Quantification of Solid Embryonic Cerebellar Graft Volume in a Degenerative Ataxia Model.

Substitution of lost neurons by neurotransplantation would be a possible management of advanced degenerative cerebellar ataxias in which insufficient cerebellar reserve remains. In this study, we examined the volume and structure of solid embryonic cerebellar grafts in adult Lurcher mice, a model of olivocerebellar degeneration, and their healthy littermates. Grafts taken from enhanced green fluorescent protein (EGFP)-positive embryos were injected into the cerebellum of host mice. Two or six months later, the brains were examined histologically. The grafts were identified according to the EGFP fluorescence in frozen sections and their volumes were estimated using the Cavalieri principle. For gross histological evaluation, graft-containing slices were processed using Nissl and hematoxylin-eosin staining. Adjustment of the volume estimation approach suggested that it is reasonable to use all sections without sampling, but that calculation of values for up to 20% of lost section using linear interpolation does not constitute substantial error. Mean graft volume was smaller in Lurchers than in healthy mice when examined 6 months after the transplantation. We observed almost no signs of graft destruction. In some cases, compact grafts disorganized the structure of the host's cerebellar cortex. In Lurchers, the grafts had a limited contact with the host's cerebellum. Also, graft size was of greater variability in Lurchers than in healthy mice. The results are in compliance with our previous findings that Lurcher phenotype-associated factors have a negative effect on graft development. These factors can hypothetically include cerebellar morphology, local tissue milieu, or systemic factors such as immune system abnormalities.

Lipid Nanoparticles Deliver mRNA to the Brain after an Intracerebral Injection.

Neurological disorders are often debilitating conditions with no cure. The majority of current therapies are palliative rather than disease-modifying; therefore, new strategies for treating neurological disorders are greatly needed. mRNA-based therapeutics have great potential for treating such neurological disorders; however, challenges with delivery have limited their clinical potential. Lipid nanoparticles (LNPs) are a promising delivery vector for the brain, given their safer toxicity profile and higher efficacy. Despite this, very little is known about LNP-mediated delivery of mRNA into the brain. Here, we employ MC3-based LNPs and successfully deliver Cre mRNA and Cas9 mRNA/Ai9 sgRNA to the adult Ai9 mouse brain; greater than half of the entire striatum and hippocampus was found to be penetrated along the rostro-caudal axis by direct intracerebral injections of MC3 LNP mRNAs. MC3 LNP Cre mRNA successfully transfected cells in the striatum (∼52% efficiency) and hippocampus (∼49% efficiency). In addition, we demonstrate that MC3 LNP Cas9 mRNA/Ai9 sgRNA edited cells in the striatum (∼7% efficiency) and hippocampus (∼3% efficiency). Further analysis demonstrates that MC3 LNPs mediate mRNA delivery to multiple cell types including neurons, astrocytes, and microglia in the brain. Overall, LNP-based mRNA delivery is effective in brain tissue and shows great promise for treating complex neurological disorders.

Experimental Treatment with Edaravone in a Mouse Model of Spinocerebellar Ataxia 1.

Edaravone is a mitochondrially targeted drug with a suggested capability to modify the course of diverse neurological diseases. Nevertheless, edaravone has not been tested yet in the context of spinocerebellar ataxia 1 (SCA1), an incurable neurodegenerative disease characterized mainly by cerebellar disorder, with a strong contribution of inflammation and mitochondrial dysfunction. This study aimed to address this gap, exploring the potential of edaravone to slow down SCA1 progression in a mouse knock-in SCA1 model. SCA1154Q/2Q and healthy SCA12Q/2Q mice were administered either edaravone or saline daily for more than 13 weeks. The functional impairments were assessed via a wide spectrum of behavioral assays reflecting motor and cognitive deficits and behavioral abnormalities. Moreover, we used high-resolution respirometry to explore mitochondrial function, and immunohistochemical and biochemical tools to assess the magnitude of neurodegeneration, inflammation, and neuroplasticity. Data were analyzed using (hierarchical) Bayesian regression models, combined with the methods of multivariate statistics. Our analysis pointed out various previously documented neurological and behavioral deficits of SCA1 mice. However, we did not detect any plausible therapeutic effect of edaravone on either behavioral dysfunctions or other disease hallmarks in SCA1 mice. Thus, our results did not provide support for the therapeutic potential of edaravone in SCA1.

[CME-Sonography 104: Angiomyolipomas]. / CME-Sonografie 104: Angiomyolipome.

CME-Sonography 104: Angiomyolipomas Abstract. Angiomyolipomas are the most common benign kidney tumors. Approximately 80 % are spontaneously occurring tumors, the majority <1.0 cm (approximately 54 %). These tumors do not grow and are harmless. Tumors between 1.0 and 2.5 cm (approx. 44 %) also very rarely grow and remain harmless. Larger tumors are rarely found (about 2 %). These must be carefully monitored, as tumors >4.0 cm can spontaneously manifest dangerous bleeding. Apart from spontaneous occurrence, there are also angiomyolipomas in the context of the tuberous sclerosis complex (TCS). Such tumors are often larger and combined with renal cysts and renal cell carcinomas. Measuring the of echo intensity is important for diagnosis.

Consensus Paper: Strengths and Weaknesses of Animal Models of Spinocerebellar Ataxias and Their Clinical Implications.

Spinocerebellar ataxias (SCAs) represent a large group of hereditary degenerative diseases of the nervous system, in particular the cerebellum, and other systems that manifest with a variety of progressive motor, cognitive, and behavioral deficits with the leading symptom of cerebellar ataxia. SCAs often lead to severe impairments of the patient's functioning, quality of life, and life expectancy. For SCAs, there are no proven effective pharmacotherapies that improve the symptoms or substantially delay disease progress, i.e., disease-modifying therapies. To study SCA pathogenesis and potential therapies, animal models have been widely used and are an essential part of pre-clinical research. They mainly include mice, but also other vertebrates and invertebrates. Each animal model has its strengths and weaknesses arising from model animal species, type of genetic manipulation, and similarity to human diseases. The types of murine and non-murine models of SCAs, their contribution to the investigation of SCA pathogenesis, pathological phenotype, and therapeutic approaches including their advantages and disadvantages are reviewed in this paper. There is a consensus among the panel of experts that (1) animal models represent valuable tools to improve our understanding of SCAs and discover and assess novel therapies for this group of neurological disorders characterized by diverse mechanisms and differential degenerative progressions, (2) thorough phenotypic assessment of individual animal models is required for studies addressing therapeutic approaches, (3) comparative studies are needed to bring pre-clinical research closer to clinical trials, and (4) mouse models complement cellular and invertebrate models which remain limited in terms of clinical translation for complex neurological disorders such as SCAs.

The Coiled-Coil Forming Peptide (KVSALKE)5 Is a Cell Penetrating Peptide that Enhances the Intracellular Delivery of Proteins.

Protein-based therapeutics have the potential to treat a variety of diseases, however, safe and effective methods for delivering them into cells need to be developed before their clinical potential can be realized. Peptide fusions have great potential for improving intracellular delivery of proteins. However, very few peptides have been identified that can increase the intracellular delivery of proteins, and new peptides that can enhance intracellular protein delivery are greatly needed. In this report, the authors demonstrate that the coiled-coil forming peptide (KVSALKE)5 (termed K5) can function as a cell penetrating peptide (CPP), and can also complex other proteins that contain its partner peptide E5. It is shown here that GFP and Cas9 fused to the K5 peptide has dramatically enhanced cell uptake in a variety of cell lines, and is able to edit neurons and astrocytes in the striatum and hippocampus of mice after a direct intracranial injection. Collectively, these studies demonstrate that the coiled-coil forming peptide (KVSALKE)5 is a new class of multifunctional CPPs that has great potential for improving the delivery of proteins into cells and in vivo.

[CME-Sonography 102: Acute Epigastric Pain]. / CME-Sonografie 102: Akute Oberbauchschmerzen.

CME-Sonography 102: Acute Epigastric Pain Abstract. Acute upper abdominal pain is a common issue in emergency medicine. Next to the anamnesis and laboratory findings, sonography plays a leading role in differentiation. In addition to the direct representation of the respective pathologies (gallstones, duct dilatations, tumors), the clinical-sonographic examination, the so-called palpation under ultrasound view, is also important. In this article we aim to show the way of diagnosing acute epigastric pain.

[CME-Sonography 102/Answers: Acute Epigastric Pain]. / CME-Sonografie 102/Antworten: Akute Oberbauchschmerzen.

CME-Sonography 102/Answers: Acute Epigastric Pain Abstract. Acute upper abdominal pain is a common issue in emergency medicine. Next to the anamnesis and laboratory findings, sonography plays a leading role in differentiation. In addition to the direct representation of the respective pathologies (gallstones, duct dilatations, tumors), the clinical-sonographic examination, the so-called palpation under ultrasound view, is also important. In this article we aim to show the way of diagnosing acute epigastric pain.

[CME Sonography 99/Answers: Kidney Echo Changes]. / CME-Sonografie 99/Antworten: Veränderungen der Nierenechogenität.

CME Sonography 99/Answers: Kidney Echo Changes Abstract. Normal kidneys have a smooth contour, kidney length 9-12 cm, volume 90-180 ml/1.73 m2 body surface, parenchyma width 13-18 mm and an echogenicity of the cortex which is hypoechoic compared to the liver and spleen, as well as medullary pyramids, which are again hypoechoic compared to the cortex. Digital ultrasound images enable the echogenicity to be measured between 0 (black) and 255 (white). A normal quotient between the echogenicity of the liver and the renal cortex is 1.0-1.4. A normal quotient between the spleen and the renal cortex is> 1.0. The normal quotient between the renal cortex and the renal medulla is> 1.0. In diffuse renal parenchyma diseases, changes in echogenicity are observed. There are kidneys with increased echogenicity of the cortex and a ratio to liver and spleen of <1.0 and kidneys with hyperechoic renal medulla and the ratio between renal cortex and renal medulla <1.0.

[CME Sonography 98/Answers: Chest Ultrasound]. / CME-Sonografie 98/Antworten: Thoraxsonografie.

CME Sonography 98/Answers: Chest Ultrasound Abstract. Chest sonography has long been an important part of ultrasound diagnostics. Historically, the first evidence found in chest sonography were pleural effusions. Peripheral consolidations (pneumonia, tumors, pulmonary embolism) and pneumothorax were added later. The COVID-19 pandemic with often massive lung infestation has significantly increased the interest in thoracic sonography. The partially specific changes caused by COVID-19 are presented in this article.

[CME Sonography 99: Kidney Echo Changes]. / CME-Sonografie 99: Veränderungen der Nierenechogenität.

CME Sonography 99: Kidney Echo Changes Abstract. Normal kidneys have a smooth contour, kidney length 9-12 cm, volume 90-180 ml/1.73 m2 body surface, parenchyma width 13-18 mm and an echogenicity of the cortex which is hypoechoic compared to the liver and spleen, as well as medullary pyramids, which are again hypoechoic compared to the cortex. Digital ultrasound images enable the echogenicity to be measured between 0 (black) and 255 (white). A normal quotient between the echogenicity of the liver and the renal cortex is 1.0-1.4. A normal quotient between the spleen and the renal cortex is >1.0. The normal quotient between the renal cortex and the renal medulla is >1.0. In diffuse renal parenchyma diseases, changes in echogenicity are observed. There are kidneys with increased echogenicity of the cortex and a ratio to liver and spleen of <1.0 and kidneys with hyperechoic renal medulla and the ratio between renal cortex and renal medulla <1.0.

[CME Sonography 98: Chest Ultrasound]. / CME-Sonografie 98: Thoraxsonografie.

CME Sonography 98: Chest Ultrasound Abstract. Chest sonography has long been an important part of ultrasound diagnostics. Historically, the first evidence found in chest sonography were pleural effusions. Peripheral consolidations (pneumonia, tumors, pulmonary embolism) and pneumothorax were added later. The COVID-19 pandemic with often massive lung infestation has significantly increased the interest in thoracic sonography. The partially specific changes caused by COVID-19 are presented in this article.

[Detection of kidney stones - Imaging and laboratory chemistry including urine analysis]. / Nachweis von Nierensteinen.

Detection of kidney stones - Imaging and laboratory chemistry including urine analysis Abstract. Nephrolithiasis is often the first manifestation of renal colic. This is usually caused by ureteral stones, but also by blood clots, lost papillary necrosis, tumor-related ureteral stenosis or urogenital TB. Today, kidney stones are primarily detected by ultrasound. Although the CT examination without contrast agent is also highly sensitive, because of the radiation exposure, it is only recommended if the findings are unclear. In addition to a careful anamnesis, supplementary diagnostics with urine tests (hematuria and crystalluria, possibly 24-hour urine collection) and serological determinations (creatinine, sodium, potassium, calcium, uric acid, inorganic phosphorus) are important.

[CME Sonography 96/Answers: Spleen Changes]. / CME-Sonografie 96/Antworten: Milzveränderungen.

CME Sonography 96/Answers: Spleen Changes Abstract. The spleen is a somewhat neglected organ but there are some changes that must not be neglected. Splenomegaly, caused worldwide most commonly by malaria, can occur in other infections, in portal vein thrombosis or in portal hypertension in the context of liver cirrhosis. An accessory spleen is also often found. Sometimes after splenectomy, small remains are found which may later hypertrophy. Where focal changes are concerned, we differentiate between various forms of spleen cysts, lymphomas, metastases, and benign tumors.

[CME Sonography 96: Spleen Changes]. / CME-Sonografie 96: Milzveränderungen.

CME Sonography 96: Spleen Changes Abstract. The spleen is a somewhat neglected organ but there are some changes that must not be neglected. Splenomegaly, caused worldwide most commonly by malaria, can occur in other infections, in portal vein thrombosis or in portal hypertension in the context of liver cirrhosis. An accessory spleen is also often found. Sometimes after splenectomy, small remains are found which may later hypertrophy. Where focal changes are concerned, we differentiate between various forms of spleen cysts, lymphomas, metastases, and benign tumors.

A pH-sensitive eosin-block copolymer delivers proteins intracellularly.

There is great interest in developing strategies to deliver proteins into the cytoplasm of cells. We report here a PEG-poly-eosin block copolymer (PEG-pEosin) that can encapsulate proteins and release them in active form under mildly acidic conditions. A PEG-pEosin formulation composed of Cre and the endosomolytic protein LLO efficiently performed gene editing in cells and in the brains of mice after an intracranial injection.

[CME Sonography 94/Answers: Adrenal Glands]. / CME-Sonografie 94/Antworten: Nebennieren.

CME Sonography 94/Answers: Adrenal Glands Abstract. Adrenal glands are often not examined in abdominal ultrasound because it is thought that the adrenal glands cannot be visualized. The normal right adrenal gland can be represented in most cases and the left one very often. Sonography makes an important contribution to the diagnosis of adrenal diseases. In addition to bleeding and tuberculosis, there are benign tumors (adrenal adenomas), pheochromocytomas, and adrenal carcinomas. There are also adrenal metastases and lymphomas.

CME-Sonografie 94: Nebennieren.

Adrenal glands are often not examined in abdominal ultrasound because it is thought that the adrenal glands cannot be visualized. The normal right adrenal gland can be represented in most cases and the left one very often. Sonography makes an important contribution to the diagnosis of adrenal diseases. In addition to bleeding and tuberculosis, there are benign tumors (adrenal adenomas), pheochromocytomas, and adrenal carcinomas. There are also adrenal metastases and lymphomas.