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One of the most difficult in diagnostic and treatment options for endogenous Cushing is the ectopic ACTH syndrome, which causes the development of tumors of different histogenesis localization producing adrenocorticotropic hormone (ACTH), and much less - corticotropin hormone (CRH). ACTH-secreting tumors varied in location, morphological structure and the degree of malignancy. Most of these tumors are characterized by an aggressive course with a propensity to metastasize and relapse. The article presents data of the prevalence, pathogenesis of ectopic ACTH tumors localized in the thymus, analyzis of clinical, morphological features, the methods of diagnosis and treatment. Based on the current literature, the world and our own experience on the diagnosis and treatment of patients with ectopic ACTH syndrome with localization of hormone production in the thymus, we want to highlight the current state of the problem in order to create the most efficient algorithm for diagnostic search and treatment of this difficult group of patients.

At the present time topical diagnosis of ACTH-producing neuroendocrine tumors of different localization is the most challenging problem for endocrinologists around the world. Despite the significant improvement of existing diagnostic techniques, in 9-19% localization of the source of ectopic secretion ACTH re-mains undetected throughout life. Thus even regular using of routine laboratory tests and visual methods of diag- nosis do not allow to reveal the primary tumor. This article describes a case report, that is particular by a long period of ectopic ACTH syndrome detection. However, this demonstrates the importance of inten-sive and prolonged diagnosis of primary tumor, that can significantly reduce the morbidity and mortality of these patients and improve their prognosis.

Ectopic hormonesecreting of cecum are rare. We report the case of a 52yearold woman with a typical clin ical picture of Cushing disease. Diagnostic tests confirmed ACTH dependent Cushing’s syndrome, which is caused by neuroendocrine tumor of the cecum. Due to the rarity of the disease, we believe it appropriate to share our experience of this group of patients.

Ectopic hormone-secreting pheochromocytomas are rare. Only case reports exist in the literature. Despite the large number of guides on diagnosis and treatment of pheochromocytoma, and Cushing syndrome, the extreme rarity of ectopic ACTH-syndrome caused by pheochromocytoma, and complexity of clinical cause numerous diagnostic errors leading to treatment failure. Therefore, we belive it appropriate to share our experience of this group of patients.

A 31-year-old male with a history of obesity, type 2 diabetes mellitus (T2DM) and hypercholesterolemia came to our clinic for medical weight-loss management. According to his initial anthropometric measurements (weight - 193 kg, height - 181 cm, and body mass index (BMI) - 58 kg/m2) patient had morbid obesity. In order to educate our patient about good nutrition, physical activity and the need of sun exposure we used an online system based on the video lessons, full of humor, pictures, and cartoons. Even one month of watching the short educational movies was sufficient for the patient to form the habits of healthy eating, including restriction of fat and digestible carbohydrates and increase in daily consumption of low-fat dairy products, slow carbohydrates, protein and fiber. Endocrinologist was regularly available for any questions and inquiries that the patient might have had. In addition to the dietary recommendations subcutaneous liraglutide 2.4 mg daily was initiated. After 8 months of treatment the patient has lost 58 kg and reached the body weight of 135 kg; moreover, he had motivation to continue losing weight.

Large insert genomic DNA libraries are useful resources for genomic studies. Although the genome of Xenopus tropicalis stands as the amphibian reference genome because it benefitted from large-scale sequencing studies, physical mapping resources such as BAC libraries are lagging behind. Here we present the construction and characterization of a BAC library that covers the whole X. tropicalis genome. We prepared this BAC library from the genomic DNA of X. tropicalis females of the Adiopodoume strain. We characterized BAC clones by screening for specific loci, by chromosomal localization using FISH and by systematic BAC end sequencing. The median insert size is about 110kbp and the library coverage is around six genome equivalents. We obtained a total of 163,787 BAC end sequences with mate pairs for 77,711 BAC clones. We mapped all BAC end sequences to the reference X. tropicalis genome assembly to enable the identification of BAC clones covering specific loci. Overall, this BAC library resource complements the knowledge of the X. tropicalis genome and should further promote its use as a reference genome for developmental biology studies and amphibian comparative genomics., (Copyright © 2017 Elsevier Inc. All rights reserved.)

