Genome editing in plants using CRISPR type I-D nuclease

Genome editing in plants has advanced greatly by applying the clustered regularly interspaced short palindromic repeats (CRISPRs)-Cas system, especially CRISPR-Cas9. However, CRISPR type I—the most abundant CRISPR system in bacteria—has not been exploited for plant genome modification. In type I CRISPR-Cas systems, e.g., type I-E, Cas3 nucleases degrade the target DNA in mammals. Here, we present a type I-D (TiD) CRISPR-Cas genome editing system in plants. TiD lacks the Cas3 nuclease domain; instead, Cas10d is the functional nuclease in vivo. TiD was active in targeted mutagenesis of tomato genomic DNA. The mutations generated by TiD differed from those of CRISPR/Cas9; both bi-directional long-range deletions and short indels mutations were detected in tomato cells. Furthermore, TiD can be used to efficiently generate bi-allelic mutant plants in the first generation. These findings indicate that TiD is a unique CRISPR system that can be used for genome engineering in plants

Recurrence of Proliferative Glomerulonephritis with Monoclonal Immunoglobulin G Deposits with a Striated Ultrastructure

This is the peer-reviewed but unedited manuscript version of the following article: Nephron 2020;144(suppl 1):43–48 (DOI: 10.1159/000512330)]. The final, published version is available at http://www.karger.com/?doi=10.1159/000512330

Adverse events following immunization of elderly with COVID-19 inactivated virus vaccine (CoronaVac) in Southeastern Brazil: an active surveillance study

Healthcare workers, the elderly and other vulnerable populations were the first to receive COVID-19 vaccines in public health programs. There were few vaccine safety data available on the elderly. This observational study aimed to evaluate the inactivated vaccine (CoronaVac) safety in the elderly, at the beginning of the vaccination program, in Sao Paulo city, Brazil. The elderly people that received CoronaVac at the Reference Center for Special Immunobiologicals (CRIE) or at home, administered by the Interdisciplinary Home Care Team (NADI) of the Hospital das Clinicas were invited to participate in this phase 4 observational study. The vaccination schedule included two CoronaVac doses 28 days apart. The information on solicited and unsolicited adverse events following immunization were collected by phone calls on days 4 and 8 after each vaccine dose. We enrolled 158 adults aged 65 to 101 years (mean of 84.1 years); 63.9% were females and 95.6% had chronic conditions, 21.5% had moderate or severe impairment in daily living activities; 34.2% were pre-frail and 19.6% were frail. We were able to contact 95.6% and 91.6% of the vaccinated people, after the first and second doses, respectively; 31.8% and 23.4% of the contacted participants reported some adverse events (AE) following the first and second doses, respectively. Pain at the injection site, fatigue, myalgia and headaches were the most frequent solicited AE. Most AE were mild to moderate. There were eight severe adverse events, but none of them were considered related to the vaccine. The CoronaVac was safe and well tolerated by these adults of advanced age with frailty and comorbidities

Humoral immune response in experimental ddY mice Paracoccidioidomycosis

A paracoccidioidomicose (PCM) é uma micose sistêmica, restrita à América Latina, com maior incidência no Brasil. O camundongo ddY tem sido empregado como modelo murino de PCM e, no entanto, não há informações a respeito da resposta imune desse animal frente à infecção. O presente estudo tem como objetivo avaliar a resposta imune humoral específica para o principal antígeno, gp43, do fungo Paracoccidioides brasiliensis, em camundongos ddY infectados com a cepa virulenta Pb 18. Foram realizadas análises da antigenemia e histopatológico em vários órgãos e em diferentes tempos pós-infecção. Os resultados obtidos demonstraram aumento nos níveis de IgG anti-gp43 nos dias 14, 17, 21, 24, 28 e 56 pós-infecção e aumento no nível de gp43 solúvel aos 28 dias pós-infecção. As células fúngicas foram detectadas em todos os órgãos analisados (cérebro, coração, pulmão, fígado, baço e rim) e em todos os períodos. As lesões granulomatosas tornaramse predominantes 14 dias pós-infecção. Os resultados evidenciaram que o camundongo ddY produz resposta imune humoral frente ao principal antígeno de P. brasiliensis, apresentando-se elevado até 56 dias pósinfecção. A redução do nível de gp43 solúvel na fase crônica, supostamente devido ao início do controle da infecção, requer estudos complementares adicionais.Paracoccidioidomycosis (PCM) is a systemic mycosis, restrict to Latin America, with higher incidence in Brazil. ddY mice have been used as experimental PCM model, although there is no data regarding immune response. The aim of the present study was evaluated specific humoral response against the main specific antigen of the fungal Paracoccidioides brasiliensis, the gp43, in ddY mice infected with virulent Pb 18. Antigenemia analysis and histophatological exam in several organs were performed in different time postinfection The results showed increased levels of anti-gp43 IgG on days 14, 17, 21, 24, 28 and 56 post-infection and increased levels of soluble gp-43 on day 28 post-infection. The fungal cells were detected in all organs analyzed (brain, heart, lung, liver, spleen and kidney) in all investigated periods. The granulomatous lesions became predominant 14 days after infection. The results evidence that ddY mice produce humoral immune response to main P. brasiliensis antigen, with high levels until 56 days after infection. Further studies are needed to show that reduction of soluble gp43 in chronic phase correlates with infection control

Bioactive micropatterning of apatite immobilizing cell adhesion protein by laser-induced forward transfer with a shock absorber

The additive patterning of apatite with good biocompatibility and osteoconductivity is a useful technique for the production and surface functionalization of biomaterials. We developed this technique through our laser-induced forward transfer (LIFT) process using a laser-absorbing sacrificial layer in combination with a shock-absorbing polydimethylsiloxane (PDMS) receiver. With the PDMS shock-absorbing function, even the brittle apatite and that immobilizing the cell adhesion protein fibronectin (Fn-apatite) were successfully transferred and micropatterned while maintaining their dense, filmy state. The laser pulse energy effect was investigated, leading to the optimum energy range just above the transfer threshold. The apatite and Fn-apatite micropatterns exhibited superior cytocompatibility compared to the PDMS surface, and could potentially be used for cellular micromanipulation. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreemen