Chem. Listy 2022; 116: 101−109. doi: 10.54779/ chl20220101 vladimir.pliska@biol.ethz.ch, parizek@porodnice.cz Došlo 12. 10. 2021, přijato 1. 11. 2021.

Abstract: Licence agreements between the Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Sciences, Prague (IOCB), and the pharmaceutical company Ferring AB Malmö enabled the Swedish company to produce and commercialize worldwide a number of neurohypophyseal peptides designed at the IOCB. Several of them found therapeutic applications. dDAVP: 1-deamino-8-D-arginine-vasopressin was designed in one of the IOCB peptide laboratories (M. Zaoral and F. Šorm) in 1967. It displayed an extremely high antidiuretic activity (various tests indicate a 2- to 50-fold increase, as compared to arginine vasopressin) and a very low pressor activity. The peptide (covered by the U.S. Patent No. 3,497,491, 1970) has been used as a preferred drug in the substitution therapy of the central form of diabetes insipidus (Minirin®, today as Desmopressin INN). Besides, as later discovered (Mannucci et al. 1977), dDAVP increases the plasma concentration of the blood clottiing factor VIII. This fact extended its clinical use as haemostatics in cases of milder forms of haemophilia A, von Willebrand-Jürgens syndrome and some thrombocyte dysfunctions. Despite of the clinical success of dDAVP, a closer look reveals certain inadequacies in the presently available pharmacological data: several reports declare activity values and the prolongation effect (index of persistence) in very broad ranges. Triglycyl-8- -lysine-vasopressin (Terlipressin), a peptide with lysine vasopressin chain extended at the N-terminal by a triglycine residue, acts mainly as a prodrug (releasing lysine vasopressin after aminopeptidase splitting at the Na group). The analogue belongs to the so-called „synthetic hormonogens“; individual peptides carrying various acylating groups were synthesized in the midsixties at the IOCB and legally protected by U.S. Patent No. 3,558,590 (1968). It was a part of the licence agreements mentioned above. The activities of triglycyl-8-lysine-vasopressin (both antidiuretic and vasopressor) are about 100 times lower than those of lysine vasopressin, but its persistence is 5 times longer. As such, it is occasionally used in emergency medicine in cases of esophageal (and other gastroenteral) bleeding, traumatic or septic shock, in cirrhotic patients and patients with portal hypertension. Its use as an early abortion drug was discussed but not pursued. Carbetocin (deaminocarba1-2-O-methyltyrosin-oxytocin) was synthetized in the laboratory of Karel Jošt at the IOCB before 1971; its synthesis was covered by a Czechoslovak patent (CS-149,028 B1) in June 1973 (at that time, Czechoslovak patent law did not provide for the patentability of substances as such) and first published in a biophysical communication by Frič et al., 1974). As a part of the licence agreement, it was included in the production program of Ferring AB, but marketed later also by several other pharmaceutical companies due to an incomplete patent protection. The peptide is a moderately active uterotonic partial agonist and as such has been utilized in veterinary obstetrics for delivery induction in cows and (multiparous) pigs: its milder and better--controlled uterotonic action was found preferential as compared to oxytocin so far used for these purposes. In the last two decades, carbetocin has been commonly used also in the human obstetrics, especially to prevent the peri- and post-partum haemorrhage, from the maternal side the most frequent and most severe delivery complication. It became a life-saving drug in emergency obstetrics.

• Klíčová slova
• carbetocin,
• MeSH
• desmopresin aplikace a dávkování   dějiny   farmakologie   terapeutické užití   MeSH
• lidé MeSH
• neurohypofyzární hormony * dějiny   farmakologie   klasifikace   MeSH
• oxytocin antagonisté a inhibitory   dějiny   farmakologie   MeSH
• poporodní krvácení farmakoterapie   MeSH
• terlipresin aplikace a dávkování   dějiny   farmakologie   terapeutické užití   MeSH
• uterotonika aplikace a dávkování   dějiny   farmakologie   klasifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH
• Geografické názvy
• Česká republika MeSH
• Švédsko MeSH

