Enriched stable 204Pb as tracer at ultra-low levels in clinical investigations.

The potential of enriched Pb (204Pb) was assessed to monitor pathways of trace levels of Pb in the pg range within the human body via isotope pattern variation in situations where natural lead cannot be used as a tracer due to regulatory limitations. Isotope ratio measurements were accomplished by means of (multi-collector) inductively coupled plasma mass spectrometry including a comparison of single and multi-collector ICP-MS for low-level 204Pb assessment. Isotopic pattern results from a blend of a large quantity of the element with a natural isotopic composition and an enriched stable isotope at orders of magnitude lower levels pose a nontrivial analytical problem. Isotope pattern deconvolution was successfully applied as mathematical tool based on multiple linear regressions. The method allowed for deconvolving the isotope pattern from measured isotope ratios without knowing the quantities of different isotope sources incorporated and mixed into the sample at levels of < 1 pg 204Pb/g blood. The objective of this manuscript is to evaluate and summarize the analytical aspects for Pb isotope pattern deconvolution based on the results of a clinical trial, where a 204Pb-enriched isotope tracer was applied to investigate the bioavailability of orally applied Pb along with purified clinoptilolite tuff as potential supplement. This unique approach allows to reduce tracer amounts to harmless levels to human health, which are in accordance with the legal regulative to study enrichment levels of < 0.01% in human blood.

Safety and efficacy of purified clinoptilolite-tuff treatment in patients with irritable bowel syndrome with diarrhea: Randomized controlled trial.

BACKGROUND: Irritable bowel syndrome (IBS) is a highly prevalent gastrointestinal disorder with poor response to treatment. IBS with predominant diarrhea (IBS-D) is accompanied by abdominal pain as well as high stool frequency and urgency. Purified clinoptilolite-tuff (PCT), which is approved by the Food and Drug Administration for use as a dietary supplement with the brand name G-PUR®, has previously shown therapeutic potential in other indications based on its physical adsorption capacity. AIM: To assess whether symptoms of IBS-D can be ameliorated by oral treatment with PCT. METHODS: In this randomized, placebo-controlled, double-blind pilot study, 30 patients with IBS-D diagnosis based on Rome IV criteria were enrolled. Following a 4-wk run-in phase, 14 patients were randomized to receive a 12-wk treatment with G-PUR® (2 g three times daily), and 16 patients received placebo. The relief from IBS-D symptoms as measured by the proportion of responders according to the Subject's Global Assessment (SGA) of Relief was assessed as the primary outcome. For the secondary outcomes, validated IBS-D associated symptom questionnaires, exploratory biomarkers and microbiome data were collected. RESULTS: The proportions of SGA of Relief responders after 12 wk were comparable in both groups, namely 21% in the G-PUR® group and 25% in the placebo group. After 4 wk of treatment, 36% of patients in the G-PUR® group vs 0% in the placebo group reported complete or considerable relief. An improvement in daily abdominal pain was noted in 94% vs 83% (P = 0.0353), and the median number of days with diarrhea per week decreased by 2.4 d vs 0.3 d in the G-PUR® and placebo groups, respectively. Positive trends were observed for 50% of responders in the Bristol Stool Form Scale. Positive trends were also noted for combined abdominal pain and stool consistency response and the Perceived Stress Questionnaire score. Only 64% in the G-PUR® group compared to 86% in the placebo group required rescue medication intake during the study. Stool microbiome studies showed a minor increase in diversity in the G-PUR® group but not in the placebo group. No PCT-related serious adverse events were reported. CONCLUSION: In this randomized, double-blind, placebo-controlled study, the PCT product, G-PUR®, demonstrated safety and clinical benefit towards some symptoms of IBS-D, representing a promising novel treatment option for these patients.

Purified Clinoptilolite-Tuff as an Efficient Sorbent for Gluten Derived from Food.

