Spontaneous regression of malignant melanoma - is it based on the interplay between host immune system and melanoma antigens?

Malignant melanoma (MM) is the most aggressive and uneasily treatable form of skin cancer. Up to 90% of deaths because of skin tumours are estimated to be caused by this malignancy. Spontaneous regression is described as a partial or complete disappearance of cancer. It can be defined if the clinical and histological diagnosis of malignancy is verified and any therapeutic intervention potentially inducing mechanisms leading to regression has not been applied. Regression occurs more frequently in melanoma than in other types of tumours; it is reported to be six times higher than in other malignancies. Up to 50% of primary MM is reported to undergo spontaneous regression. However, spontaneous regression of the metastatic form of tumour is a rare phenomenon observed in only 0.23% of cases. The most frequently mentioned factors leading to spontaneous regression of MM are operative trauma, infection, vaccination (BCG and rabies vaccines) and immunological factors. Other well-documented circumstances associated with regression of metastatic MM include blood transfusion and various endocrine factors.

Spontaneous regression of tumour and the role of microbial infection--possibilities for cancer treatment.

This review deals with the role of microorganisms in spontaneous regression of a tumour. Spontaneous cancer regression is a phenomenon that has been described for many centuries. One of the most well known methods of inducing spontaneous regression of cancer is the application of Coley's toxin (heat-killed Streptococcus pyogenes and Serratia marcescens), which has been used for the successful treatment of sarcomas, carcinomas, lymphomas, myelomas and melanomas. In clinical practice, the use of Bacillus Calmette-Guérin vaccine for the treatment of superficial urinary bladder cancer is the most common instance of the application of microorganisms for the treatment of cancer. This review provides further information on other tested bacteria--Clostridium spp., Bifidobacterium spp., Lactobacillus spp. and Salmonella spp.--in this field of study. Among new age methods, bactofection, alternative gene therapy, combination bacteriolytic therapy and bacteria-directed enzyme prodrug therapy are some of the potential cancer treatment modalities that use microorganisms. We have also provided information about the interconnection among microorganisms, immune system response, and the possible mechanisms involved in the spontaneous regression of tumours.

Two-step separation of nostotrebin 6 from cultivated soil cyanobacterium (Nostoc sp.) by high performance countercurrent chromatography.

High performance countercurrent chromatography (HPCCC) was successfully applied for the separation of nostotrebin 6 from cultivated soil cyanobacteria in a two-step operation. A two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (4:5:4:5, v/v/v/v) was employed for the HPCCC separation. In the first-step operation, its neutral upper phase was used as stationary phase and its basic lower phase (1% NH3 in lower phase) was employed as mobile phase at a flow rate of 1 mL/min. In the second operation step, its neutral upper phase was used as stationary phase, whereas both its neutral lower phase and basic lower phase were employed as mobile phase with a linear gradient elution at a flow rate of 0.8 mL/min. The revolution speed and temperature of the separation column were 1,000 rpm and 30 °C, respectively. Using HPCCC followed by clean-up on Sephadex LH-20 gel, 4 mg of nostotrebin 6 with a purity of 99% as determined by HPLC/DAD-ESI-HRMS was obtained from 100 mg of crude extract. The chemical identity of the isolated compound was confirmed by comparing its spectroscopic data (UV, ESI-HRMS, ESI-HRMS2) with those of an authentic standard and data available in the literature.

Treatment of thyroid disorder supported by 5-phosphodiesterase inhibitors improved erectile dysfunction in patients with hypo- and hyperthyroidism.

INTRODUCTION: The relationship between a poorly functioning thyroid gland and erectile dysfunction (hereinafter, "ED") has been demonstrated in many studies. If a man has thyroid problems, it can negatively affect his ability to achieve or maintain an erection. The thyroid gland produces hormones that affect metabolism and overall body function, including blood circulation. If the thyroid gland is not functioning properly, it can lead to blood circulation disorders, which can affect erectile function. There are also symptoms of thyroid disorders that can contribute to ED. Some of these symptoms include fatigue, depression, and anxiety, which can negatively affect a man's psychological side and affect his sexual performance. OBJECTIVES AND METHODS: In our article, we present a series of patients with ED who were treated in our department and found to have some form of thyroid dysfunction. RESULTS: After treatment for thyroid dysfunction and treatment with 5-phosphodiesterase inhibitors were implemented, erectile function improved in all patients ((Wilcoxon Signed Rank Test; Z = -4.55; p (2-tailed) < 0.001; n = 27) to the level of mild or no ED. Improvement occurred in men with hyper- and hypothyroidism. After one year of treatment, there was no difference between the two groups (t-test; t = 0.75; df = 0.25; p < 0.46). CONCLUSION: This study shows that screening for thyroid dysfunction should be performed in all men with ED and that treating thyroid dysfunction may be an effective way to improve erectile function in men with these health problems.

