass of the tumor.

Effect of adalimumab on the expression profile of mRNA, and protein associated with JAK/STAT signaling pathway in fibroblast exposed to lipopolysaccharide.

The aim of this study was to assess the effect of adalimumab on the expression level of mRNA and protein TNF-α, IFN-γ, IL-17, IL12A, IL12B, and IL23A in the culture of normal human fibroblasts, in which the LPS inflammation process was induced. The NHDF culture was exposed to the effect of LPS in the concentrations of 1, 2, and 10 µg/mL for 2, 8, and 24 hour periods, and then adalimumab was added at the concentration of 8 µg/mL, it was then incubated for 2, 8, and 24 hour. Cells unexposed to LPS and adalimumab constituted the control. The microarray expression techniques, RTqPCR, and ELISA assay were used. Irrespectively of the concentration of LPS used and the incubation time of it with cells overexpression of the analyzed genes is present, with increasing factor concentration used to induce inflammation and incubation time with it, expression of the assessed genes was greater. In turn, adding the anti-TNF drug to the culture caused the silencing of the expression of the mRNAs and proteins. It was confirmed that LPS and adalimumab above all affect the expression of genes and proteins dependent on the interaction of IL-12 with receptors, which are TNF-α and IFN-γ, and to a lesser extent also modulate IL-17.

Is ursodeoxycholic acid therapy due to pregnant intrahepatic cholestasis change the adiponectin and fibroblast growth factor-21 levels?-Pilot study.

The main aim of the study was to assess changes in adiponectin and fibroblast growth factor 21 (FGF21) levels in pregnant women with intrahepatic cholestasis during ursodeoxycholic acid (UDCA) therapy. The study included 40 pregnant women with intrahepatic cholestasis (ICP) treated with UDCA. In the pregnant ICP group, material for further analysis was collected three times: before the first dose of drug T1, 4 weeks after the first dose of drug T2, 8 weeks after the first dose of drug T3, and 1 day after delivery T4 (P < .05). Regarding changes in the adiponectin concentration profile, three statistical significance (P < .05) was found: before the first dose and 8 weeks of treatment and 1 day after delivery, as well as between 4 and 8 weeks of UDCA acid therapy. In the fourth and eighth weeks of treatment, adiponectin levels reached a higher concentration than before the first dose of UDCA, but a decrease was observed 1 day after delivery. It has been confirmed that UDCA therapy has an impact on the dynamics of changes in adiponectin and FGF21 levels as well as indicators characterizing liver function.

Effect of ursodeoxycholic acid therapy due to pregnant intrahepatic cholestasis on chemerin and irisin levels.

The purpose of the work was to assess changes in chemerin and irisin levels in women with diagnosed intrahepatic cholestasis of pregnant women treated with ursodeoxycholic acid. The study group consisted of 50 patients with diagnosed and confirmed intrahepatic cholestasis of pregnant women at 24-25 weeks of pregnancy treatment by ursodeoxycholic acid (UDCA). The study also included a group of 40 pregnant women, without concomitant intrahepatic cholestasis of pregnancy (ICP). In the pregnant ICP group, whole blood was collected 4 times: before the first dose of drug, 4 and 8 weeks after the first dose, and day after delivery. It was observed that statistically significant differences in the concentration of irisine occur between the time before starting treatment and the 8-week therapy and 1 day after delivery. The Pearson correlation analysis (r's) showed two statistically significant relationships (p < .05). The first of these can be found between the concentration of irisine and chemerin in the group of nonpregnant women and the second in the group of patients with intrahepatic pregnant cholestasis before the first dose of UDCA. A significant relationship between irisin and chemerin concentrations was confirmed in the group of pregnant ICP patients during UDCA acid therapy and among healthy pregnant women.

Variances in the Level of COX-2 and iNOS in Different Grades of Endometrial Cancer.

BACKGROUND: Many experimental studies have demonstrated the importance of COX-2 in the tumor angiogenesis. Inducible iNOS is responsible for a high and stable level of nitric oxide and is expressed in response to pro-inflammatory factors. OBJECTIVE: The aim of this study was to evaluate the expression of COX-2 and iNOS at the protein level and to assess their potential prognostic significance in patients with endometrial cancer. METHODS: The study group consisted of 45 women with endometrial cancer divided according to the degree of histological differentiation i.e. G1, 17; G2, 15; G3, 13. The control group consisted of 15 women without neoplastic changes. The expression of studied proteins was determined immunohistochemically with specific polyclonal antibodies. RESULTS: Analysis of the COX-2 expression showed that the optical density of the reaction product in G1 reached 186% in the control group, while the values in G2 and G3 reached 243% and 293%, respectively. In the case of iNOS, the optical density of the reaction product reached the following percentages in the control group: 147% in G1, 243% in G2, and 241% in G3. CONCLUSION: Our findings suggest that changes in the expression of COX-2 and iNOS may be potentially useful in predicting the progression of endometrial cancer and treatment effectiveness.