Various gluten-related diseases (celiac disease, wheat allergy, gluten sensitivity) are known and their incidence is growing. Gluten is a specific type of plant storage protein that can impair the health of gluten-prone persons following consumption, depending on the origin. The most severe effects are induced by wheat, barley, and rye. The only treatment is based on the absolute avoidance of those foods, as even traces might have severe effects on human well-being. With the goal of binding gluten impurities after ingestion, an in vitro setting was created. A special processed kind of zeolite, purified clinoptilolite-tuff (PCT), was implemented as an adsorber of gluten derived from different origins. Zeolites are known for their excellent sorption capacities and their applications in humans and animals have been studied for a long time. Tests were also performed in artificial gastric and intestinal fluids, and the adsorption capacity was determined via a certified validated method (ELISA). Depending on the kind of gluten source, 80-130 µg/mg of gluten were bound onto PCT. Hence, purified clinoptilolite-tuff, which was successfully tested for wheat, barley, and rye, proved to be suitable for the adsorption of gluten originating from different kinds of crops. This result might form the basis for an expedient human study in the future.

Topically administered purified clinoptilolite-tuff for the treatment of cutaneous wounds: A prospective, randomised phase I clinical trial.

In an ageing society, chronic ulcers pose an increasingly relevant healthcare issue associated with significant morbidity and an increasing financial burden. Hence, there is an unmet medical need for novel, cost-effective therapies that improve healing of chronic cutaneous wounds. This prospective, randomised, open-label, phase I trial investigated the safety and tolerability of topically administered purified clinoptilolite-tuff (PCT), mainly consisting of the naturally occurring zeolite-mineral clinoptilolite, in artificial wounds in healthy male volunteers compared to the standard of care (SoC). We found that topically administered PCT was safe for therapeutic application in acute wounds in healthy male volunteers. No significant differences in wound healing or wound conditions were observed compared to SoC-treated wounds. However, we found a significantly higher proportion of CD68-positive cells and a significantly lower proportion of α-smooth muscle actin-positive cells in PCT-treated wounds. Scanning electron microscopy revealed PCT particles in the restored dermis in some cases. However, these did not impede wound healing or clinical symptoms. Hence, purified PCT could represent an attractive, cost-effective wound treatment promoting the process of healing.

Concomitant oral intake of purified clinoptilolite tuff (G-PUR) reduces enteral lead uptake in healthy humans.

Lead exposure can cause substantial organ damage. Enteral lead absorption may be reduced by concomitant intake of clinoptilolite tuff, a zeolite from natural sources. This study aimed to assess the effect of purified clinoptilolite tuff (G-PUR) on enteral lead uptake in adults using stable lead isotope 204Pb as a tracer. In this randomized, placebo-controlled, double-blind, parallel-group study, 42 healthy participants were randomized to receive oral G-PUR 2.0 g, 2 * 2.0 g, or placebo, together with 2.5 µg of 204Pb in water. The enrichment of 204Pb caused by the tracer in blood and urine was measured by mass spectrometry. G-PUR was well tolerated. The mean maximum 204Pb enrichment of 0.505% of total blood lead was significantly higher (p < 0.0001) in the placebo group compared to G-PUR 2.0 g (0.073%) or G-PUR 2 * 2.0 g (0.057%) group. Normalized 204Pb AUC0-192 was 86.5, 11.9, and 8.5% * h without and with G-PUR 2.0 g, and G-PUR 2 * 2.0 g, respectively (p < 0.0001 vs. placebo). This smaller 204Pb exposure was paralleled by a reduced urinary excretion in subjects receiving G-PUR. Concomitant oral intake of purified clinoptilolite tuff reduced enteral uptake of 204Pb in healthy humans by approximately 90%. The reduced bioavailability is demonstrable by a decrease of 204Pb tracer enrichment in blood and urine.Trial registration: clinicaltrials.gov identifier: NCT04138693, registered 24/10/2019.

Binding and neutralization of C. difficile toxins A and B by purified clinoptilolite-tuff.

Clostridioides difficile (C. difficile) infection is a major public health problem worldwide. The current treatment of C. difficile-associated diarrhea relies on the use of antibacterial agents. However, recurrences are frequent. The main virulence factors of C. difficile are two secreted cytotoxic proteins toxin A and toxin B. Alternative research exploring toxin binding by resins found a reduced rate of recurrence by administration of tolevamer. Hence, binding of exotoxins may be useful in preventing a relapse provided that the adsorbent is innocuous. Here, we examined the toxin binding capacity of G-PUR®, a purified version of natural clinoptilolite-tuff. Our observations showed that the purified clinoptilolite-tuff adsorbed clinically relevant amounts of C. difficile toxins A and B in vitro and neutralized their action in a Caco-2 intestinal model. This conclusion is based on four independent sets of findings: G-PUR® abrogated toxin-induced (i) RAC1 glucosylation, (ii) redistribution of occludin, (iii) rarefaction of the brush border as visualized by scanning electron microscopy and (iv) breakdown of the epithelial barrier recorded by transepithelial electrical resistance monitoring. Finally, we confirmed that the epithelial monolayer tolerated G-PUR® over a wide range of particle densities. Our findings justify the further exploration of purified clinoptilolite-tuff as a safe agent in the treatment and/or prevention of C. difficile-associated diarrhea.

Clinoptilolite in Dextran Sulphate Sodium-Induced Murine Colitis: Efficacy and Safety of a Microparticulate Preparation.

Background: Clinoptilolite is an aluminium silicate of natural origin; the microporous structure and the net negative charge of its crystal lattice allows for adsorption of ions, toxins, inflammatory mediators, and some microorganisms. We generated 2 preparations of purified clinoptilolite, which differed by about 10-fold in particle size, ie, a standard powder (GHC1) and a microparticulate fraction (GHC2) with a size of 3.6 µm and 0.39 µm (d50) respectively. These were examined for their ability to accelerate the recovery of mice from DSS (dextran sulphate sodium)-induced intestinal inflammation. Methods: Efficacy of clinoptilolite preparations was investigated by administering DSS-treated mice twice daily with 30 mg GHC2 or GHC1 for 5 consecutive days, followed by 5 days of recovery without DSS. To explore the safety of the microparticulate preparation (GHC2), mice were subjected to 4 cycles of DSS-exposure. We specifically verified that clinoptilolite microparticles were not systemically bioavailable by examining the gut tissue and the liver for the accumulation of microparticles by transmission electron microscopy. Results: Treatment of mice with GHC2 was superior to GHC1 and as effective as the reference compound 5-aminosalicylic acid in ameliorating the damage induced by the exposure to DSS. In addition, no clinoptilolite particle was observed in the intestinal epithelial layer, gut-associated lymph follicles, or in the liver. Conclusion: Our observations confirm that a microparticulate preparation of clinoptilolite is safe and effective in a murine model of inflammatory bowel disease and supports the hypothesis that the adsorptive capacity of clinoptilolite is of potential therapeutic relevance.

Preferential cataclastic grain size reduction of feldspar in deformation bands in poorly consolidated arkosic sands.

This study presents microstructural as well as bulk and mineral chemical investigations of deformation bands in uncemented, friable arkosic sands of Miocene age (Vienna Basin, Austria). Our microstructural study indicates grain size reduction by grain flaking in deformation bands with small offsets (0.5-8 cm), and dominant intragranular fracturing and cataclasis of altered feldspar grains at larger displacements (up to 60 cm). Relative to quartz, the sericitized feldspar grains are preferably fractured and abraded, which additionally leads to an enrichment of mainly phyllosilicates by mechanical expulsion from feldspar. Both cataclasis of quartz and feldspar grains and enrichment of phyllosilicates result in grain size reduction within the deformation bands. The measured reduction in porosity of up to 20% is in some cases associated with a permeability reduction, reflected in the retention of iron-oxide rich fluids along deformation bands. These deformation bands formed at very shallow burial depths in unconsolidated sediments indicate that fault sealing may occur in the absence of chemical alteration of the deformation bands and lead to a compartmentalization of a groundwater or hydrocarbon reservoir.

Magmatism at the Eurasian-North American modern plate boundary: Constraints from alkaline volcanism in the Chersky Belt (Yakutia).

The Chersky seismic belt (NE-Russia) forms the modern plate boundary of the Eurasian-North American continental plate. The geodynamic evolution of this continent-continent setting is highly complex and remains a matter of debate, as the extent and influence of the Mid-Arctic Ocean spreading center on the North Asian continent since the Eocene remains unclear. The progression from a tensional stress regime to a modern day transpressional one in the Chersky seismic belt, makes the understanding even more complicated. The alkaline volcanism that has erupted along the Chersky range from Eocene through to the Recent can provide constraints on the geodynamic evolution of this continental boundary, however, the source and petrogenetic evolution of these volcanic rocks and their initiating mechanisms are poorly understood. We studied basanites of the central Chersky belt, which are thought to represent the first alkaline volcanic activity in the area, after initial opening of the Arctic Ocean basin. We present mineral and bulk rock geochemical data as well as Sr-Nd-Pb-Hf isotopes of the alkaline suite of rocks combined with new precise K-Ar and 40Ar/39Ar dating, and discuss an integrated tectono-magmatic model for the Chersky belt. Our findings show that the basanites were generated from a homogeneous asthenospheric mantle reservoir with an EM-1 isotopic flavor, under relatively 'dry' conditions at segregation depths around 110 km and temperatures of ~ 1500 °C. Trace element and isotope systematics combined with mantle potential temperature estimates offer no confirmation of magmatism related to subduction or plume activity. Mineral geochemical and petrographical observations together with bulk geochemical evidence indicate a rapid ascent of melts and high cooling rates after emplacement in the continental crust. Our preferred model is that volcanism was triggered by extension and thinning of the lithosphere combined with adiabatic upwelling of the underlying mantle at 37 Ma. This suggests that at that time, rift tectonics in the Mid-Arctic Ocean most likely had also affected the North-Asian continent, causing volcanic activity in the Chersky belt, before the regional geodynamic regime changed from a tensional to compressional. Our conclusions contribute not only to the understanding of volcanism in the Chersky seismic belt (NE-Russia) but also to general aspects of plate dynamics between the Eurasian and North American continent.

Displacement-length scaling of brittle faults in ductile shear.

Within a low-grade ductile shear zone, we investigated exceptionally well exposed brittle faults, which accumulated antithetic slip and rotated into the shearing direction. The foliation planes of the mylonitic host rock intersect the faults approximately at their centre and exhibit ductile reverse drag. Three types of brittle faults can be distinguished: (i) Faults developing on pre-existing K-feldspar/mica veins that are oblique to the shear direction. These faults have triclinic flanking structures. (ii) Wing cracks opening as mode I fractures at the tips of the triclinic flanking structures, perpendicular to the shear direction. These cracks are reactivated as faults with antithetic shear, extend from the parent K-feldspar/mica veins and form a complex linked flanking structure system. (iii) Joints forming perpendicular to the shearing direction are deformed to form monoclinic flanking structures. Triclinic and monoclinic flanking structures record elliptical displacement-distance profiles with steep displacement gradients at the fault tips by ductile flow in the host rocks, resulting in reverse drag of the foliation planes. These structures record one of the greatest maximum displacement/length ratios reported from natural fault structures. These exceptionally high ratios can be explained by localized antithetic displacement along brittle slip surfaces, which did not propagate during their rotation during surrounding ductile flow.

Polybaric petrogenesis of Neogene alkaline magmas in an extensional tectonic environment: Viliga Volcanic Field, northeast Russia.

Neogene alkaline intraplate volcanic rocks from the Viliga Volcanic Field (NE-Russia) were studied both to precisely characterize their geochemical composition and to unravel their petrogenetic history. The sampled volcanic rocks crop out within the voluminous calc-alkaline sequences of the Cretaceous Okhostk-Chukokta Volcanic Belt, an Andean-type arc formed during subduction of the Paleo-Pacific Plate beneath modern far-east Asia. The mantle xenolith- and xenocryst-bearing basanites and nephelinites have intraplate ocean island basalt-type geochemical features. Sr and Nd isotopes combined with major and trace element systematics and rare-earth element modeling suggest polybaric melt generation of these alkaline magmatic rocks from a homogeneous garnet peridotite facies source with minor involvement of spinel peridotite facies partial melts. The basanite samples indicate segregation PT conditions around 1500 °C at 33-38 kbar whilst the nephelinites reflect smaller melt fractions segregated at over 40 kbar. During ascent, olivine (up to 7%) was the main fractionating phase in the basanites; whereas in the nephelinites, both olivine and minor clinopyroxene fractionation occurred. Crustal contamination during ascent was insignificant. We argue that the melt generation of these alkaline magmas from the Viliga Volcanic Field was triggered by an extending lithosphere resulting in upwelling asthenosphere and decompression melting, analogous to geodynamic models of the coeval alkaline volcanic rocks along the adjacent North Pacific continental margins, rather than by subduction- or plume-related processes.