Evolution of Clinical Outcome Measures and Biomarkers in Sporadic Adult-Onset Degenerative Ataxia.

BACKGROUND: Sporadic adult-onset ataxias without known genetic or acquired cause are subdivided into multiple system atrophy of cerebellar type (MSA-C) and sporadic adult-onset ataxia of unknown etiology (SAOA). OBJECTIVES: To study the differential evolution of both conditions including plasma neurofilament light chain (NfL) levels and magnetic resonance imaging (MRI) markers. METHODS: SPORTAX is a prospective registry of sporadic ataxia patients with an onset >40 years. Scale for the Assessment and Rating of Ataxia was the primary outcome measure. In subgroups, blood samples were taken and MRIs performed. Plasma NfL was measured via a single molecule assay. Regional brain volumes were automatically measured. To assess signal changes, we defined the pons and middle cerebellar peduncle abnormality score (PMAS). Using mixed-effects models, we analyzed changes on a time scale starting with ataxia onset. RESULTS: Of 404 patients without genetic diagnosis, 130 met criteria of probable MSA-C at baseline and 26 during follow-up suggesting clinical conversion to MSA-C. The remaining 248 were classified as SAOA. At baseline, NfL, cerebellar white matter (CWM) and pons volume, and PMAS separated MSA-C from SAOA. NfL decreased in MSA-C and did not change in SAOA. CWM and pons volume decreased faster, whereas PMAS increased faster in MSA-C. In MSA-C, pons volume had highest sensitivity to change, and PMAS was a predictor of faster progression. Fulfillment of possible MSA criteria, NfL and PMAS were risk factors, CWM and pons volume protective factors for conversion to MSA-C. CONCLUSIONS: This study provides detailed information on differential evolution and prognostic relevance of biomarkers in MSA-C and SAOA. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Regional Brain and Spinal Cord Volume Loss in Spinocerebellar Ataxia Type 3.

BACKGROUND: Given that new therapeutic options for spinocerebellar ataxias are on the horizon, there is a need for markers that reflect disease-related alterations, in particular, in the preataxic stage, in which clinical scales are lacking sensitivity. OBJECTIVE: The objective of this study was to quantify regional brain volumes and upper cervical spinal cord areas in spinocerebellar ataxia type 3 in vivo across the entire time course of the disease. METHODS: We applied a brain segmentation approach that included a lobular subsegmentation of the cerebellum to magnetic resonance images of 210 ataxic and 48 preataxic spinocerebellar ataxia type 3 mutation carriers and 63 healthy controls. In addition, cervical cord cross-sectional areas were determined at 2 levels. RESULTS: The metrics of cervical spinal cord segments C3 and C2, medulla oblongata, pons, and pallidum, and the cerebellar anterior lobe were reduced in preataxic mutation carriers compared with controls. Those of cervical spinal cord segments C2 and C3, medulla oblongata, pons, midbrain, cerebellar lobules crus II and X, cerebellar white matter, and pallidum were reduced in ataxic compared with nonataxic carriers. Of all metrics studied, pontine volume showed the steepest decline across the disease course. It covaried with ataxia severity, CAG repeat length, and age. The multivariate model derived from this analysis explained 46.33% of the variance of pontine volume. CONCLUSION: Regional brain and spinal cord tissue loss in spinocerebellar ataxia type 3 starts before ataxia onset. Pontine volume appears to be the most promising imaging biomarker candidate for interventional trials that aim at slowing the progression of spinocerebellar ataxia type 3. © 2021 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

cis-Hydroxyproline-mediated pancreatic carcinoma growth inhibition in mice.

PURPOSE: This study addressed the question of whether the collagen metabolism modulator cis-4-Hydroxy-L-proline (CHP) is applicable for potential use as a therapeutic inhibitor of pancreatic carcinoma cell growth. METHODS: Cell proliferation, as well as quantification of apoptosis of murine Panc02 cells, was assessed after CHP treatment. Supplementary, the effect of CHP on tumor growth was examined in the subcutaneous Panc02 model in vivo. Mice received daily intraperitoneal injections of CHP (300, 400, and 500 mg/kg bw). In addition to the assessment of systemic parameters (blood count, enzyme activities), histology (HE) and immunohistochemistry (Ki-67) were performed from resected tumor specimens. RESULTS: Like reduction of metabolic activity, CHP also induced inhibition of cell growth in a dose-dependent manner, with however only slight increases in apoptosis. In vivo treatment of Panc02 tumors with CHP resulted in pronounced delay of tumor growth and decreases in tumor cell proliferation. Moreover, these effects were accompanied by a massive systemic leukocytosis as well increased leukocyte infiltration into the tumors subsequent to CHP therapy. CONCLUSIONS: CHP inhibits the proliferation of Panc02 tumor cells in a dose-dependent manner both in vitro and in vivo. Our presented data show that modulation of the collagen metabolism is an interesting strategy for treatment of pancreatic carcinoma.

Characterization of chemosensitivity and resistance of human cancer cell lines to platinum(II) versus platinum(IV) anticancer agents.

Platinum (Pt)(IV) complexes are thought to function as prodrugs for anticancer Pt(II) drugs. We studied two pairs of Pt(II)/Pt(IV) complexes to explore whether there were differences in their cytotoxic activities, their abilities to cause acquired resistance and their gene expression profiles in the resistant lines. Microtiter methods were used to evaluate the antiproliferative activity of cisplatin, oxoplatin, [trans-d,l-(1,2-diaminocyclo-hexane)]dichloroplatinum(II) [DACH-Pt(II)] and cis,trans-[trans-d,l-(1,2-diaminocyclo-hexane)]-dichlorodihydroxoplatinum(IV) [DACH-Pt(IV)] in a panel of 14 human cancer cell lines. Cisplatin and oxoplatin showed significant similar spectra of cytotoxicity, whereas DACH-Pt(II) and DACH-Pt(IV) did not. DACH-Pt(IV) required more than 24 h to reach full potency, whereas the other three Pt complexes achieved maximal activity in less than 24 h. The SISO cervical cell line was made four- to six-fold resistant to the four Pt complexes by weekly exposure to the respective agent. Glutathione (GSH) levels increased in all resistant lines except for the DACH-Pt(IV) resistant line. The catalytic concentrations of various redox enzymes (GSH transferase, GSH peroxidase, GSH reductase, catalase) were all unchanged in the resistant lines relative to the native line. Multidrug resistance protein 2 expression was detected in the cisplatin-resistant and oxoplatin-resistant cell lines but not in the native line. The transcription of 29,000 genes in the SISO lines resistant to either cisplatin or oxoplatin was studied by DNA-microarray methods and compared with the native line. Overall changes in gene transcription were very different between the cisplatin-resistant and oxoplatin-resistant cell lines. Thus, Pt(IV) complexes seem to have biological actions that distinguish them from their Pt(II) counterparts, even when they show cross-resistance.

Effervescent morphine results in faster relief of breakthrough pain in patients compared to immediate release morphine sulfate tablet.

Morphine tablets have been formulated to produce an easily ingested effervescent solution when placed in water. It was hypothesized that an aqueous solution would result in fast gastrointestinal transit with a more rapid onset of action compared to immediate release morphine sulfate (IRMS), which would be especially beneficial in treating breakthrough pain (BTP). In an open-label safety and efficacy study, effervescent morphine was given to 76 chronic cancer pain patients for treatment of BTP evaluating time until pain relief, global satisfaction and side effects. Results were compared to those obtained using an IRMS formulation in a preceding run-in period. For BTP, a mean dose of 28 mg of effervescent morphine (range 10-80 mg) resulted in a highly significant reduction of pain score (mean 7.8 to mean 3.2; P < 0.001). Efficacy was not different from that observed with IRMS. However, mean time until sufficient pain relief was significantly shorter than with IRMS (13 +/- 5.6 vs. 27 +/- 4.4 minutes; P < 0.01). The incidence of side effects was similar with the new morphine formulation and with IRMS. There was no relationship between the previous dose of the daily opioid and the effective dose of effervescent morphine. The dose for treatment of BTP was determined by individual titration and not predicted by the dose taken with the basic pain medication. Compared to IRMS, overall satisfaction for effervescent morphine was rated "superior" by 16.7%, and "better" by 63.2% of patients. Effervescent morphine offers an alternative for management of breakthrough cancer pain compared with the commonly used IRMS.

Cis-hydroxyproline-induced inhibition of pancreatic cancer cell growth is mediated by endoplasmic reticulum stress.

AIM: To investigate the biological effects of cis-hydroxyproline (CHP) on the rat pancreatic carcinoma cell line DSL6A, and to examine the underlying molecular mechanisms. METHODS: The effect of CHP on DSL6A cell proliferation was assessed by using BrdU incorporation. The expression of focal adhesion kinase (FAK) was characterized by Western blotting and immunofluorescence. Induction of endoplasmic reticulum (ER) stress was investigated by using RT-PCR and Western blotting for the glucose-related protein-78 (GRP78) and growth arrest and DNA inducible gene (GADD153). Cell viability was determined through measuring the metabolic activity based on the reduction potential of DSL6A cells. Apoptosis was analyzed by detection of caspase-3 activation and cleavage of poly(ADP-ribose) polymerase (PARP) as well as DNA laddering. RESULTS: In addition to inhibition of proliferation, incubation with CHP induced proteolytic cleavage of FAK and a delocalisation of the enzyme from focal adhesions, followed by a loss of cell adherence. Simultaneously, we could show an increased expression of GRP78 and GADD153, indicating a CHP-mediated activation of the ER stress cascade in the DSL6A cell line. Prolonged incubation of DSL6A cells with CHP finally resulted in apoptotic cell death. Beside L-proline, the inhibition of intracellular proteolysis by addition of a broad spectrum protease inhibitor could abolish the effects of CHP on cellular functions and the molecular processes. In contrast, impeding the activity of apoptosis-executing caspases had no influence on CHP-mediated cell damage. CONCLUSION: Our data suggest that the initiation of ER stress machinery by CHP leads to an activation of intracellular proteolytic processes, including caspase-independent FAK degradation, resulting in damaging pancreatic carcinoma cells.

Antimicrobial efficacy of amine fluoride based tooth gels compared to a toothpaste in a phase 2/step 2 in-vitro test model.

INTRODUCTION: The aim of the present study was to determine the antimicrobial effect of various gel formulations on plaque formation; different tooth gels were compared to a toothpaste containing comparable antimicrobial ingredients with regard to its microbiocidal activity. The study was conducted under the assumption, that a chief requirement for the prevention of plaque formation is the combination of mechanical removal and antimicrobial activity, and not the sole capability of mechanical plaque removal. METHODS: Ledermix(®) fluoride gel as commercially available with preservative, and without preservative and perfume oils, Elmex(®) gelée, and Meridol(®) toothpaste were tested in a standardized in-vitro test modification of the quantitative suspension test EN 1040. Instead of testing in a suspension, the respective product was directly placed on a standardized contaminated sterile stainless steel disk without adding any bio-burden. 50% egg yolk in Aqua dest. was used as a neutralizer. RESULTS: Within 1 min, Elmex(®) gelée showed a RF >5 log(10) against S. pyogenes and S. sanguinis. Against S. mutans, a log(10) RF of ≥5 was achieved after 2 min, against C. albicans after 5 min, and against P. aeruginosa after 10 min S. aureus was the most difficult organisms to be reduced. After an application time of 10 min, only a log(10) RF of 2.4 was achieved. Ledermix exceeded the antimicrobial efficacy of Elmex(®) gelée against S. mutans and C. albicans and was already effective against these organisms after 1 min, but did not show the same antimicrobial efficacy as Elmex(®) gelée against P. aeruginosa. Similar to Elmex(®) gelée, a required reduction of >5 log(10) for antimicrobials under no organic challenge was not achieved against S. aureus. Ledermix(®) fluoride gel without preservatives and Ledermix(®) fluoride gel without preservatives and perfume oil did not show the antimicrobial efficacy of the standard Ledermix(®) fluoride gel formulation, indicating that the observed antimicrobial efficacy is chiefly based on the preservative, and possibly the perfume oil. Compared to the tested gels, Meridol(®) toothpaste was less effective and reached any antimicrobial effect >5 log(10) only against S. sanguinis after 10 min. CONCLUSION: All unmodified tested gels showed an antimicrobial effect. Because no relevant antimicrobial efficacy against plaque forming bacteria was achieved within 2 min, in practice, an anti-plaque forming effect based on the antimicrobial action of gels cannot be assumed when used in the oral cavity. However, the results of the present study indicate that the antimicrobial efficacy of gels is determined by their formulation and that for the prevention of plaque formation the combination of mechanical removal and antimicrobial activity is not the chief requirement only, but a sustained antimicrobial effect may be of greater importance.