Counteracting the Common Shwachman–Diamond Syndrome-Causing SBDS c.258+2T>C Mutation by RNA Therapeutics and Base/Prime Editing

Shwachman-Diamond syndrome (SDS) represents one of the most common inherited bone marrow failure syndromes and is mainly caused by SBDS gene mutations. Only supportive treatments are available, with hematopoietic cell transplantation required when marrow failure occurs. Among all causative mutations, the SBDS c.258+2T>C variant at the 5 ' splice site (ss) of exon 2 is one of the most frequent. Here, we investigated the molecular mechanisms underlying aberrant SBDS splicing and showed that SBDS exon 2 is dense in splicing regulatory elements and cryptic splice sites, complicating proper 5 ' ss selection. Studies ex vivo and in vitro demonstrated that the mutation alters splicing, but it is also compatible with tiny amounts of correct transcripts, which would explain the survival of SDS patients. Moreover, for the first time for SDS, we explored a panel of correction approaches at the RNA and DNA levels and provided experimental evidence that the mutation effect can be partially counteracted by engineered U1snRNA, trans-splicing, and base/prime editors, ultimately leading to correctly spliced transcripts (from barely detectable to 2.5-5.5%). Among them, we propose DNA editors that, by stably reverting the mutation and potentially conferring positive selection to bone-marrow cells, could lead to the development of an innovative SDS therapy

A Feasibility Study on Indoor Localization and Multi-person Tracking Using Sparsely Distributed Camera Network with Edge Computing

Camera-based activity monitoring systems are becoming an attractive solution for smart building applications with the advances in computer vision and edge computing technologies. In this paper, we present a feasibility study and systematic analysis of a camera-based indoor localization and multi-person tracking system implemented on edge computing devices within a large indoor space. To this end, we deployed an end-to-end edge computing pipeline that utilizes multiple cameras to achieve localization, body orientation estimation and tracking of multiple individuals within a large therapeutic space spanning $1700m^2$, all while maintaining a strong focus on preserving privacy. Our pipeline consists of 39 edge computing camera systems equipped with Tensor Processing Units (TPUs) placed in the indoor space's ceiling. To ensure the privacy of individuals, a real-time multi-person pose estimation algorithm runs on the TPU of the computing camera system. This algorithm extracts poses and bounding boxes, which are utilized for indoor localization, body orientation estimation, and multi-person tracking. Our pipeline demonstrated an average localization error of 1.41 meters, a multiple-object tracking accuracy score of 88.6\%, and a mean absolute body orientation error of 29\degree. These results shows that localization and tracking of individuals in a large indoor space is feasible even with the privacy constrains

Extracellular vesicles in human preterm colostrum inhibit infection by human cytomegalovirus in vitro

Breast milk is a complex biofluid that nourishes infants, supports their growth and protects them from diseases. However, at the same time, breastfeeding is a transmission route for human cytomegalovirus (HCMV), with preterm infants being at a great risk of congenital disease. The discrepancy between high HCMV transmission rates and the few reported cases of infants with severe clinical illness is likely due to the protective effect of breast milk. The aim of this study was to investigate the anti-HCMV activity of human preterm colostrum and clarify the role of colostrum-derived extracellular vesicles (EVs). Preterm colostrum samples were collected and the EVs were purified and characterized. The in vitro anti-HCMV activity of both colostrum and EVs was tested against HCMV, and the viral replication step inhibited by colostrum-purified EVs was examined. We investigated the putative role EV surface proteins play in impairing HCMV infection using shaving experiments and proteomic analysis. The obtained results confirmed the antiviral action of colostrum against HCMV and demonstrated a remarkable antiviral activity of colostrum-derived EVs. Furthermore, we demonstrated that EVs impair the attachment of HCMV to cells, with EV surface proteins playing a role in mediating this action. These findings contribute to clarifying the mechanisms that underlie the protective role of human colostrum against HCMV infection

Više-fragmentna tvorba u reakcijama 32S+58,64Ni NA 11 A MeV

The characteristic features of the 32S+58,64Ni reaction at 11 A MeV have been investigated to evidence the possible rise of multi-fragmentation processes at low excitation energies. The importance of such phenomena consists in the fact that they could represent the signature of a nuclear phase transition from a liquid to a gas region. Evidence of multi-fragment production is displayed by the present data; even if the yield of such events is compatible with the predictions of statistical models, the partition of the mass of the decaying system cannot be easily reproduced. Some preliminary indications of isospin effects are suggested by looking at the differences between the two reacting systems.Istraživali smo značajke reakcije 32S+ 58,64Ni na 11 A MeV radi pronalaženja mogućeg porasta više-fragmentnih procesa na nižim uzbudnim energijama. Važnost se tih pojava sastoji u tome što bi one mogle predstavljati znakove nuklearnog faznog prijelaza od tekućeg u plinsko stanje. Znakovi tvorbe više fragmenata vide se u dobivenim podacima; no čak ako je i prinos tih događaja u skladu s predviđanjima statističkog modela, razdioba mase sustava koji se raspada ne uspijeva se objasniti. Usporedba dviju reakcija ukazuje na moguće izospinske efekte

Temperature measurement of fragment emitting systems in Au+Au 35 MeV/nucleon collisions

We report on the results of experiments performed to investigate the Au1Au 35 MeV/nucleon reaction. The reaction products generated in the disassembly of the unique source formed in central collisions and those coming from the decay of the quasiprojectile in peripheral and midperipheral ones ~five different impact parameters! were identified through a careful data selection based on the study of energy and angular distributions. The excitation energies of the fragment sources have been extracted through a calorimetric method and by means of a comparison with model calculations. The nuclear temperatures of these decaying systems have been measured from the relative isotopic abundances and, also for central collisions, from the relative populations of excited states. The temperatures of the quasiprojectile disassembling systems are slowly increasing going towards smaller impact parameter. The relationship between temperature and excitation energy seems to be almost independent of the characteristics of the emitting source. The extracted caloric curve shows a slow monotonic increase with increasing excitation energy. A comparison with data derived from Au fragmentation at much higher incident energies is discussed. @S0556-2813~98!04408-2

CLUSTER ANALYSIS IN SEASONAL DECIDUOUS FOREST REMNANTS

Floresta Estacional Decidual, caracterizado como um enclave c\uf4ncavo, em terreno com declividade acentuada e solo arenoso, entre campos caracter\uedsticos da regi\ue3o. As esp\ue9cies arb\uf3reas e arbustivas com CAP (circunfer\ueancia a altura de 1,3 m do solo) 65 15 cm foram amostradas em duas classes de tamanho. Na amostragem dos indiv\uedduos da Classe I (CAP 65 30 cm) foram utilizadas 14 parcelas de 20 x 100 m, divididas em subparcelas (10 x 10 m). Os indiv\uedduos da Classe II (15 64 CAP < 30 cm) foram observados em 70 subparcelas. Para an\ue1lise de agrupamentos foi utilizado o programa TWINSPAN, que indicou dois grupos (G1 e G2) bem definidos na Classe I e um grupo relativamente homog\ueaneo na Classe II (sub-bosque). No grupo G1 ocorreram as esp\ue9cies indicadoras Trichilia claussenii , Cupania vernalis e Crysophyllum marginatum ; e no grupo G2 Luehea divaricata e Sebastiania commersoniana . Em rela\ue7\ue3o \ue0 estrutura horizontal, essas esp\ue9cies estiveram entre as tr\ueas mais importantes nos respectivos grupos. No sub-bosque, as esp\ue9cies com maior valor de import\ue2ncia foram Actinostemon concolor , Trichilia claussenii, Trichilia elegans , Eugenia rostrifolia e Sorocea bonplandii . Essas esp\ue9cies, por predominarem na vegeta\ue7\ue3o arb\uf3rea, apresentam papel fundamental na estabiliza\ue7\ue3o de encostas, considerando que est\ue3o adaptadas \ue0s \ue1reas com declividade acentuada e solos rasos, destacando-se Cupania vernalis e Trichilia claussenii em ambientes menos alterados e Luehea divaricata e Sebastiania commersoniana em ambientes mais suscept\uedveis \ue0s interfer\ueancias.This study intended to analyze the presence of clusters in the vegetation of Seasonal Deciduous Forest remnants, characterized as a concave enclave, on steep sandy soil, among the region\u2019s characteristic fields. Shrub and tree specimens with CBH (circumference at breast height) 65 15 cm were sampled in two size classes. In class I (CBH 65 30 cm) specimen sampling, we used fourteen 20 x 100 m plots, divided into sub-plots (10 x 10 m). Class II (15 64 CBH < 30 cm) specimens were observed in 70 sub-portions. We used the TWINSPAN program to analyze the clusters, which indicated two well-defined clusters in class I (G1 and G2) and one relatively homogenous cluster in class II (understorey). There were Trichilia claussenii , Cupania vernalis and Crysophyllum marginatum indicator species in the G1 cluster; and Luehea divaricata and Sebastiania commersoniana in the G2 cluster. Regarding to horizontal structure, these species were among the three most important ones in both clusters. In the understorey, the most important species were Actinostemon concolor , Trichilia claussenii, Trichilia elegans , Eugenia rostrifolia and Sorocea bonplandii . Since these species prevail in the tree vegetation, they are fundamental to stabilize hillsides, considering that they are adapted to steep and shallow soil areas, pointing out the Cupania vernalis and Trichilia claussenii in less modified environments and the Luehea divaricata and Sebastiania commersoniana in environments more susceptible to interference