Colorectal cancer is influenced by genetic mutations, lifestyle factors, and diet, particularly high fat intake, which raises bile acid levels in the intestinal lumen. This study hypothesized that bile acids contribute to tumorigenesis by disrupting ion transport and ATPase activity in the intestinal mucosa. The effects of 3-sulfo-taurolithocholic acid (TLC-S) on ATPase activity were investigated in colorectal cancer samples from 10 patients, using adjacent healthy tissue as controls, and in rodent liver function. ATPase activity was measured spectrophotometrically by determining inorganic phosphorus (Pi) in postmitochondrial fractions. Ca2+ dynamics were assessed in isolated mouse hepatocytes with fluorescence imaging, and rat liver mitochondria were studied using polarographic methods to evaluate respiration and oxidative phosphorylation. TLC-S increased Na+/K+ ATPase activity by 1.5 times in colorectal cancer samples compared to controls (p ≤ 0.05). In healthy mucosa, TLC-S decreased Mg2+ ATPase activity by 3.6 times (p ≤ 0.05), while Mg2+ ATPase activity in cancer tissue remained unchanged. TLC-S had no significant effect on Ca2+ ATPase activity in healthy colon mucosa but showed a trend toward decreased activity in cancer tissue. In rat liver, TLC-S decreased Ca2+ ATPase and Na+/K+ ATPase activities while increasing basal Mg2+ ATPase activity (p ≤ 0.05). Additionally, TLC-S induced cytosolic Ca2+ signals in mouse hepatocytes, partially attenuated by NED-19, an NAADP antagonist (p ≤ 0.05). TLC-S also reduced the V3 respiration rate of isolated rat liver mitochondria during α-ketoglutarate oxidation. These findings suggest that TLC-S modulates ATPase activity differently in cancerous and healthy colon tissues, playing a role in colorectal cancer development. In rat liver, TLC-S affects mitochondrial activity and ATPase function, contributing to altered cytosolic calcium levels, providing insight into the mechanistic effects of bile acids on colorectal cancer and liver function.

• Publikační typ
• časopisecké články MeSH

• MeSH
• věda MeSH
• veterinární lékařství MeSH
• Publikační typ
• rozhovory MeSH

The global threat of neglected tropical diseases (NTDs) constitutes a public health issue in underdeveloped countries. Zoonotic helminthiases are the most common human NTD agents in developing countries in sub-Saharan Africa, Asia, and the Americas, causing a global burden of disease that exceeds that of more recognized infectious diseases such as malaria and tuberculosis. Wild canids are well-known mammals that act as natural reservoirs of zoonotic-relevant helminthiasis worldwide, thus playing a pivotal role in their epidemiology and transmission to humans. Here we evaluate the occurrence of zoonotic gastrointestinal helminths in two Neotropical wild canid species from the Amazonian and Andean regions of Colombia, i.e., the bush dog (Speothos venaticus) and the crab-eating fox (Cerdocyon thous). We recovered tapeworm proglottids from bush dog fecal samples and identified them molecularly as the canine-specific lineage of Dipylidium caninum by using cytochrome c oxidase subunit I (cox1) gene sequences. Moreover, examination of a crab-eating fox during necropsy revealed the presence of non-embryonated eggs of the neglected nematode Lagochilascaris cf. minor, in addition to eggs and gravid proglottids of the cestode Spirometra mansoni. These findings represent the first report of zoonotic-relevant cestodes, i.e., D. caninum ("canine genotype"), S. mansoni, and the nematode L. cf. minor, in bush dogs and crab-eating foxes as final hosts. The occurrence of these zoonotic helminthiases in wild canid species calls for regular monitoring programs to better understand the epidemiology and transmission routes of neglected dipylidiasis, lagochilascariosis, and sparganosis in South America.

• Publikační typ
• časopisecké články MeSH

Species of Sarcocystis use various vertebrates as intermediate or definitive hosts in their life cycles. One of these is snakes, whose role as intermediate hosts for these protozoans is scarce; in fact, there are six records, but only three with molecular characterization. An imported green tree python was involved in the morphological and molecular characterization (four loci) of a new species of Sarcocystis localized in skeletal muscles. Sarcocystis moreliae sp. nov. has a type 1 sarcocyst with a smooth wall and is genetically similar (at the 18S rRNA gene) to two unnamed species of Sarcocystis found in Lytorhynchus diadema from Oman and Varanus salvator macromaculatus from Malaysia, but their detailed comparison is impossible. The new species showed lower similarity to its congeners in other loci (28S rRNA, ITS1, and cox1). This is the first morphological and genetic characterization of a Sarcocystis species in snakes of the genus Morelia, particularly M. viridis, using four loci, but more data are needed to fill the knowledge gap about snakes as intermediate hosts of Sarcocystis.

