PNA6, a Lactosyl Analogue of Angiotensin-(1-7), Reverses Pain Induced in Murine Models of Inflammation, Chemotherapy-Induced Peripheral Neuropathy, and Metastatic Bone Disease.

Pain is the most significant impairment and debilitating challenge for patients with bone metastasis. Therefore, the primary objective of current therapy is to mitigate and prevent the persistence of pain. Thus, cancer-induced bone pain is described as a multifaceted form of discomfort encompassing both inflammatory and neuropathic elements. We have developed a novel non-addictive pain therapeutic, PNA6, that is a derivative of the peptide Angiotensin-(1-7) and binds the Mas receptor to decrease inflammation-related cancer pain. In the present study, we provide evidence that PNA6 attenuates inflammatory, chemotherapy-induced peripheral neuropathy (CIPN) and cancer pain confined to the long bones, exhibiting longer-lasting efficacious therapeutic effects. PNA6, Asp-Arg-Val-Tyr-Ile-His-Ser-(O-ß-Lact)-amide, was successfully synthesized using solid phase peptide synthesis (SPPS). PNA6 significantly reversed inflammatory pain induced by 2% carrageenan in mice. A second murine model of platinum drug-induced painful peripheral neuropathy was established using oxaliplatin. Mice in the oxaliplatin-vehicle treatment groups demonstrated significant mechanical allodynia compared to the oxaliplatin-PNA6 treatment group mice. In a third study modeling a complex pain state, E0771 breast adenocarcinoma cells were implanted into the femur of female C57BL/6J wild-type mice to induce cancer-induced bone pain (CIBP). Both acute and chronic dosing of PNA6 significantly reduced the spontaneous pain behaviors associated with CIBP. These data suggest that PNA6 is a viable lead candidate for treating chronic inflammatory and complex neuropathic pain.

Antifungal Activity of Glucosinolate-Derived Nitriles and Their Synergistic Activity with Glucosinolate-Derived Isothiocyanates Distinguishes Various Taxa of Brassicaceae Endophytes and Soil Fungi.

The glucosinolates of Brassicaceae plants are converted into bioactive isothiocyanates and other volatiles during a challenge by pathogens and other biotic stressors. However, the role of alternative downstream products with weaker potency (e.g., nitriles) is far from being fully understood. This study tested the possible synergistic antifungal interaction between various glucosinolate-derived nitriles and 2-phenylethyl isothiocyanate (PEITC) on 45 fungal strains, including endophytes from horseradish roots (Brassicaceae) and soil fungi, using an airtight system enabling the accurate study of extremely volatile antifungal agents. The median minimal inhibitory concentrations (MICs) were 1.28, 6.10, 27.00 and 49.72 mM for 1H-indole-3-acetonitrile (IAN), 3-phenylpropanenitrile (PPN), 4-(methylsulfanyl)-butanenitrile (MSBN) and 3-butenenitrile (BN, = allyl cyanide), respectively. Thus, nitriles were considerably weaker antifungal agents compared to PEITC with a median MIC of 0.04 mM. For the same nitriles, the median fractional inhibitory concentration indices (FICIs) of the combinations were 0.562, 0.531, 0.562 and 0.625, respectively. Altogether, 47.7%, 56.8%, 50.0% and 27.3% of tested fungal strains showed a synergistic antifungal activity (FICI ≤ 0.5) for the nitrile-isothiocyanate combinations, respectively. Hypocreales strains showed the least sensitivity towards the GSL decomposition products and their combinations. The mean MIC values for PEITC showed 0.0679 ± 0.0358, 0.0400 ± 0.0214, 0.0319 ± 0.0087 and 0.0178 ± 0.0171 mM for Hypocreales, Eurotiales, Glomerellales and Pleosporales, respectively. In addition, nitriles, especially IAN, also showed significant differences. For the same fungi, the median FICI values fell in the ranges of 0.61-0.67, 0.52-0.61, 0.40-0.50 and 0.48-0.67, respectively, depending on the nitrile. Our results suggest that glucosinolate-derived nitriles may enhance isothiocyanate antifungal activity and that they may play an active role in shaping the plant microbiome and contribute to the filtering of microbes by plants.

Disruption of the Schizosaccharomyces japonicus lig4 Disturbs Several Cellular Processes and Leads to a Pleiotropic Phenotype.

