An essential role for the Zn2+ transporter ZIP7 in B cell development.

Despite the known importance of zinc for human immunity, molecular insights into its roles have remained limited. Here we report a novel autosomal recessive disease characterized by absent B cells, agammaglobulinemia and early onset infections in five unrelated families. The immunodeficiency results from hypomorphic mutations of SLC39A7, which encodes the endoplasmic reticulum-to-cytoplasm zinc transporter ZIP7. Using CRISPR-Cas9 mutagenesis we have precisely modeled ZIP7 deficiency in mice. Homozygosity for a null allele caused embryonic death, but hypomorphic alleles reproduced the block in B cell development seen in patients. B cells from mutant mice exhibited a diminished concentration of cytoplasmic free zinc, increased phosphatase activity and decreased phosphorylation of signaling molecules downstream of the pre-B cell and B cell receptors. Our findings highlight a specific role for cytosolic Zn2+ in modulating B cell receptor signal strength and positive selection.

Natural antisense transcripts as versatile regulators of gene expression.

Long non-coding RNAs (lncRNAs) are emerging as a major class of gene products that have central roles in cell and developmental biology. Natural antisense transcripts (NATs) are an important subset of lncRNAs that are expressed from the opposite strand of protein-coding and non-coding genes and are a genome-wide phenomenon in both eukaryotes and prokaryotes. In eukaryotes, a myriad of NATs participate in regulatory pathways that affect expression of their cognate sense genes. Recent developments in the study of NATs and lncRNAs and large-scale sequencing and bioinformatics projects suggest that whether NATs regulate expression, splicing, stability or translation of the sense transcript is influenced by the pattern and degrees of overlap between the sense-antisense pair. Moreover, epigenetic gene regulatory mechanisms prevail in somatic cells whereas mechanisms dependent on the formation of double-stranded RNA intermediates are prevalent in germ cells. The modulating effects of NATs on sense transcript expression make NATs rational targets for therapeutic interventions.

Age-Specific Resurgence in Invasive Pneumococcal Disease Incidence in the COVID-19 Pandemic Era and Its Association With Respiratory Virus and Pneumococcal Carriage Dynamics: A Time-Series Analysis.

Using multiple national surveillance systems, we found an increase in the incidence of invasive pneumococcal disease during after the relaxation of non-pharmaceutical interventions against COVID-19, which strongly varied by age. Age groups with higher incidence of respiratory syncytial virus and influenza also experienced higher increase in invasive pneumococcal disease incidence, with no change in pneumococcal carriage.

XPR1: a regulator of cellular phosphate homeostasis rather than a Pi exporter.

Phosphate (Pi) is an essential nutrient, and its plasma levels are under tight hormonal control. Uphill transport of Pi into cells is mediated by the two Na-dependent Pi transporter families SLC34 and SLC20. The molecular identity of a potential Pi export pathway is controversial, though XPR1 has recently been suggested by Giovannini and coworkers to mediate Pi export. We expressed XPR1 in Xenopus oocytes to determine its functional characteristics. Xenopus isoforms of proteins were used to avoid species incompatibility. Protein tagging confirmed the localization of XPR1 at the plasma membrane. Efflux experiments, however, failed to detect translocation of Pi attributable to XPR1. We tested various counter ions and export medium compositions (pH, plasma) as well as potential protein co-factors that could stimulate the activity of XPR1, though without success. Expression of truncated XPR1 constructs and individual domains of XPR1 (SPX, transmembrane core, C-terminus) demonstrated downregulation of the uptake of Pi mediated by the C-terminal domain of XPR1. Tethering the C-terminus to the transmembrane core changed the kinetics of the inhibition and the presence of the SPX domain blunted the inhibitory effect. Our observations suggest a regulatory role of XPR1 in cellular Pi handling rather than a function as Pi exporter. Accordingly, XPR1 senses intracellular Pi levels via its SPX domain and downregulates cellular Pi uptake via the C-terminal domain. The molecular identity of a potential Pi export protein remains therefore elusive.