• Publikační typ
• časopisecké články MeSH

Tick-transmitted apicomplexans of the genera Cytauxzoon and Hepatozoon affect a wide range of felids worldwide, but little is known about them. Recently, several studies addressed the species circulating in Europe, their distribution, and their hosts. Molecular assays are the method of choice for their detection. Unfortunately, conventional PCRs already described are time- and cost-consuming and specific for either Hepatozoon or Cytauxzoon detection. This study was developed to evaluate (i) the occurrence of Cytauxzoon and Hepatozoon in felids using a fast and cost-saving real-time PCR capable of detecting both protozoa simultaneously, (ii) the distribution of Cytauxzoon and Hepatozoon species in north-eastern Italy, and (iii) the involvement of other susceptible felid hosts in the same area. An SYBR® Green-based real-time PCR with primers targeting the 18S-rRNA was validated and applied to 237 felid samples, i.e., whole blood from 206 domestic cats and 12 captive exotic felids, and tissues from 19 wildcats. Positive results were obtained by melting temperature curve analysis due to the specific melting peak (i.e., 81°C Cytauxzoon spp.; 78-78.5°C Hepatozoon spp.). Positive samples were subjected to conventional PCR, followed by sequencing for species identification. Phylogenetic analyses were performed to assess relatedness among European isolates. Data on domestic cats (age class, sex, origin, management, and lifestyle) were recorded, and statistical analyses were performed to identify potential risk factors. A total of 31 (15%) domestic cats were positive for Hepatozoon spp. (i.e., 12 for H. felis, 19 for H. silvestris), while six (2.9%) for C. europaeus. The prevalence of Hepatozoon felis was significantly (p < 0.05) higher in domestic cats, while H. silvestris was higher in strays and animals from the Eastern region (i.e., Friuli-Venezia Giulia). Cytauxzoon europaeus was detected only in stray cats from Friuli-Venezia Giulia (province of Trieste). Among captive felids, one tiger was infected with H. felis and another with H. silvestris; eight out of 19 (42%) wildcats were positive for Hepatozoon spp. (i.e., six with H. felis, two with H. silvestris) and four out of 19 (21%) for Cytauxzoon europaeus. Outdoor lifestyle and origin (i.e., Friuli-Venezia Giulia region) were the most relevant risk factors for H. silvestris and C. europeus infections. Conversely, H. felis was most frequently isolated from domestic cats, suggesting different modes of transmission.

• Publikační typ
• časopisecké články MeSH

Intervertebral disc extrusion (IVDE) is a common neurological condition in many dog breeds. This study aimed to describe this condition in Yorkshire terriers (YT) and calculate the prevalence of this condition amongst the YTs with neurological diseases. This is a double-centre retrospective study which was conducted in two arms. The first part of the study, describing the clinical features and prognosis of cervical (C) IVDE in YTs, is based on data from 2005 to 2021. The second part of the study calculated the prevalence of C IVDE amongst the YTs with neurological diseases based on data from 2016 to 2021. A retrospective search through the medical records was conducted. YTs with C IVDE diagnosed with MRI and confirmed surgically were eligible for inclusion in this study. Sixty YTs were included in the first part of the study. There were 48 (80%) dogs with acute onset and 12 (20%) with chronic onset with acute deterioration. Ambulation was preserved in 31 (51.7%) dogs on admission, and the remaining 29 (48.3%) dogs were non-ambulatory. No significant association was found between ambulation on admission and recovery status (p = 0.547). Seventy-three intervertebral spaces were treated during the surgical intervention. Relapses were seen in seven (11.7%) dogs. Forty-nine (81.7%) dogs were ambulatory at discharge. A complete recovery was observed in 46 (76.7%) dogs; the remaining dogs (14, 23.3%) were classified as incomplete recovery. A significant difference was found in time to ambulation (p = 0.0238) and time to discharge (p = 0.0139) between the on-admission ambulatory and non-ambulatory dogs. Three hundred and eight YTs were diagnosed with neurological diseases between 2016 and 2021 in one referral centre. C IVDE was diagnosed in 31 (10.06%) dogs. This is the first study explicitly describing the C IVDE in YTs and establishing the prevalence of this condition amongst YTs with other neurological disorders.

• Publikační typ
• časopisecké články MeSH

Bovine papillomavirus type 1 L1 protein was produced in a baculovirus expression system and purified as virus-like particles (VLPs) by affinity chromatography using lectins. The morphological integrity of VLPs was confirmed by electron microscopy. Differences between the two detected variants were deciphered by mass spectrometry of peptides (MALDI-TOF). Mice were immunized with purified VLPs in doses of 10, 25, or 50 μg in combination with 1% saponin and 15% alhydrogel per dose as adjuvants. Analysis of the humoral immune response revealed increased levels of specific antibodies detected 3 weeks after the first immunization in all groups of animals. This was further significantly increased by the booster applied 3 weeks after the first dose, with the best immune response in a group of mice immunized by the largest dose of antigen. BPV1 L1 VLPs purified by affinity chromatography using lectins could be used for prophylactic immunization in veterinary medicine.