Gene targeting is a commonly used method to reveal the function of genes. Although it is an attractive tool for molecular studies, it can frequently be a challenge because its efficiency can be low and it requires the screening of a large number of transformants. Generally, these problems originate from the elevated level of ectopic integration caused by non-homologous DNA end joining (NHEJ). To eliminate this problem, NHEJ-related genes are frequently deleted or disrupted. Although these manipulations can improve gene targeting, the phenotype of the mutant strains raised the question of whether mutations have side effects. The aim of this study was to disrupt the lig4 gene in the dimorphic fission yeast, S. japonicus, and investigate the phenotypic changes of the mutant strain. The mutant cells have shown various phenotypic changes, such as increased sporulation on complete medium, decreased hyphal growth, faster chronological aging, and higher sensitivity to heat shock, UV light, and caffeine. In addition, higher flocculation capacity has been observed, especially at lower sugar concentrations. These changes were supported by transcriptional profiling. Many genes belonging to metabolic and transport processes, cell division, or signaling had altered mRNA levels compared to the control strain. Although the disruption improved the gene targeting, we assume that the lig4 inactivation can cause unexpected physiological side effects, and we have to be very careful with the manipulations of the NHEJ-related genes. To reveal the exact mechanisms behind these changes, further investigations are required.

Characterization of Zygosaccharomyces lentus Yeast in Hungarian Botrytized Wines.

Tokaj botrytized sweet wines are traditionally aged for several years in wood barrels or bottles. As they have significant residual sugar content, they are exposed to microbial contamination during ageing. Osmotolerant wine-spoilage yeasts are most commonly found in the Tokaj wine-growing region in the species Starmerella spp. and Zygosaccharomyces spp. For the first time, Z. lentus yeasts were isolated from post-fermented botrytized wines. Our physiological studies confirmed that these yeast strains are osmotolerant, with high sulphur tolerance and 8% v/v alcohol tolerance, and that they grow well at cellar temperature in acidic conditions. Low ß-glucosidase and sulphite reductase activities were observed, whereas protease, cellulase, and α-arabinofuranosidase extracellular enzyme activities were not detected. Molecular biology analyses carried out by RFLP analysis of mtDNA revealed no remarkable differences between strains, while microsatellite-primed-PCR fingerprinting of the (GTG)5 microsatellite and examination of chromosomal pattern revealed considerable diversity. The fermentative vigour of the tested Z. lentus strains was found to be significantly lower compared to the control Saccharomyces cerevisiae (Lalvin EC1118). It can be concluded that Z. lentus is a potential spoilage yeast in oenology which may be responsible for the initiation of secondary fermentation of wines during ageing.

Structure-Based Design of Glycosylated Oxytocin Analogues with Improved Selectivity and Antinociceptive Activity.

Acute and chronic pain is often treated with opioids despite the negative side effects of constipation, physical dependence, respiratory depression, and overdose. The misuse of opioid analgesics has given rise to the opioid crisis/epidemic, and alternate nonaddictive analgesics are urgently needed. Oxytocin, a pituitary hormone, is an alternative to the small molecule treatments available and has been used as an analgesic as well as for the treatment and prevention of opioid use disorder (OUD). Clinical implementation is limited by its poor pharmacokinetic profile, a result of the labile disulfide bond between two cysteine residues in the native sequence. Stable brain penetrant oxytocin analogues have been synthesized by replacement of the disulfide bond with a stable lactam and glycosidation of the C-terminus. These analogues show exquisite selectivity for the oxytocin receptor and potent in vivo antinociception in mice following peripheral (i.v.) administration, supporting further study of their clinical potential.

Correlations Between the Metabolome and the Endophytic Fungal Metagenome Suggests Importance of Various Metabolite Classes in Community Assembly in Horseradish (Armoracia rusticana, Brassicaceae) Roots.

The plant microbiome is an increasingly intensive research area, with significance in agriculture, general plant health, and production of bioactive natural products. Correlations between the fungal endophytic communities and plant chemistry can provide insight into these interactions, and suggest key contributors on both the chemical and fungal side. In this study, roots of various horseradish (Armoracia rusticana) accessions grown under the same conditions were sampled in two consecutive years and chemically characterized using a quality controlled, untargeted metabolomics approach by LC-ESI-MS/MS. Sinigrin, gluconasturtiin, glucoiberin, and glucobrassicin were also quantified. Thereafter, a subset of roots from eight accessions (n = 64) with considerable chemical variability was assessed for their endophytic fungal community, using an ITS2 amplicon-based metagenomic approach using a custom primer with high coverage on fungi, but no amplification of host internal transcribed spacer (ITS). A set of 335 chemical features, including putatively identified flavonoids, phospholipids, peptides, amino acid derivatives, indolic phytoalexins, a glucosinolate, and a glucosinolate downstream product was detected. Major taxa in horseradish roots belonged to Cantharellales, Glomerellales, Hypocreales, Pleosporales, Saccharomycetales, and Sordariales. Most abundant genera included typical endophytes such as Plectosphaerella, Thanatephorus, Podospora, Monosporascus, Exophiala, and Setophoma. A surprising dominance of single taxa was observed for many samples. In summary, 35.23% of reads of the plant endophytic fungal microbiome correlated with changes in the plant metabolome. While the concentration of flavonoid kaempferol glycosides positively correlated with the abundance of many fungal strains, many compounds showed negative correlations with fungi including indolic phytoalexins, a putative glucosinolate but not major glucosinolates and a glutathione isothiocyanate adduct. The latter is likely an in vivo glucosinolate decomposition product important in fungal arrest. Our results show the potency of the untargeted metabolomics approach in deciphering plant-microbe interactions and depicts a complex array of various metabolite classes in shaping the endophytic fungal community.