Real-time PCR method for the detection of the gene encoding surface lipoprotein LipL32 of pathogenic Leptospira: use in the laboratory diagnosis of the acute form of leptospirosis.

BACKGROUND: The aims of this work were to replace the obsolete PCR method for the laboratory diagnosis of the acute form of leptospirosis using the G1, G2 and B64 I, B64 II primers, and to improve the PCR detection time. METHODS: We introduced a real-time PCR method for the detection of the gene encoding the surface lipoprotein LipL32 of pathogenic Leptospira into our laboratory diagnosis of the acute form of leptospirosis. The positive and negative analytical specificities of the real-time PCR method were both equal to 100%; the detection limit was determined to be 1-5 genome copies/1 ml of liquid biological material. The method was further validated on 230 laboratory strains of leptospires. RESULTS: All laboratory strains of pathogenic Leptospira were evaluated as LipL32-positive and all non-pathogenic strains as LipL32-negative. In addition, 455 biological materials (253 plasma, 121 urine, 72 cerebrospinal fluid (CSF), 7 bronchoalveolar lavage, and 2 sputum) from 295 patients with suspected leptospirosis were examined. From this set of patients, 9 were evaluated to be LipL32-positive, from 15 positive biological materials (10 urine, 4 blood plasma, and 1 CSF). CONCLUSIONS: This real-time PCR method for the detection of the gene encoding the surface lipoprotein LipL32 is a reliable, sensitive, and rapid method for the detection of the acute form of leptospirosis.

Determination of substitution sites in monosubstituted five-membered aromatic heterocycles.

Similar magnitudes of proton-proton couplings across three, four, and five bonds and proton-carbon couplings across two and three bonds combined with difficult to predict substituent effects make the results of an indiscriminate use of routine (COSY, HSQC, HMBC, etc.) techniques for substitution site determination in C-monosubstituted five-membered heteroaromatics suspect. As demonstrated on two examples of natural products, the use of 1,1-ADEQUATE leads to unambiguous substitution site determination lending thus further support to suggested inclusion of 1,1-ADEQUATE data into computer-assisted structure elucidation (CASE) protocols.

[Comparison of results of two serological methods for diagnosing leptospirosis - microagglutination test and ELISA]. / Porovnání výsledku dvou metod sérologických diagnostiky leptospirózy - mikroaglutinacniho testu a metody ELISA.

BACKGROUND: The aim of this study was to evaluate diagnostic sensitivity and specificity of the SERION ELISA classic IgM and SERION ELISA classic IgG kits and to confirm the results by the microagglutination test (MAT). MATERIAL AND METHODS: A total of 45 blood serum samples from 45 patients, 30 from males and 15 from females (mean age 44.24 ± 15.56 range 19-82 year), were included in our study. Blood serum samples were examined using the ELISA and MAT methods and diagnostic sensitivity and specificity of both methods were calculated. RESULTS: The MAT was shown to have 100 % diagnostic sensitivity and specificity. The ELISA kits for detecting IgM and IgG antibodies against pathogenic leptospires had diagnostic sensitivity of 100 % and diagnostic specificity of 88.6 % and 54.3 %, respectively. CONCLUSION: The above results suggest that the MAT with diagnostic sensitivity and specificity of 100 % remains the gold standard for detection of specific antibodies against pathogenic leptospires. The diagnostic sensitivity of both ELISA kits is high but due to their low diagnostic specificity, especially in the case of IgG antibodies, the kits are inappropriate for use in routine clinical practice.

[Incidence of leptospirosis in Pardubice and Hradec Králové region and a part of Vysocina region in the years 2002-2009]. / Výskyt leptospirózy v Kralovehradeckem a Pardubické m kraji a v cásti kraje Vysocina v letech 2002-2009.

BACKGROUND: The aim of this study was to evaluate the incidence of leptospirosis in Pardubice and Hradec Králové regions and a part of Vysocina region in 2002-2009. A group of patients was statistically analyzed and the incidence rates of leptospirosis in individual months and years were calculated. MATERIAL AND METHODS: A group of 4,813 patients with suspected leptospirosis from Pardubice and Hradec Králové regions and a part of Vysocina region were examinated in our centre in 2002-2009. A total of 5,366 biological samples were studied. From 4,625 blood samples, 3,958 were examinated with the serological microagglutination-lysis method and the remaining 667 with the PCR method. RESULTS: From 4,813 patients suspected to have leptospirosis, 89 were found to be leptospirosis-positive. Leptospirosis was diagnosed in 63 males (70.7 %) a 26 females (29.3 %); the mean age were 39.52 ± 17.67 years in males and 46.85 ± 18.34 years in females; the mean age men + women was 41.66 ± 18.13 years (range 3-78 years). Most positive male patients (n = 28) were in 26-45 age group; the findings were similar in females (n = 10). The highest number of leptospirosis cases was noted in 2005. Most frequently, infection caused by pathogenic leptospires was diagnosed in November. CONCLUSION: The incidence rates of leptospirosis in Pardubice and Hradec Králové regions and a part of Vysocina region in 2002-2009 (with the exception of 2008) exceeded the overall reported levels 0.4/100,000 population.