Widespread formation of double-stranded RNAs in testis.

The testis transcriptome is highly complex and includes RNAs that potentially hybridize to form double-stranded RNA (dsRNA). We isolated dsRNA using the monoclonal J2 antibody and deep-sequenced the enriched samples from testes of juvenile Dicer1 knockout mice, age-matched controls, and adult animals. Comparison of our data set with recently published data from mouse liver revealed that the dsRNA transcriptome in testis is markedly different from liver: In testis, dsRNA-forming transcripts derive from mRNAs including promoters and immediate downstream regions, whereas in somatic cells they originate more often from introns and intergenic transcription. The genes that generate dsRNA are significantly expressed in isolated male germ cells with particular enrichment in pachytene spermatocytes. dsRNA formation is lower on the sex (X and Y) chromosomes. The dsRNA transcriptome is significantly less complex in juvenile mice as compared to adult controls and, possibly as a consequence, the knockout of Dicer1 has only a minor effect on the total number of transcript peaks associated with dsRNA. The comparison between dsRNA-associated genes in testis and liver with a reported set of genes that produce endogenous siRNAs reveals a significant overlap in testis but not in liver. Testis dsRNAs also significantly associate with natural antisense genes-again, this feature is not observed in liver. These findings point to a testis-specific mechanism involving natural antisense transcripts and the formation of dsRNAs that feed into the RNA interference pathway, possibly to mitigate the mutagenic impacts of recombination and transposon mobilization.

New French height velocity growth charts: An innovative big-data approach based on routine measurements.

AIM: Height velocity is considered a key auxological tool to monitor growth, but updated height velocity growth charts are lacking. We aimed to derive new French height velocity growth charts by using a big-data approach based on routine measurements. METHODS: We extracted all growth data of children aged 1 month-18 years from the electronic medical records of 42 primary care physicians, between 1 January 1990 and 8 February 2018, throughout the French metropolitan territory. We derived annual and biannual height velocity growth charts until age 15 years by using the Lambda-Mu-Sigma method. These new growth charts were compared to the 1979 French and 2009 World Health Organisation (WHO) ones. RESULTS: New height velocity growth charts were generated with 193 124 and 209 221 annual and biannual values from 80 204 and 87 260 children, respectively, and showed good internal fit. Median curves were close to the 1979 French or 2009 WHO ones, but SD curves displayed important differences. Similar results were found with the biannual height velocity growth charts. CONCLUSION: We produced new height velocity growth charts until age 15 years by using a big-data approach applied to measurements routinely collected in clinical practice. These updated growth charts could help optimise growth-monitoring performance.

Comparative study on tertiary contacts and folding of RNase P RNAs from a psychrophilic, a mesophilic/radiation-resistant, and a thermophilic bacterium.

In most bacterial type A RNase P RNAs (P RNAs), two major loop-helix tertiary contacts (L8-P4 and L18-P8) help to orient the two independently folding S- and C-domains for concerted recognition of precursor tRNA substrates. Here, we analyze the effects of mutations in these tertiary contacts in P RNAs from three different species: (i) the psychrophilic bacterium Pseudoalteromonas translucida (Ptr), (ii) the mesophilic radiation-resistant bacterium Deinococcus radiodurans (Dra), and (iii) the thermophilic bacterium Thermus thermophilus (Tth). We show by UV melting experiments that simultaneous disruption of these two interdomain contacts has a stabilizing effect on all three P RNAs. This can be inferred from reduced RNA unfolding at lower temperatures and a more concerted unfolding at higher temperatures. Thus, when the two domains tightly interact via the tertiary contacts, one domain facilitates structural transitions in the other. P RNA mutants with disrupted interdomain contacts showed severe kinetic defects that were most pronounced upon simultaneous disruption of the L8-P4 and L18-P8 contacts. At 37°C, the mildest effects were observed for the thermostable Tth RNA. A third interdomain contact, L9-P1, makes only a minor contribution to P RNA tertiary folding. Furthermore, D. radiodurans RNase P RNA forms an additional pseudoknot structure between the P9 and P12 of its S-domain. This interaction was found to be particularly crucial for RNase P holoenzyme activity at near-physiological Mg2+ concentrations (2 mM). We further analyzed an exceptionally stable folding trap of the G,C-rich Tth P RNA.