Design and Synthesis of Brain Penetrant Glycopeptide Analogues of PACAP With Neuroprotective Potential for Traumatic Brain Injury and Parkinsonism.

There is an unmet clinical need for curative therapies to treat neurodegenerative disorders. Most mainstay treatments currently on the market only alleviate specific symptoms and do not reverse disease progression. The Pituitary adenylate cyclase-activating polypeptide (PACAP), an endogenous neuropeptide hormone, has been extensively studied as a potential regenerative therapeutic. PACAP is widely distributed in the central nervous system (CNS) and exerts its neuroprotective and neurotrophic effects via the related Class B GPCRs PAC1, VPAC1, and VPAC2, at which the hormone shows roughly equal activity. Vasoactive intestinal peptide (VIP) also activates these receptors, and this close analogue of PACAP has also shown to promote neuronal survival in various animal models of acute and progressive neurodegenerative diseases. However, PACAP's poor pharmacokinetic profile (non-linear PK/PD), and more importantly its limited blood-brain barrier (BBB) permeability has hampered development of this peptide as a therapeutic. We have demonstrated that glycosylation of PACAP and related peptides promotes penetration of the BBB and improves PK properties while retaining efficacy and potency in the low nanomolar range at its target receptors. Furthermore, judicious structure-activity relationship (SAR) studies revealed key motifs that can be modulated to afford compounds with diverse selectivity profiles. Most importantly, we have demonstrated that select PACAP glycopeptide analogues (2LS80Mel and 2LS98Lac) exert potent neuroprotective effects and anti-inflammatory activity in animal models of traumatic brain injury and in a mild-toxin lesion model of Parkinson's disease, highlighting glycosylation as a viable strategy for converting endogenous peptides into robust and efficacious drug candidates.

Implementation of evidence-based health policy in the field of integrated care: A Ritka Betegségek Nemzeti Eroforrás Központjának tervezési folyamata / Bizonyítékokon alapuló egészségpolitikai döntéshozatal az integrált ellátásban: A Ritka Betegségek Nemzeti Eroforrás Központjának tervezési folyamata

Összefoglaló. Bevezetés: A ritka betegséggel élok ellátásában fontos elorelépések történtek az elmúlt években. Egy következo lépés lehetne hazánkban a Ritka Betegségek Nemzeti Eroforrás (Uni-Versum) Központjának (a továbbiakban: Központ) létrehozása, amely az egészségügyi, szociális és köznevelési szolgáltatásokat összehangolva és kiegészítve személyközpontú ellátást nyújtana a betegek és támogatóik részére. Célkituzés: Célunk az volt, hogy egy nemzetközi tudományos módszertan alapján javaslatokat tegyünk arra, hogy milyen eszközökkel lehet feloldani a Központ megvalósításának lehetséges korlátozó tényezoit. Módszer: A Központ megvalósíthatóságának értékelésére interdiszciplináris szakmai egyeztetést szerveztünk különbözo érintett érdekcsoportok részvételével, a SELFIE H2020 kutatási projekt által kidolgozott módszertan alapján. Az elozetesen rangsorolt legfontosabb korlátozó tényezokre lehetséges megoldási javaslatokat tettünk. Eredmények: A lehetséges korlátozó tényezoket a résztvevok relevánsnak tartották a Központ létrehozásával kapcsolatban, és ezekre összesen 17 olyan konkrét javaslat született, amelyben a résztvevok között egyetértés alakult ki. A javaslatok kiterjedtek az ellátás tartalmára, az alkalmazott technológiák támogató szerepére, a humáneroforrás-korlátok megoldására, a hatékony vezetés és szervezés kialakítására, az összetett finanszírozási struktúra kialakítására és a kutatási lehetoségek megteremtésére is. Megbeszélés: A Központ megvalósítása esetén a ritka betegséggel élok ellátása az egészségügyi, szociális és köznevelési tevékenységeket integráló megközelítés felé mozdulna el. A kutatás során megfogalmazott javaslatok hozzájárulhatnak a Központ létrehozásához, amennyiben megvan az ehhez szükséges szakpolitikai támogatás is. Ezen túlmutatóan, a leírt munkamódszer más integrált ellátási modellek bevezethetoségének elemzéséhez is mintaként szolgálhat. Következtetés: Összefoglalva megállapíthatjuk, hogy a Központ létrehozásához számos, elozetesen is látható korlátozó tényezot kell feloldani. Az érdekcsoportok közös javaslatai alapján kialakítható egy olyan muködési forma, amely az ellátórendszerek kiegészítésével és összehangolásával jelentos társadalmi értéktöbbletet eredményezhet. Orv Hetil. 2021; 162(45): 1818-1825. INTRODUCTION: In Hungary, significant achievements have been made in the care of patients with rare diseases in recent years. A next step could be the establishment of the National Resource Centre for Rare Diseases (hereinafter: Centre) to facilitate patient-centered complex care by the integration and supplementation of existing health, social and educational services. OBJECTIVE: This research aimed to develop recommendations based on international scientific methodology to overcome potential implementation barriers of the aforementioned Centre. METHOD: To evaluate the feasibility of the implementation, we organized an interdisciplinary workshop with representatives of different stakeholder groups, adopting the methodology developed in the SELFIE H2020 research project. During the workshop, we discussed the previously ranked, most significant implementation barriers and made recommendations for potential solutions. RESULTS: The potential implementation barriers were considered relevant by the participants and, reflecting on these barriers, altogether 17 recommendations were developed by consensus. These recommendations were related to the content of service delivery, use of supportive technologies, overcoming workforce issues, establishing effective leadership, implementing a complex financing structure and creating research opportunities. DISCUSSION: Implementation of the Centre would shift the care of rare diseases towards personalized and integrated health, social and educational services. Our recommendations will contribute to the establishment of the Centre, subject to positive policy decision. Furthermore, our methodological approach could support the feasibility assessment of future integrated care solutions and programs. CONCLUSION: Several predictable barriers must be overcome to establish the Centre. Recommendations developed by representatives of relevant stakeholders could support successful implementation and societal value generation. Orv Hetil. 2021; 162(45): 1818-1825.

Genome Comparisons of the Fission Yeasts Reveal Ancient Collinear Loci Maintained by Natural Selection.

Fission yeasts have a unique life history and exhibit distinct evolutionary patterns from other yeasts. Besides, the species demonstrate stable genome structures despite the relatively fast evolution of their genomic sequences. To reveal what could be the reason for that, comparative genomic analyses were carried out. Our results provided evidence that the structural and sequence evolution of the fission yeasts were correlated. Moreover, we revealed ancestral locally collinear blocks (aLCBs), which could have been inherited from their last common ancestor. These aLCBs proved to be the most conserved regions of the genomes as the aLCBs contain almost eight genes/blocks on average in the same orientation and order across the species. Gene order of the aLCBs is mainly fission-yeast-specific but supports the idea of filamentous ancestors. Nevertheless, the sequences and gene structures within the aLCBs are as mutable as any sequences in other parts of the genomes. Although genes of certain Gene Ontology (GO) categories tend to cluster at the aLCBs, those GO enrichments are not related to biological functions or high co-expression rates, they are, rather, determined by the density of essential genes and Rec12 cleavage sites. These data and our simulations indicated that aLCBs might not only be remnants of ancestral gene order but are also maintained by natural selection.

Molecular and comparative genomic analyses reveal evolutionarily conserved and unique features of the Schizosaccharomyces japonicus mycelial growth and the underlying genomic changes.

Fungal pathogens, from phytopathogenic fungus to human pathogens, are able to alternate between the yeast-like form and filamentous forms. This morphological transition (dimorphism) is in close connection with their pathogenic lifestyles and with their responses to changing environmental conditions. The mechanisms governing these morphogenetic conversions are still not fully understood. Therefore, we studied the filamentous growth of the less-known, non-pathogenic dimorphic fission yeast, S. japonicus, which belongs to an ancient and early evolved branch of the Ascomycota. Its RNA sequencing revealed that several hundred genes were up- or down-regulated in the hyphae compared to the yeast-phase cells. These genes belonged to different GO categories, confirming that mycelial growth is a rather complex process. The genes of transport- and metabolic processes appeared especially in high numbers among them. High expression of genes involved in glycolysis and ethanol production was found in the hyphae, while other results pointed to the regulatory role of the protein kinase A (PKA) pathway. The homologues of 49 S. japonicus filament-associated genes were found by sequence alignments also in seven distantly related dimorphic and filamentous species. The comparative genomic analyses between S. japonicus and the closely related but non-dimorphic S. pombe shed some light on the differences in their genomes. All these data can contribute to a better understanding of hyphal growth and those genomic rearrangements that underlie it.

Incidence, Predictive Factors, Clinical Characteristics and Outcome of Non-variceal Upper Gastrointestinal Bleeding - A Prospective Population-based Study from Hungary.

BACKGROUND AND AIMS: Acute non-variceal upper gastrointestinal bleeding (UGIB) is associated with significant morbidity and mortality. Our aim was to evaluate the incidence, management, risk factors and outcomes of acute non-variceal UGIB in a population-based study from Hungary. METHODS: The present prospective one-year study involved six major community hospitals in Western Hungary covering a population of 1,263,365 persons between January 1 and December 31, 2016. Data collection included demographics, comorbidities endoscopic management, Glasgow-Blatchford score (GBS), Rockall score (RS) transfusion requirements, length of hospital stay and mortality. RESULTS: 688 cases of acute non-variceal UGIB were included with an incidence rate of 54.4 (95%CI: 50.5-58.6) per 100,000 per year. Endoscopy was performed within 12 hours in 71.8%. 5.3% of the patients required surgical treatment and the overall mortality was 13.5%. Weekend presentation was associated with increased transfusion requirements (p=0.047), surgery (p=0.016) and mortality (p=0.021). Presentation with hemodynamic instability or presence of comorbidities was associated with transfusion (p<0.001 both), second look endoscopy (p<0.001 both), re-bleeding (p<0.001 both), longer in-hospital stay (p<0.001 both) and mortality (p=0.017 and p<0.001). GBS was associated with transfusion requirement (AUC:0.82; cut-off: GBS >7points), while mortality was best predicted by the post-endoscopic RS (AUC:0.75; cut-off: RS >5points). CONCLUSIONS: Incidence rates of acute non-variceal UGIB in Western Hungary are in line with international trends. Longer pre-hospital time, comorbidities, hemodynamic instability, weekend presentation, treatment with anticoagulants or non-steroidal anti-inflammatory drugs was associated with worse outcomes.

Outcomes of variceal upper gastrointestinal bleeding: A prospective, multicenter, population-based study from West Hungary / A varixeredetu felso gastrointestinalis vérzések elemzése: yugat-magyarországi prospektív, multicentrikus, népességalapú vizsgálat