• Publikační typ
• časopisecké články MeSH

OBJECTIVE: The aim of this systematic review is to summarize outcomes of studies focused on the effects of opioids, injectable sedative and anaesthetic drugs and inhalant anaesthetics on tear production in dogs. This manuscript complements the systematic review describing the effect of anaesthetics on intraocular pressure in dogs (Pierce-Tomlin et al. 2020). Databases used A detailed search of scientific references has been performed. PubMed, Web of Science, Science Direct and Google Scholar databases were used to search for sources using free text terms 'Dog' or 'Canine', 'Anaesthesia' or 'Anaesthetic' or 'Sedative' or 'Opioid' or the name of used opioids, sedative and anaesthetic drugs and 'Tear' or 'Schirmer' or 'Lacrimation'. The time frame searched was from 1960 to October 2021. Any published manuscripts that were concerned with sedative or anaesthetic drugs administered systemically in the dog and tear production were evaluated. CONCLUSIONS: Low doses of α2-adrenoceptor agonists, neuroleptics, benzodiazepines, opioids, propofol or alfaxalone administered alone have no clinically significant effect on aqueous tear production in healthy dogs measured by the Schirmer tear test I (STT-I). Intramuscular injection of ketamine increases STT-I values. Higher doses of α2-adrenoceptor agonists and combinations of anaesthetics, including inhaled anaesthetics, always clinically significantly decrease tear production.

• MeSH
• adrenergní receptory MeSH
• anestetika * farmakologie   MeSH
• hypnotika a sedativa farmakologie   MeSH
• propofol * farmakologie   MeSH
• psi MeSH
• slzy MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• systematický přehled MeSH

BACKGROUND: Current SARS-CoV-2 detection platforms lack the ability to differentiate among variants of concern (VOCs) in an efficient manner. CRISPR/Cas (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated) based detection systems have the potential to transform the landscape of COVID-19 diagnostics due to their programmability; however, most of these methods are reliant on either a multi-step process involving amplification or elaborate guide RNA designs. METHODS: Three Cas12b proteins from Alicyclobacillus acidoterrestris (AacCas12b), Alicyclobacillus acidiphilus (AapCas12b), and Brevibacillus sp. SYP-B805 (BrCas12b) were expressed and purified, and their thermostability was characterised by differential scanning fluorimetry, cis-, and trans-cleavage activities over a range of temperatures. The BrCas12b was then incorporated into a reverse transcription loop-mediated isothermal amplification (RT-LAMP)-based one-pot reaction system, coined CRISPR-SPADE (CRISPR Single Pot Assay for Detecting Emerging VOCs). FINDINGS: Here we describe a complete one-pot detection reaction using a thermostable Cas12b effector endonuclease from Brevibacillus sp. to overcome these challenges detecting and discriminating SARS-CoV-2 VOCs in clinical samples. CRISPR-SPADE was then applied for discriminating SARS-CoV-2 VOCs, including Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), and Omicron (B.1.1.529) and validated in 208 clinical samples. CRISPR-SPADE achieved 92·8% sensitivity, 99·4% specificity, and 96·7% accuracy within 10-30 min for discriminating the SARS-CoV-2 VOCs, in agreement with S gene sequencing, achieving a positive and negative predictive value of 99·1% and 95·1%, respectively. Interestingly, for samples with high viral load (Ct value ≤ 30), 100% accuracy and sensitivity were attained. To facilitate dissemination and global implementation of the assay, a lyophilised version of one-pot CRISPR-SPADE reagents was developed and combined with an in-house portable multiplexing device capable of interpreting two orthogonal fluorescence signals. INTERPRETATION: This technology enables real-time monitoring of RT-LAMP-mediated amplification and CRISPR-based reactions at a fraction of the cost of a qPCR system. The thermostable Brevibacillus sp. Cas12b offers relaxed primer design for accurately detecting SARS-CoV-2 VOCs in a simple and robust one-pot assay. The lyophilised reagents and simple instrumentation further enable rapid deployable point-of-care diagnostics that can be easily expanded beyond COVID-19. FUNDING: This project was funded in part by the United States-India Science & Technology Endowment Fund- COVIDI/247/2020 (P.K.J.), Florida Breast Cancer Foundation- AGR00018466 (P.K.J.), National Institutes of Health- NIAID 1R21AI156321-01 (P.K.J.), Centers for Disease Control and Prevention- U01GH002338 (R.R.D., J.A.L., & P.K.J.), University of Florida, Herbert Wertheim College of Engineering (P.K.J.), University of Florida Vice President Office of Research and CTSI seed funds (M.S.), and University of Florida College of Veterinary Medicine and Emerging Pathogens Institute (R.R.D.).

• MeSH
• Brevibacillus * genetika   MeSH
• COVID-19 * diagnóza   MeSH
• guide RNA, Kinetoplastida MeSH
• lidé MeSH
• SARS-CoV-2 genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH