Somatic Mutations During an Immune Response in Xenopus Tadpoles

The tadpole B-cell repertoire is less diverse than that of the adult frog; their antibodies are of lower affinity and are less heterogenous. In order to determine whether this difference is due to a lack of or a reduced rate of somatic hypermutation, we analyzed and compared cDNA sequences utilizing VH1 elements with germline counterparts in isogenic LG7 tadpoles during an immune response. Indeed, tadpole VH1 sequences contained somatic mutations. There were zeo to 5 mutations per sequence, all single base-point mutations, with the high ratio of GC to AT base-pair alterations similar to that observed in adult frogs

Sequences of Cμ and the VH1 Family in LG7, a Clonable Strain of Xenopus, Homozygous for the Immunoglobulin Loci

Twenty-eight heavy-chain variable (VH1) region genes and the immunoglobulin (IgM) heavy-chain constant region of an isogenic Xenopus hybrid, X. laevis/X, gilli, LG7, have been sequenced. The LG7 clone represents the first Xenopus hybrid that is homozygous for the IgH locus. The VH1 family was specifically investigated because VH1 genes are used by the antibodies produced during the Xenopus antidinitrophenol (DNP) response, These VH1 germ-line sequences establish a so-called ”dictionary— that is available for studying somatic hypermutational mechanisms in immunized frogs

Spinal manipulation frequency and dosage effects on clinical and physiological outcomes: a scoping review

Introduction: The burden of musculoskeletal disorders increases every year, with low back and neck pain being the most frequently reported conditions for seeking manual therapy treatment. In recent years, manual therapy research has begun exploring the dose-response relationship between spinal manipulation treatment characteristics and both clinical and physiological response to treatment. Objective: The purpose of this scoping review was to identify and appraise the current state of scientific knowledge regarding the effects of spinal manipulation frequency and dosage on both clinical and physiological responses. Methods: A scoping review was conducted to identify all available studies pertaining to our research question. Retrieved papers were screened using a 2-phase method, a selective sorting with titles and abstracts. Potentially relevant studies were read, and data was extracted for all included studies. Randomized control trials were assessed using the Cochrane Risk of Bias Tool for quality assessment. Results: The search yielded 4854 publications from which 32 were included for analysis. Results were sorted by dosage or frequency outcomes, and divided into human or animal studies. Animal studies mainly focused on dosage and evaluated physiological outcomes only. Studies investigating spinal manipulation dosage effects involved both human and animal research, and showed that varying thrust forces, or thrust durations can impact vertebral displacement, muscular response amplitude or muscle spindle activity. Risk of bias analysis indicated only two clinical trials assessing frequency effects presented a low risk of bias. Although trends in improvement were observed and indicated that increasing the number of SM visits in a short period of time (few weeks) decreased pain and improve disability, the differences between the studied treatment frequencies, were often not statistically significant and therefore not clinically meaningful. Conclusion: The results of this study showed that SM dosage and frequency effects have been mostly studied over the past two decades. Definitions for these two concepts however differ across studies. Overall, the results showed that treatment frequency does not significantly affect clinical outcomes during and following a SM treatment period. Dosage effects clearly influence short-term physiological responses to SM treatment, but relationships between these responses and clinical outcomes remains to be investigated. © 2019 The Author(s)

SARS-CoV-2 infects epithelial cells of the blood-cerebrospinal fluid barrier rather than endothelial cells or pericytes of the blood-brain barrier.

BACKGROUND As a consequence of SARS-CoV-2 infection various neurocognitive and neuropsychiatric symptoms can appear, which may persist for several months post infection. However, cell type-specific routes of brain infection and underlying mechanisms resulting in neuroglial dysfunction are not well understood. METHODS Here, we investigated the susceptibility of cells constituting the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB) of the choroid plexus (ChP) to SARS-CoV-2 infection using human induced pluripotent stem cell (hiPSC)-derived cellular models and a ChP papilloma-derived epithelial cell line as well as ChP tissue from COVID-19 patients, respectively. RESULTS We noted a differential infectibility of hiPSC-derived brain microvascular endothelial cells (BMECs) depending on the differentiation method. Extended endothelial culture method (EECM)-BMECs characterized by a complete set of endothelial markers, good barrier properties and a mature immune phenotype were refractory to SARS-CoV-2 infection and did not exhibit an activated phenotype after prolonged SARS-CoV-2 inoculation. In contrast, defined medium method (DMM)-BMECs, characterized by a mixed endothelial and epithelial phenotype and excellent barrier properties were productively infected by SARS-CoV-2 in an ACE2-dependent manner. hiPSC-derived brain pericyte-like cells (BPLCs) lacking ACE2 expression were not susceptible to SARS-CoV-2 infection. Furthermore, the human choroid plexus papilloma-derived epithelial cell line HIBCPP, modeling the BCSFB was productively infected by SARS-CoV-2 preferentially from the basolateral side, facing the blood compartment. Assessment of ChP tissue from COVID-19 patients by RNA in situ hybridization revealed SARS-CoV-2 transcripts in ChP epithelial and ChP stromal cells. CONCLUSIONS Our study shows that the BCSFB of the ChP rather than the BBB is susceptible to direct SARS-CoV-2 infection. Thus, neuropsychiatric symptoms because of COVID-19 may rather be associated with dysfunction of the BCSFB than the BBB. Future studies should consider a role of the ChP in underlying neuropsychiatric symptoms following SARS-CoV-2 infection