Összefoglaló. Bevezetés: Az akut varixeredetu gastrointestinalis vérzés napjainkban is jelentos morbiditással és mortalitással jár. Célkituzés: Célunk az akut varixeredetu felso gastrointestinalis vérzések incidenciájának, ellátási folyamatainak és kimeneteli tényezoinek átfogó felmérése volt. Módszer: Prospektív, multicentrikus vizsgálatunk keretében hat nyugat-magyarországi gasztroenterológiai centrum bevonásával elemeztük az ott diagnosztizált és kezelt, varixvérzo betegek adatait. Rögzítettük a demográfiai, az anamnesztikus, a diagnosztikus, valamint a terápiát és a betegség kimenetelét érinto adatokat. Minden beteg esetében kockázat- és predikcióbecslést végeztünk a Glasgow-Blatchford Score (GBS), a pre- és posztendoszkópos Rockall Score (RS) és az American Society of Anesthesiologists (ASA) Score alapján. Eredmények: A vizsgált egyéves periódusban (2016. 01. 01. és 2016. 12. 31. között) 108, akut varixeredetu gastrointestinalis vérzést találtunk (átlagéletkor: 59,6 év). Endoszkópos terápiára 57,4%-ban került sor, 39,8% sclerotherapiában, 18,5% ligatióban részesült. Transzfúziót a betegek 76,9%-a igényelt. A teljes halálozás 24,1% volt. A transzfúziós igény vonatkozásában a legmagasabb prediktív értéku a GBS volt (AUC: 0,793; cut-off: GBS >8 pont). Az ASA-pontszám szignifikáns összefüggést mutatott a transzfúzió-szükséglettel (OR 7,6 [CI 95% 2,7-21,6]; p<0,001), az endoszkópos intervencióval (OR 12,6 [CI 95% 3,4-46,5]; p = 0,033) és trendszeru kapcsolatot a mortalitással (OR 3,6 [0,8-16,7]; p = 0,095). Emellett a nemzetközi normalizált ráta (INR) értéke (p = 0,001) és a szérumkreatinin-szint (p = 0,002) állt kapcsolatban a mortalitással. Az endoszkópos intervenció aránya szignifikáns összefüggésben volt a varix Paquet-stádiumával (p<0,001) és az ASA-pontszámmal (OR = 12,6 [3,4-46,5]; p = 0,033). Következtetés: Nyugat-Magyarországon magas az akut varixeredetu vérzés elofordulási gyakorisága. Az ASA-pontszám és a GBS jó prediktív faktor a betegségkimenetel és a transzfúziós igény vonatkozásában. A megfigyelt magas mortalitás és az endoszkópos ligatio alacsony aránya indokolja a kezelési stratégiák optimalizálását akut varixeredetu gastrointestinalis vérzés esetén. Orv Hetil. 2021; 162(31): 1252-1259. INTRODUCTION: Acute variceal gastrointestinal bleeding is associated with significant morbidity and mortality. OBJECTIVE: Our aim was to evaluate the characteristics and prognostic factors in the management of acute upper gastrointestinal bleeding in a large multi-center study from Hungary. METHOD: This prospective one-year study (between January 1, 2016 and December 31, 2016) involved six community hospitals in Western Hungary. Data collection included demographic characteristics, vital signs at admission, comorbidities, medications, time to hospital admission and endoscopy, laboratory results, endoscopic management, risk assessment using Glasgow-Blatchford Score (GBS), Rockall Score (RS) and the American Society of Anesthesiologists (ASA) Physical Status Score, transfusion requirements, length of hospital stay and mortality. RESULTS: 108 cases (male: 69.4%) of acute variceal gastrointestinal bleeding were registered during the 1-year period. Endoscopic therapeutic intervention was performed in 57.4%. On initial endoscopy, 39.8% of the patients were treated with sclerotherapy and 18.5% had ligation. 76.9% of the patients required blood transfusion. The overall mortality (including in-hospital bleedings) was 24.1%. The GBS predicted transfusions (AUC: 0.793; cut-off: GBS >8 points). The ASA Score was associated with transfusion (OR 7.6 [CI 95% 2.7-21.6]; p<0.001), endoscopic intervention (OR 12.6 [CI 95% 3.4-46.5]; p = 0.033), and showed similar trend with mortality (OR 3.6 [0.8-16.7]; p = 0.095). The increased international normalized ratio (INR) and creatinine levels were associated with mortality (p = 0.001 and p = 0.002). CONCLUSION: Incidence rates of acute variceal gastrointestinal bleeding in Western Hungary are high. The ASA Score, GBS predicted outcomes and transfusion requirements. The observed high mortality rates, coupled with relatively low rates of endoscopic ligation, warrant optimization of management strategies in acute variceal gastrointestinal bleeding. Orv Hetil. 2021; 162(31): 1252-1259.

Synthesis and In Vitro Characterization of Glycopeptide Drug Candidates Related to PACAP1-23.

The search for efficacious treatment of neurodegenerative and progressive neuroinflammatory diseases continues, as current therapies are unable to halt or reverse disease progression. PACAP represents one potential therapeutic that provides neuroprotection effects on neurons, and also modulates inflammatory responses and circulation within the brain. However, PACAP is a relatively long peptide hormone that is not trivial to synthesize. Based on previous observations that the shortened isoform PACAP1-23 is capable of inducing neuroprotection in vitro, we were inspired to synthesize shortened glycopeptide analogues of PACAP1-23. Herein, we report the synthesis and in vitro characterization of glycosylated PACAP1-23 analogues that interact strongly with the PAC1 and VPAC1 receptors, while showing reduced activity at the VPAC2 receptor.

The Equipment of the Tyrolean Iceman: Innovation versus Adaptation in the Cultural Evolution of Prehistoric Technologies.

This study argues that the equipment of the Tyrolean Iceman offers a unique perspective for understanding the macroevolutionary-scale functional continuity between later Pleistocene and Holocene human technologies. The Iceman was discovered in 1991 in the Ötztal Alps and can be dated to around 3300 BC, corresponding to the North Italian early Copper Age. In his gear there are several technologies which are rarely found at prehistoric archaeological sites, including archery equipment, a complete set of clothing, and a personal tool kit. Our paper will discuss this technological assemblage within the framework of cultural evolutionary theories (subsequently abbreviated as CET). According to the concept of cumulative culture, tools and technologies may become the subject of cultural "descent with modification" which leads to the emergence of complex technological innovations. In conventional narratives, the Neolithic represents the single greatest macroevolutionary transition in human cultural evolution, accompanied by a whole set of novel innovations and encompassing the transition to agriculture. However, as we highlight, the Iceman's equipment includes several technologies with a pre-Neolithic cultural origin. Earlier variants of these technologies were used by cultural groups belonging to the Mesolithic and even the Upper and Middle Paleolithic. Our main goal will be to present an explanatory framework for this macroevolutionary-scale technological continuity. In order to achieve this goal, we explore the heuristic value of two basic concepts of cultural evolutionary explanations-namely, the concepts of innovation and adaptation. Building on this background, we present an overview of the data currently available on the evolutionary history of each technological adaptation found in the equipment of the Iceman. Our results suggest that these technologies were not primarily cultural innovations, but simultaneously they were "obligatory" functional adaptations with a deep evolutionary history.

The Journey from Mental Disorder Towards Recovery.

No abstract available.

A modified culture medium and hyphae isolation method can increase quality of the RNA extracted from mycelia of a dimorphic fungal species.

The capability of RNA isolation with good efficiency and high quality is essential for a downstream application such as RNA sequencing. It requires successful cell culturing and an effective RNA isolation method. Although effective methods are available, production of the homogenous mycelia and extraction of good-quality mycelial RNA from true invasive hyphae, which penetrated into the agar plates, are difficult. To overcome these problems, the aim of this study was to develop technical modifications which allow production of homogenous mycelial biomass without extra stimuli agents and improve quality of the RNA extracted from the fungal hyphae. Our alternative culture medium was suitable for production both yeast-phase cells and hyphae of the Schizosaccharomyces japonicus and other dimorphic species, such as the Candida albicans, Saccharomyces cerevisiae, and Jaminaea angkorensis. To improve quality of the mycelial RNA, we developed an isolation procedure of the hyphal tip, which eliminated the unnecessary vacuoles-containing parts of the hyphae. To increase RNA quantity, we used glass beads in the RNA extraction protocol to achieve stronger breaking of the mycelial walls. All these modifications can also be useful for researchers working with other dimorphic fungi and can contribute to the higher comparability of the transcriptional data coming from yeast-phase cells and hyphae or even from different species.

The Delta-Specific Opioid Glycopeptide BBI-11008: CNS Penetration and Behavioral Analysis in a Preclinical Model of Levodopa-Induced Dyskinesia.

In previous work we evaluated an opioid glycopeptide with mixed µ/δ-opioid receptor agonism that was a congener of leu-enkephalin, MMP-2200. The glycopeptide analogue showed penetration of the blood-brain barrier (BBB) after systemic administration to rats, as well as profound central effects in models of Parkinson's disease (PD) and levodopa (L-DOPA)-induced dyskinesia (LID). In the present study, we tested the glycopeptide BBI-11008 with selective δ-opioid receptor agonism, an analogue of deltorphin, a peptide secreted from the skin of frogs (genus Phyllomedusa). We tested BBI-11008 for BBB-penetration after intraperitoneal (i.p.) injection and evaluated effects in LID rats. BBI-11008 (10 mg/kg) demonstrated good CNS-penetrance as shown by microdialysis and mass spectrometric analysis, with peak concentration levels of 150 pM in the striatum. While BBI-11008 at both 10 and 20 mg/kg produced no effect on levodopa-induced limb, axial and oral (LAO) abnormal involuntary movements (AIMs), it reduced the levodopa-induced locomotor AIMs by 50% after systemic injection. The N-methyl-D-aspartate receptor antagonist MK-801 reduced levodopa-induced LAO AIMs, but worsened PD symptoms in this model. Co-administration of MMP-2200 had been shown prior to block the MK-801-induced pro-Parkinsonian activity. Interestingly, BBI-11008 was not able to block the pro-Parkinsonian effect of MK-801 in the LID model, further indicating that a balance of mu- and delta-opioid agonism is required for this modulation. In summary, this study illustrates another example of meaningful BBB-penetration of a glycopeptide analogue of a peptide to achieve a central behavioral effect, providing additional evidence for the glycosylation technique as a method to harness therapeutic potential of peptides.

Highly-selective µ-opioid receptor antagonism does not block L-DOPA-induced dyskinesia in a rodent model.