A new sensitive non-invasive gold nanoparticle-based sensor that enables to detect thiols in the human skin has been developed. The detection procedure implied the assessment of the color change of a paper sensor resulting from aggregation of gold nanoparticles caused by thiols. The ratio of the intensity of the photo image blue channel vs the red one (in units of RGB coloration) served as analytical response. The main thiol in the skin is glutathione, therefore, it was used as model biothiol and spiking substance. The range of linearity for glutathione was 8-75µM, the detection limit was 6.9µM. RSD≤7% is for inter-day determination of 10μM glutathione and RSD≤12% is the intra-day value. The recovery of 5µM and 10µM of glutathione was evaluated by applying solution, containing thiol-spikes, on skin. The results varied in the range 77-138%. A hundred-fold excess of serine, alanine, histidine, threonine, creatinine, urea, and ammonia; a ten-fold excess of glycine, proline, leucine, isoleucine, phenylalanine, asparagine; and a five-fold excess of valine, tryptophan, tyrosine, and uric acid, which can be extracted from the skin and is contained in the test matrix, have no significant effect on 10µM glutathione signal. Thiols level in the skin of volunteers (21-65 years old, men and women) detected with the use of a proposed non-invasive sensor was 11.6-47.5µM., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Aims: Salmonella is a worldwide foodborne pathogen causing acute enteric infections in humans. In the recent years, the use of bacteriophages has been suggested as a possible tool to combat this zoonotic pathogen in poultry farms. This work aims to isolate and perform comparative studies of a group of phages active against a collection of specific Salmonella Enteritidis strains from Portugal and England. Also, suitable phage candidates for therapy of poultry will be selected., Methods and Results: The Salm. Enteritidis strains studied were shown to have a significantly high occurrence of defective (cryptic) prophages; however, no live phages were found in the strains. Bacteriophages isolated from different environments lysed all except one of the tested Salm. Enteritidis strains. The bacteriophages studied were divided into different groups according to their genetic homology, RFLP profiles and phenotypic features, and most of them showed no DNA homology with the bacterial hosts. The bacteriophage lytic efficacy proved to be highly dependent on the propagation host strain., Conclusions: Despite the evidences shown in this work that the Salm. Enteritidis strains used did not produce viable phages, we have confirmed that some phages, when grown on particular hosts, behaved as complexes of phages. This is most likely because of the presence of inactive phage-related genomes (or their parts) in the bacterial strains which are capable of being reactivated or which can recombine with lytic phages. Furthermore, changes of the bacterial hosts used for maintenance of phages must be avoided as these can drastically modify the parameters of the phage preparations, including host range and lytic activity., Significance and Impact of the Study: This work shows that the optimal host and growth conditions must be carefully studied and selected for the production of each bacteriophage candidate for animal therapy.

For chromosomal localization of the hFVIII human transgene in F2 and F3 generation of transgenic rabbits, FISH-TSA was applied. A short cDNA probe (1250 bp) targeted chromosomes 3, 7, 8, 9 and 18 of an F2 male (animal 1-3-8). Two transgenic offspring (F3) revealed signal positions in chromosome 3 and chromosomes 3 and 7, respectively. Sequencing and structure analysis of the rabbit orthologous gene revealed high similarity to its human counterpart. Part of the sequenced cDNA (1310 bp) served as a probe for FISH-TSA analysis. The rabbit gene was localized in the q arm terminus of the X chromosome. This result is in agreement with reciprocal chromosome painting between the rabbit and the human. The presented FISH-TSA method provides strong signals without any interspecies reactivity.

A single copy gene, mitochondrial malate dehydrogenase 2 (Mdh2), was localized on Xenopus tropicalis chromosomes by fluorescence in situ hybridization coupled with tyramide signal amplification (FISH-TSA). The respective cDNA was cloned and sequenced. The labeled probe hybridized with a subcentromeric region of the long arms of homologous chromosomes 3. Results of comparison of the gene localization with previously mapped X. laevis paralogs strongly suggest a common evolutionary origin of chromosomes 3 and 8 in X. laevis and chromosome 3 in X. tropicalis. This is the first time that a single copy gene has been visualized on X. tropicalis chromosomes. The FISH-TSA method gives a strong signal with a 1-kb labeled probe., (Copyright 2007 S. Karger AG, Basel.)

