Calmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii

Toxoplasma gondii contains an expanded number of calmodulin (CaM)-like proteins whose functions are poorly understood. Using a combination of CRISPR/Cas9-mediated gene editing and a plant-like auxin-induced degron (AID) system, we examined the roles of three apically localized CaMs. CaM1 and CaM2 were individually dispensable, but loss of both resulted in a synthetic lethal phenotype. CaM3 was refractory to deletion, suggesting it is essential. Consistent with this prediction auxin-induced degradation of CaM3 blocked growth. Phenotypic analysis revealed that all three CaMs contribute to parasite motility, invasion, and egress from host cells, and that they act downstream of microneme and rhoptry secretion. Super-resolution microscopy localized all three CaMs to the conoid where they overlap with myosin H (MyoH), a motor protein that is required for invasion. Biotinylation using BirA fusions with the CaMs labeled a number of apical proteins including MyoH and its light chain MLC7, suggesting they may interact. Consistent with this hypothesis, disruption of MyoH led to degradation of CaM3, or redistribution of CaM1 and CaM2. Collectively, our findings suggest these CaMs may interact with MyoH to control motility and cell invasion

Osteo-Chondroprogenitor–Specific Deletion of the Selenocysteine tRNA Gene, Trsp, Leads to Chondronecrosis and Abnormal Skeletal Development: A Putative Model for Kashin-Beck Disease

Kashin-Beck disease, a syndrome characterized by short stature, skeletal deformities, and arthropathy of multiple joints, is highly prevalent in specific regions of Asia. The disease has been postulated to result from a combination of different environmental factors, including contamination of barley by mold mycotoxins, iodine deficiency, presence of humic substances in drinking water, and, importantly, deficiency of selenium. This multifunctional trace element, in the form of selenocysteine, is essential for normal selenoprotein function, including attenuation of excessive oxidative stress, and for the control of redox-sensitive molecules involved in cell growth and differentiation. To investigate the effects of skeletal selenoprotein deficiency, a Cre recombinase transgenic mouse line was used to trigger Trsp gene deletions in osteo-chondroprogenitors. Trsp encodes selenocysteine tRNA[Ser]Sec, required for the incorporation of selenocysteine residues into selenoproteins. The mutant mice exhibited growth retardation, epiphyseal growth plate abnormalities, and delayed skeletal ossification, as well as marked chondronecrosis of articular, auricular, and tracheal cartilages. Phenotypically, the mice thus replicated a number of the pathological features of Kashin-Beck disease, supporting the notion that selenium deficiency is important to the development of this syndrome

Transplante lobar experimental em suínos: enxerto proporcional na disparidade entre receptor e doador Experimental lobar transplantation in swine: proportional graft in the discrepancy between donor and recipient

Objetivo - Perante a crítica falta de orgãos disponíveis para transplante, a doação intervivos de lobos ou segmentos pulmonares é possível e necessária no transplante pulmonar pediátrico. Na grande disparidade de tamanho do doador adulto e a criança pequena receptora, o lobo médio ou segmento pulmonar representa um enxerto de restrito leito vascular. Este estudo experimental testa a hipótese de que esse tipo de enxerto pode desenvolver hipertensão pulmonar no animal receptor, ao final do período de crescimento. Métodos - Foi utilizado um modelo de sobrevivência de transplante pulmonar à esquerda em leitões. Constituíram-se três grupos de comparação: I (n = 4) - transplante de lobo superior de doador adulto, enxerto proporcional ao receptor, mas irrigado por apenas dois ramos arteriais; II (n = 5) - transplante de lobo inferior de doador adulto, enxerto desproporcional ao receptor e com amplo leito vascular; III (n = 6) - transplante de pulmão imaturo de leitão doador proporcionado. Os animais transplantados tiveram a função do enxerto pulmonar estudada ao final de 3 meses, quando completaram o período de crescimento. Resultados - A pressão da artéria pulmonar do enxerto do grupo I (51,8 ± 2,1mmHg) foi mais elevada do que no grupo II (40,4 ± 2,5mmHg) e do que no grupo III (34,8 ± 1,5mmHg), atingindo significância estatística (p = 0,0003). Conclusões - O enxerto lobar proporcional ao receptor, mas de leito vascular restrito, teve desempenho hemodinâmico comprometido no animal em crescimento. Esses dados sugerem que a proporcionalidade do enxerto não deve ser prioritária e, antes, ser secundária a um adequado leito vascular do enxerto.<br>Background - The critical donor shortage in pediatric pulmonary transplantation has prompted lobar transplantation from living-related. However, in the case of great size discrepancy between the adult donor and the small child recipient, a pulmonary segment or medium lobe represents grafts with restricted vascular bed. The authors hypothesized that this type of graft may develop pulmonary hypertension in the recipient by the end of the growth period. Methods - This hypothesis was investigated in a porcine survival model of lung transplantation in piglets. There were three groups for comparison purposes: I (n = 4) - transplantation of the upper lobe from an adult donor, graft being proportional to the recipient but irrigated by two arterial rami only; II (n = 5) - transplantation of the lower lobe from an adult donor, graft being oversized to the recipient and having adequate vascular bed; III (n = 6) - transplantation of immature lung, from matched-sized donor. Graft function was studied three months after the transplantation, when the growth period was completed. Results - The pulmonary artery pressure of grafts in group I (51.8 ± 2.1 mmHg) was increased compared to that of group II (40.4 ± 2.5 mmHg) and of group III (34.8 ± 1.5 mmHg), reaching statistical significance (p = 0.0003). Conclusions - The lobar graft proportional to the recipient, with restricted vascular bed, had hampered hemodynamic performance in the growing animal. These results suggest that graft proportionality should be secondary to an adequate vascular bed