Congenital Hyperinsulinism Caused by Novel Homozygous KATP Channel Gene Variants May Be Linked to Unexplained Neonatal Deaths among Kurdish Consanguineous Families.

INTRODUCTION: Neonatal hypoglycemia due to congenital hyperinsulinism (CHI) is a potentially life-threatening condition. Biallelic pathogenic variants in KATP channel subunit genes (ABCC8, KCNJ11), causing severe forms of CHI, are more prevalent in regions with a significant rate of consanguinity and may lead to unexplained neonatal deaths. We hypothesized that KATP channel gene variants are the cause of CHI in three unrelated children from consanguineous Kurdish families with histories of four unexplained neonatal deaths with convulsions. CASES: (1) A girl presented on the 6th day of life with recurrent hypoglycemic convulsions (blood glucose 2.05 mmol/L, insulin 58 mIU/L, C-peptide 2,242 pmol/L). (2) A girl with severe developmental delay was diagnosed with CHI at 3 years of age (blood glucose 2.78 mmol/L, insulin 8.1 mIU/L, C-peptide 761 pmol/L) despite a history of recurrent hypoglycemia since neonatal age. (3) A girl presented at 3 weeks of age with convulsions and unconsciousness (blood glucose 2.5 mmol/L, insulin 14.6 mIU/L, C-peptide 523 pmol/L). Coding regions of the ABCC8 and KCNJ11 genes were tested by Sanger sequencing. Potential variants were evaluated using the American College of Medical Genetics standards. Three novel causative homozygous variants were found - p.Trp514Ter in the ABCC8 gene (Pt2), and p.Met1Val (Pt1) and p.Tyr26Ter (Pt3) in the KCNJ11 gene. CONCLUSION: CHI caused by KATP channel variants was elucidated in three children, providing a highly probable retrospective diagnosis for their deceased siblings. Future lives can be saved by timely diagnosis of CHI when encountering a neonate with unexplained seizures or other signs of recurrent and/or persistent hypoglycemia.

NPR2 Variants Are Frequent among Children with Familiar Short Stature and Respond Well to Growth Hormone Therapy.

CONTEXT: The C-type natriuretic peptide receptor encoded by the NPR2 gene is a paracrine regulator of the growth plate; heterozygous NPR2 variants cause short stature with possible presence of different signs of bone dysplasia. To date, the effect of growth hormone (GH) treatment has been described in a few individuals with NPR2 gene variants with inconsistent results. OBJECTIVES: To identify NPR2 gene variants among children with familial short stature (FSS) and to describe their phenotype, including GH treatment response. DESIGN, SETTINGS AND PATIENTS: Out of 747 patients with short stature treated with GH in a single center, 87 with FSS met the inclusion criteria (pretreatment height ≤ -2 standard deviation in both the patient and the shorter parent, unknown genetic etiology). Next-generation sequencing methods were performed to search for NPR2 gene variants. The results were evaluated using the American College of Medical Genetics and Genomics guidelines. The GH treatment response (growth velocity improvement and height standard deviation score development over the first 5 years of treatment) was evaluated. RESULTS: In 5/87 children (5.7%), a (likely) pathogenic variant in the NPR2 gene was identified (p.Ile558Thr [in 2], p.Arg205*, p.Arg557His, p.Ser603Thr). Two children had disproportionate short-limbed short stature, 1 a dysplastic 5th finger phalanx. The growth velocity in the first year of GH treatment accelerated by 3.6 to 4.2 cm/year; the height improved by 1.2 to 1.8 SD over 5 years of treatment. CONCLUSIONS: NPR2 gene variants cause FSS in a significant proportion of children. Their GH treatment response is promising. Studies including final height data are necessary to assess the long-term efficacy of this therapy.

Melanoma-related changes in skin microbiome.

Melanoma is the least common form of skin tumor, but it is potentially the most dangerous and responsible for the majority of skin cancer deaths. We suggest that the skin microbiome might be changed during the progression of melanoma. The aim of this study is to compare the composition of the skin microbiota between different locations (skin and melanoma) of a MeLiM (Melanoma-bearing Libechov Minipig) pig model (exophytic melanoma). Ninety samples were used for PCR-DGGE analysis with primers specifically targeting the V3 region of the 16S rRNA gene. The profiles were used for cluster analysis by UPGMA and principal coordinate analysis PCoA and also to calculate the diversity index (Simpson index of diversity). By comparing the obtained results, we found that both bacterial composition and diversity were significantly different between the skin and melanoma microbiomes. The abundances of Fusobacterium and Trueperella genera were significantly increased in melanoma samples, suggesting a strong relationship between melanoma development and skin microbiome changes.

Antiinflammatory and neurological activity of pyrithione and related sulfur-containing pyridine N-oxides from Persian shallot (Allium stipitatum).

ETHNOPHARMACOLOGICAL RELEVANCE: Persian shallot (Allium stipitatum) is a bulbous plant native to Turkey, Iran and Central Asia. It is frequently used in folk medicine for the treatment of a variety of disorders, including inflammation and stress. Antiinflammatory and neurological activities of pyrithione and four related sulfur-containing pyridine N-oxides which are prominent constituents of Allium stipitatum were tested. METHODS: The antiinflammatory activity was tested by the ability of the compounds to inhibit cyclooxygenase (COX-1 and COX-2), whereas the neurological activities were evaluated by assessing the compounds ability to inhibit monoamine oxidase-A (MAO-A) and acetylcholinesterase (AChE). The compounds׳ affinity for the serotonin transport protein (SERT) and the GABAA-benzodiazepine receptor were also investigated. RESULTS: 2-[(Methylthio)methyldithio]pyridine N-oxide showed very high antiinflammatory effects which are comparable with those of common pharmaceuticals (IC50 of 7.8 and 15.4 µM for COX-1 and COX-2, respectively). On the other hand, neurological activities of the compounds were rather modest. Some compounds moderately inhibited AChE (IC50 of 104-1041 µM) and MAO-A (IC50 of 98-241 µM) and exhibited an affinity for the SERT and GABAA-benzodiazepine receptor. CONCLUSIONS: Our findings may help to rationalize the wide use of Persian shallot for the treatment of inflammatory disorders.

Our experience using real-time PCR for the detection of the gene that encodes the superficial lipoprotein LipL32 of the pathogenic leptospires to confirm the acute form of human leptospirosis.

AIMS: To examine biological materials (blood, urine, cerebrospinal fluid) of patients with suspected leptospirosis using real-time PCR for detecting the gene that codes the superficial LipL32 lipoprotein, and to evaluate the contribution of the real-time PCR method for the laboratory diagnosis of the acute form of leptospirosis. MATERIAL AND METHODS: During the monitored period of April 2010 - December 2011, a total of 340 biological materials samples were examined (177x blood plasma, 88x urine, 68x, cerebrospinal fluid, 6x bronchoalveolar lavage and 1x sputum) from 216 patients with suspected leptospirosis using real-time PCR LipL32 gene detection. RESULTS: From the mentioned 216 patients suspected of leptospirosis, 8 patients were evaluated as being PCR LipL32 positive, from which 14 positive biological materials originated (9 x urine, 4x blood and 1x liquor). CONCLUSION: As demonstrated in the study, the real-time PCR method for detecting the gene for the superficial lipoprotein LipL32 is an appropriate, quick and reliable method for the diagnosis of the acute form of leptospirosis.

Allium discoloration: the precursor and formation of the red pigment in giant onion (Allium giganteum Regel) and some other subgenus Melanocrommyum species.

The precursor of the orange-red pigment formed upon wounding the bulbs of Allium giganteum (Allium subg. Melanocrommyum) was isolated and shown to be S-(2-pyrrolyl)cysteine S-oxide. In addition, two other pyrrolylsulfinyl derivatives were found in an extract from the bulbs, namely, 3-(2-pyrrolylsulfinyl)lactic acid and S-(3-pyrrolyl)cysteine S-oxide. Contrary to a previous report, the latter compound was shown not to serve as the precursor of the pigment, being in fact only an artifact formed during isolation. The formation of pyrrolyl-containing compounds following disruption of A. giganteum bulbs was studied by a combination of LC-MS, LC-NMR and DART-MS. It was found that S-(2-pyrrolyl)cysteine S-oxide is cleaved by a C-S lyase (alliinase) to yield 2-pyrrolesulfenic acid. Two molecules of the latter compound give rise to highly reactive S-(2-pyrrolyl) 2-pyrrolethiosulfinate which in turn converts into red 2,2'-epidithio-3,3'-dipyrrole (dipyrrolo[2,3-d:2',3'-e]-1,2-dithiin). Several other pyrrolyl-containing compounds were detected in A. giganteum for the first time, including S-methyl 2-pyrrolethiosulfinate, S-(2-pyrrolyl) methanethiosulfinate, di(2-pyrrolyl) disulfide, and S-(2-pyrrolyl) 2-pyrrolethiosulfonate. It can be concluded that the formation of the orange-red pigment in Allium subg. Melanocrommyum species, despite sharing several analogous features, is of a different nature than the pink discoloration of onion (A. cepa).