The aim of this study was to compare the effect of purified GPBoS and commonly used FCS on porcine oocyte maturation and subsequent embryonic development after their parthenogenetic activation. COCs were obtained from dissected follicles and cultured for 18, 24, 30, 36, 42 and 48 h in M-199 medium either with GPBoS or FCS. After 24 h with GPBoS, 91% of oocytes reached MI stage while in the medium supplemented with FCS, only 29% of oocytes reached the same stage (P < 0.05). The majority of oocytes from the FCS group (61%) reached MI stage approximately 6 h later. In the time periods between 36 to 48 h both groups of oocytes reached the same stage of maturation. After 48 h of culture the oocytes with extruded polar bodies were activated by a single electric pulse and then cultured with 4 mM 6-DMAP. Activated oocytes were cultured in PZM-3 medium supplemented with 3 mg/ml of BSA. After 7 days, the development and the quality of embryos were evaluated. The results showed that the maturation of oocytes in the presence of GPBoS significantly increased their subsequent developmental ability when compared with FCS supplementation (27% vs. 19% of blastocysts, P < 0.05). However, differential staining revealed that once blastocysts were formed in either group, they had the same total cell number (40 vs. 41) and also the ICM/total cell ratio (0.27 vs. 0.29).

Fluorescent in situ hybridization followed by tyramide signal amplification was used to map the site of the c-SRC1 gene on XENOPUS LAEVIS chromosomes. Positive results were obtained with a cDNA probe of about 1 kb. The c-SRC1 gene is located in the subcentromeric region in the long arm of one of the acrocentric chromosomes of the G category (classified according to Graf and Kobel, 1991). The c-SRC1 gene and the XENOPUS major histocompatibility complex (MHC) 1b locus, which consists of 20 tandemly arranged gene copies, are situated on different chromosomes of the G category., (Copyright 2003 S. Karger AG, Basel)

A fragment of ME2 cDNA from exon 2 to exon 11 was sequenced and the sequence submitted to GenBank. Analysis of the intron, probably intron 13, revealed a polymorphism which is due to the presence of tandem repetitions of Xstir elements. Genetic analysis of the parents and the offspring showed a standard distribution of intron variants. This distribution was not dependent on sex. We conclude, contrary to previous reports, that the ME2 gene is not linked to sex. Consequently, the Xstir polymorphism can be used as a tool for genetic analysis.

Haemorrhage within the posterior fossa (PF) after supratentorial surgery is a very rare and exceedingly dangerous complication. Only 28 cases were found in the literature. Up to now, no pathogenetic factor has decisively proven to be the cause of this phenomenon. We present clinical details of a patient operated on for aneurysm of the anterior communicating artery. Lumbar drainage was used during surgery, with the loss of a large amount of cerebrospinal fluid (200 ml). Other causes in our case which may have led to cerebellar shift or a critical increase in transmural venous pressure with subsequent vascular disruption and haemorrhage were extreme head rotation during lengthy surgery and blood pressure peaks in the early postoperative period. Repeated computed tomography (CT) allowed immediate diagnosis of this complication and control of its conservative management. After postponed ventriculoperitoneal shunt, the patient recovered completely.

We studied, for the first time to our knowledge, transient stimulated Raman scattering in H(2)- D(2) and H(2)- CH(4) gas mixtures excited with 200-fs and 2-ps, 390-nm pulses. Depending on the composition and partial pressure, of those gases, we observed approximately 40 output lines simultaneously. Under optimum conditions a conversion efficiency of as much as 8% in the combination Raman line at 556 nm was obtained in the H(2)- CH(4) gas mixtures. The difference between picosecond and femtosecond pumping is due to self-phase modulation and to generation of a spectral continuum in the latter. Our calculation of the ratio of the partial pressures is in a good agreement with the experimental results. A new method for measuring the unknown Raman scattering cross section of gas molecules is suggested.

We report measurement of efficient amplification of weak femtosecond supercontinuum seed pulses by use of a noncollinear optical parametric process in BBO crystal pumped with 150-fs pulses from a frequency-doubled regenerative-amplified Ti:sapphire laser at 390nm . The highest amplification factor, 10(8) , was achieved for 3x10(-16)J energy seed pulses at wavelength of 560nm.