OBJECTIVES: Dopamine-replacement utilizing L-DOPA is still the mainstay treatment for Parkinson's disease (PD), but often leads to development of L-DOPA-induced dyskinesia (LID), which can be as debilitating as the motor deficits. There is currently no satisfactory pharmacological adjunct therapy. The endogenous opioid peptides enkephalin and dynorphin are important co-transmitters in the direct and indirect striatofugal pathways and have been implicated in genesis and expression of LID. Opioid receptor antagonists and agonists with different selectivity profiles have been investigated for anti-dyskinetic potential in preclinical models. In this study we investigated effects of the highly-selective µ-opioid receptor antagonist CTAP (> 1200-fold selectivity for µ- over δ-opioid receptors) and a novel glycopeptide congener (gCTAP5) that was glycosylated to increase stability, in the standard rat LID model. RESULTS: Intraperitoneal administration (i.p.) of either 0.5 mg/kg or 1 mg/kg CTAP and gCTAP5 completely blocked morphine's antinociceptive effect (10 mg/kg; i.p.) in the warm water tail-flick test, showing in vivo activity in rats after systemic injection. Neither treatment with CTAP (10 mg/kg; i.p.), nor gCTAP5 (5 mg/kg; i.p.) had any effect on L-DOPA-induced limb, axial, orolingual, or locomotor abnormal involuntary movements. The data indicate that highly-selective µ-opioid receptor antagonism alone might not be sufficient to be anti-dyskinetic.

Behavioral pharmacology of the mixed-action delta-selective opioid receptor agonist BBI-11008: studies on acute, inflammatory and neuropathic pain, respiration, and drug self-administration.

RATIONALE AND OBJECTIVES: The present study characterized the behavioral pharmacology of a novel, mixed-action delta-selective (78:1) opioid receptor agonist, BBI-11008. This glycopeptide drug candidate was tested in assays assessing antinociception (acute, inflammatory, and neuropathic pain-like conditions) and side-effect endpoints (respiratory depression and drug self-administration). RESULTS: BBI-11008 had a 78-fold greater affinity for the delta opioid receptor than the mu receptor, and there was no binding to the kappa opioid receptor. BBI-11008 (3.2-100; 10-32 mg kg-1, i.v.) and morphine (1-10; 1-3.2 mg kg-1, i.v.) produced antinociceptive and anti-allodynic effects in assays of acute thermal nociception and complete Freund's adjuvant (CFA)-induced inflammatory pain, with BBI-11008 being less potent than morphine in both assays. BBI-11008 (1-18 mg kg-1, i.v.) had similar efficacy to gabapentin (10-56 mg kg-1, i.v.) in a spinal nerve ligation (SNL) model of neuropathic pain. In the respiration assay, with increasing %CO2 exposure, BBI-11008 produced an initial increase (32 mg kg-1, s.c.) and then decrease (56 mg kg-1, s.c.) in minute volume (MV) whereas morphine (3.2-32 mg kg-1, s.c.) produced dose-dependent decreases in MV. In the drug self-administration procedure, BBI-11008 did not maintain self-administration at any dose tested. CONCLUSIONS: These results suggest that the glycopeptide drug candidate possesses broad-spectrum antinociceptive and anti-allodynic activity across a range of pain-like conditions. Relative to morphine or fentanyl, the profile for BBI-11008 in the respiration and drug self-administration assays suggests that BBI-11008 may have less pronounced deleterious side effects. Continued assessment of this compound is warranted.

Biodegradability and Toxicity of Cellobiosides and Melibiosides.

In 2014, almost 16 million tons of surfactants were used globally for cleaning and industrial applications. As a result, massive quantities disperse into environmental compartments every day. There is great market interest in developing highly biodegradable, less-toxic, and renewable alternatives to currently used petroleum-based surfactants. Glycolipid surfactants, composed of a sugar head-group and lipid tail, are effective surfactants and emulsifiers with a high tolerance to electrolytes and are easily tailored to address specific needs. The green synthesis and surfactant characteristics of a suite of cellobiosides and melibiosides were recently described. The biodegradability and toxicity of 1°-alkyl-O-cellobiosides, 2°-alkyl-O-cellobiosides, and 1°-alkyl-O-melibiosides with straight-chain alkyl tails of 8, 10, and 12 are reported in this study. Biodegradability was assessed by quantifying mineralization (CO2 evolution). All of the glycosides were inherently biodegradable and most were readily biodegradable according to OECD and EPA definitions. The Microtox acute toxicity assay showed both chain length and head group had significant effects on toxicity, but most of the molecules were practically non-toxic according to EPA definitions with EC50 values > 100 mg L-1. Cytotoxicity to human lung (H1299) and keratinocyte cell lines (HaCaT) was measured by xCELLigence and MTS assays. Cytotoxicity values were comparable to similar glycosides previously reported. IC50 values were determined but, in general, exceeded surfactant concentrations that are found in the environment. These data demonstrate the promising nature of these molecules as green alternatives to petrochemical surfactants.