Remodeling of the malaria parasite and host human red cell by vesicle amplification that induces artemisinin resistance

Artemisinin resistance threatens worldwide malaria control and elimination. Elevation of phosphatidylinositol-3-phosphate (PI3P) can induce resistance in blood stages of Plasmodium falciparum The parasite unfolded protein response (UPR) has also been implicated as a proteostatic mechanism that may diminish artemisinin-induced toxic proteopathy. How PI3P acts and its connection to the UPR remain unknown, although both are conferred by mutation in P falciparum Kelch13 (K13), the marker of artemisinin resistance. Here we used cryoimmunoelectron microscopy to show that K13 concentrates at PI3P tubules/vesicles of the parasite's endoplasmic reticulum (ER) in infected red cells. K13 colocalizes and copurifies with the major virulence adhesin PfEMP1. The PfEMP1-K13 proteome is comprehensively enriched in multiple proteostasis systems of protein export, quality control, and folding in the ER and cytoplasm and UPR. Synthetic elevation of PI3P that induces resistance in absence of K13 mutation also yields signatures of proteostasis and clinical resistance. These findings imply a key role for PI3P-vesicle amplification as a mechanism of resistance of infected red cells. As validation, the major resistance mutation K13C580Y quantitatively increased PI3P tubules/vesicles, exporting them throughout the parasite and the red cell. Chemical inhibitors and fluorescence microscopy showed that alterations in PfEMP1 export to the red cell and cytoadherence of infected cells to a host endothelial receptor are features of multiple K13 mutants. Together these data suggest that amplified PI3P vesicles disseminate widespread proteostatic capacity that may neutralize artemisinins toxic proteopathy and implicate a role for the host red cell in artemisinin resistance. The mechanistic insights generated will have an impact on malaria drug development

Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli

Against the outdated belief that inclusion bodies (IBs) in Escherichia coli are only inactive aggregates of misfolded protein, and thus should be avoided during recombinant protein production, numerous biopharmaceutically important proteins are currently produced as IBs. To obtain correctly folded, soluble product, IBs have to be processed, namely, harvested, solubilized, and refolded. Several years ago, it was discovered that, depending on cultivation conditions and protein properties, IBs contain partially correctly folded protein structures, which makes IB processing more efficient. Here, we present a method of tailored induction of recombinant protein production in E. coli by a mixed feed system using glucose and lactose and its impact on IB formation. Our method allows tuning of IB amount, IB size, size distribution, and purity, which does not only facilitate IB processing, but is also crucial for potential direct applications of IBs as nanomaterials and biomaterials in regenerative medicine

SMARCB1-deficient sinonasal adenocarcinoma: a rare variant of SWI/SNF-deficient malignancy often misclassified as high-grade non-intestinal-type sinonasal adenocarcinoma or myoepithelial carcinoma

SMARCB1-deficient sinonasal adenocarcinoma is a rare variant of SWI/SNF-deficient malignancies with SMARCB1 loss and adenocarcinoma features. More than 200 high-grade epithelial sinonasal malignancies were retrieved. A total of 14 cases exhibited complete SMARCB1 (INI1) loss and glandular differentiation. SMARCA2 and SMARCA4 were normal, except for one case with a loss of SMARCA2. Next-generation sequencing (NGS) and/or fluorescence in situ hybridization (FISH) revealed an alteration in the SMARCB1 gene in 9/13 cases, while 2/13 were negative. Two tumors harbored SMARCB1 mutations in c.157C > T p.(Arg53Ter) and c.842G > A p.(Trp281Ter). One harbored ARID1B mutations in c.1469G > A p.(Trp490Ter) and MGA c.3724C > T p.(Arg1242Ter). Seven tumors had a SMARCB1 deletion. One carried an ESR1 mutation in c.644-2A  >  T , and another carried a POLE mutation in c.352_374del p.(Ser118GlyfsTer78). One case had a PAX3 mutation in c.44del p.(Gly15AlafsTer95). Histomorphology of SMARCB1-deficient adenocarcinoma was oncocytoid/rhabdoid and glandular, solid, or trabecular in 9/14 cases. Two had basaloid/blue cytoplasm and one showed focal signet ring cells. Yolk sac tumor-like differentiation with Schiller-Duval-like bodies was seen in 6/14 cases, with 2 cases showing exclusively reticular-microcystic yolk sac pattern. Follow-up of a maximum of 26 months (median 10 months) was available for 8/14 patients. Distant metastasis to the lung, liver, mediastinum, bone, and/or retroperitoneum was seen in 4/8 cases. Locoregional failure was seen in 75% of patients, with 6/8 local recurrences and 3 cervical lymph node metastases. At the last follow-up, 5 of 8 (62%) patients had died of their disease 2 to 20 months after diagnosis (median 8.2 months), and 3 were alive with the disease. The original diagnosis was usually high-grade non-intestinal-type adenocarcinoma or high-grade myoepithelial carcinoma. A correct diagnosis of these aggressive tumors could lead to improved targeted therapies with potentially better overall disease-specific survival

Laser radiation in minimally invasive medicine

Introduction. The development trend of modern medicine is minimal invasiveness while maintaining sufficient radicality . This is aided by a great deal of new knowledge, revolutionary technical achievements and work approaches. Examples of this are numerous - an excimer laser for myopia, radio frequency thermoablation in the treatment of liver metastases, the use of a harmonic scalpel in endoscopic or open surgery. These are all examples where minimum damage is incurred on the "way" to a treating an organ with the same radicalism as with traditional surgery. The ultrasonic knife and radiofrequency thermotherapy have entered medicine in a multidisciplinary way, which, however, remains incomparable with the expansit of the phenomenon called laser. Lasers bring together a group of devices with very diverse characteristics, enabling applications in many fields of medicine. Wavelength defined by an active medium, coherence, collimation and monochromaticity predisposes lasers to a single technically unique selective influence of various tissues with their great energy. This enables a device belonging to a group of lasers to crush kidney stones, as well as evaporate a cornea or cut enamel. Objectives of the work. Our objective was to determine the benefits of laser medicine to minimize invasiveness in..

Risk of Postoperative Bleeding in Tonsillectomy for Peritonsillar Abscess, as Opposed to in Recurrent and Chronic Tonsillitis—A Retrospective Study

Tonsillectomy is a routine surgery in otorhinolaryngology and the occurrence of postoperative bleeding is not a rare complication. The aim of this retrospective, observational, analytic, cohort study is to compare the incidence of this complication for the most common indications. A group of patients indicated for tonsillectomies for peritonsillar abscess (group I) was compared to a group of patients indicated for chronic and recurrent tonsillitis (group II). There are a lot of pathophysiological differences in patients indicated for acute tonsillectomy for peritonsillar abscess and in patients indicated for elective tonsillectomy for chronic or recurrent tonsillitis. No technique to minimize the risk of bleeding after tonsillectomy has been found and a large part of postoperative bleeding occurs in postoperative home-care, which makes this issue topical. In total, 2842 unilateral tonsillectomies from the years 2014–2019 were included in the study. Bleeding occurred in 10.03% and, surprisingly, despite completely different conditions in the field of surgery (oedema, acute inflammation in peritonsillar abscess), there was no statistically significant difference between incidence of postoperative bleeding in the studied groups (p = 0.9920). The highest incidence of bleeding was found in the patients of group I on the eighth postoperative day, with those aged 20–24 years (p = 0.0235) being the most at risk, and in group II, on the sixth postoperative day, with those aged 25–29 years (p = 0.0128) and 45–49 years (p = 0.0249) being the most at risk

Non-fiction literature in Czech online media in years 2015 to 2018

This bachelor's thesis focuses on non-fiction literature in Czech online media during years 2015 to 2018. Objective of this thesis is to look if, and how, is non-fiction literature present in Czech online media. To accomplish this objective, analysis of popular Czech digital journalism websites is carried out alongside the use of two popular search engines from Google.cz and Seznam,cz. Besides this main objective, the unique variant of non-fiction literature specific for Czech Republic, the so-called "literatura faktu" or "literature of (a) fact(s)", is better defined both as a concept and as a term. Chapters 1 to 3 are striving to find a better definition of non-fiction literature in Czech Republic, but they also concentrate on history of non-fiction in Czechia and Czechoslovakia and on its differences and similarities with Anglo-Saxon and international non-fiction. Fourth chapter of the thesis focuses on analysis and methodology used to find relevant research results. The fifth, sixth and seventh chapters are thoroughly investigating Czech internet to find results for non-fiction literature, while the eight chapter is summarizing all found information. Along with Summary itself, it puts newfound knowledge in context

Problematika homogenní aplikace laserového záření na rozsáhlé plochy lidské tkáně

Laserové přístroje jsou dnes řešeny mnoha technickými způsoby, které se odvíjí od jejich cílového použití, přičemž velmi speciální využití nacházejí lasery v lékařství. Účinek medicínských laserů je dán jejich základními fyzikálními vlastnostmi (vlnová délka, kolimace, koherence, monochromatičnost) a jejich interakcí s biologickými vlastnostmi cílové tkáně. Specifickou oblastí aplikace laserů v lékařství jsou situace, kde je z klinických důvodů vyžadována aplikace laserového záření na rozsáhlejší plochy (desítky až stovky cm2) při zajištění maximální homogenity distribuce laserové energie. Obsahem tohoto článku je kvantifikovat dostupné metody plošné aplikace nejenom technicky, ale i na základě biologického efektu in vivo s dlouhodobým sledováním. Práce vznikla jako výsledek společného výzkumu LFUK V Plzni a FEL ZČU v Plzni.Laser devices today are built using many different technical designs determined by their target application where a very special use have lasers in medicine. The effects of medical lasers are determined by their basic physical properties (wavelength, collimation, coherence, monochromaticity) and their interactions with biological properties of the target tissue. A specific area of application of lasers in medicine are situations where clinical reasons require the laser irradiation over larger areas (tens to hundreds cm2) at maximum homogeneity of the distribution of laser energy. The aim of this article is to quantify the available methods for surface application not only technically, but also on the basis of biological effects in vivo with long-term monitoring. This work is a result of joint research of Charles University Medical School and University of West Bohemia, Department of Electromechanics and Power Electronics both in Pilsen