Midterm clinical outcome of uncemented short-stem reversed shoulder arthroplasty.

INTRODUCTION: While the incidence of reverse total shoulder arthroplasty (rTSA) is increasing constantly, newer implants with designs other than the classic Grammont geometry are gaining importance. More anatomic inclination angles and lateralization are supposed to have a positive impact on clinical results and complication rates. Presentation of midterm results therefore is important to support these assumptions. The aim of this study was to report the midterm clinical outcome of primary rTSA with an uncemented humeral short-stem prosthesis (USSP) with a humeral inclination angle of 145° and the analysis of different variables on the outcome. METHODS: This is a retrospective study of all patients with primary rTSA using an USSP and a combined humeral inclination angle of 145° (Ascend™ flex, Stryker) with a minimum clinical follow-up of 2 years. The implant combines a 132.5° inclination for the humeral stem with an additional 12.5° for the polyethylene inlay. Primary outcomes were patient-reported outcome measures: ASES score, simple shoulder test (SST) and subjective shoulder value (SSV). Secondary outcomes were complication and revision rates. We analyzed different variables: preoperatively gender, age, indication for surgery and status of rotator cuff. Also, the glenoid morphology was classified according to Walch and a proximal humerus cortical bone thickness measurement (CBT avg) of 6 mm was used as a threshold for osteoporosis. Postoperatively, we analyzed different radiologic parameters: filling ratio, distalization and lateralization angles according to Boutsiadis. RESULTS: A total of 84 out of 99 (85%) patients with a mean FU of 46.7 months (range 24-80 months) could be included: 62 women and 22 men with a mean age of 74.7 years. Mean ASES score significantly increased from 47 preoperatively to 85.8 at the last follow-up (p = 0.001). The postoperative SST reached an average of 65.3 and the mean SSV was 83%. None of the variable parameters analyzed could be identified as a risk factor for a lower outcome defined as a SSV < 70. Three patients (3.6%) had a complication: one incomplete lower plexus lesion, one dislocation and one major hematoma. Surgical revision was needed in two cases (2.4%). CONCLUSION: The midterm clinical outcome of primary reverse total shoulder arthroplasty (rTSA) with an uncemented humeral short stem and a humeral inclination angle of 145° showed good-to-excellent results with a low complication and revision rate independent from a wide range of pre- and postoperative variables. PROMs are comparable to those reported for anatomic TSA with a low complication rate, different to historical studies especially with the Grammont design. LEVEL OF EVIDENCE: Treatment study, Level IV.

Cardiovascular Regulation During Acute Gravitational Changes with Exhaling on Exertion.

During gravitational changes or changes in the direction of action in relation to the body, fluid displacements can be observed. In special cases different breathing maneuvers (e. g., exhaling on exertion; Ex-Ex) are used to counteract acute fluid shifts. Both factors have a significant impact on cardiovascular regulation. Eight healthy male subjects were tested on a tilt seat, long arm human centrifuge, and parabolic flight. The work aims to investigate the effect of exhaling on exertion on the cardiovascular regulation during acute gravitational changes compared to normal breathing. Possible interactions and differences between conditions (Ex-Ex, normal breathing) for the parameters V'O 2 , V' E , HR, and SV were analysed over a 40 s period by a three-way ANOVA. Significant (p≤0.05) effects for all main factors and interactions between condition and time as well as maneuver and time were found for all variables. The exhaling on exertion maneuver had a significant influence on the cardiovascular response during acute gravitational and positional changes. For example, the significant increase of V'O2 at the end of the exhalation on exertion maneuver indicates an increased lung circulation as a result of the maneuver.