Current insights and advances into plant male sterility: new precision breeding technology based on genome editing applications

Plant male sterility (MS) represents the inability of the plant to generate functional anthers, pollen, or male gametes. Developing MS lines represents one of the most important challenges in plant breeding programs, since the establishment of MS lines is a major goal in F1 hybrid production. For these reasons, MS lines have been developed in several species of economic interest, particularly in horticultural crops and ornamental plants. Over the years, MS has been accomplished through many different techniques ranging from approaches based on cross-mediated conventional breeding methods, to advanced devices based on knowledge of genetics and genomics to the most advanced molecular technologies based on genome editing (GE). GE methods, in particular gene knockout mediated by CRISPR/Cas-related tools, have resulted in flexible and successful strategic ideas used to alter the function of key genes, regulating numerous biological processes including MS. These precision breeding technologies are less time-consuming and can accelerate the creation of new genetic variability with the accumulation of favorable alleles, able to dramatically change the biological process and resulting in a potential efficiency of cultivar development bypassing sexual crosses. The main goal of this manuscript is to provide a general overview of insights and advances into plant male sterility, focusing the attention on the recent new breeding GE-based applications capable of inducing MS by targeting specific nuclear genic loci. A summary of the mechanisms underlying the recent CRISPR technology and relative success applications are described for the main crop and ornamental species. The future challenges and new potential applications of CRISPR/Cas systems in MS mutant production and other potential opportunities will be discussed, as generating CRISPR-edited DNA-free by transient transformation system and transgenerational gene editing for introducing desirable alleles and for precision breeding strategies

Mitochondrial DNA Suggests a Western Eurasian origin for Ancient (Proto-) Bulgarians

Ancient (proto-) Bulgarians have long been thought to as a Turkic population. However, evidence found in the past three decades show that this is not the case. Until now, this evidence does not include ancient mitochondrial DNA (mtDNA) analysis. In order to fill this void, we have collected human remains from the VIII-X century AD located in three necropolises in Bulgaria: Nojarevo (Silistra region) and Monastery of Mostich (Shumen region), both in Northeast Bulgaria and Tuhovishte (Satovcha region) in Southwest Bulgaria. The phylogenetic analysis of 13 ancient DNA samples (extracted from teeth) identified 12 independent haplotypes, which we further classified into mtDNA haplogroups found in present-day European and Western Eurasian populations. Our results suggest a Western Eurasian matrilineal origin for proto-Bulgarians as well as a genetic similarity between proto- and modern Bulgarians. Our future work will provide additional data which will further clarify proto-Bulgarian origins; thereby adding new clues to current understanding of European genetic evolution

Influence of detection of pretreatment cytogenetic abnormalities on first complete remission and survival in adult acute lymphoblastic leukemia

Objective: Treatment of acute lymphoblastic leukemia (ALL) in adults focuses on the initial assessment of the prognostic relevant cytogenetic features as well as a response-guided therapy based on molecular data. We examined the importance of molecular-cytogenetic abnormalities for complete remission (CR) rates and the overall survival (OS) in adult ALLs.Materials and Methods: Conventional cytogenetics and fluorescence in situ hybridization were performed on bone marrow cells from 33 newly-diagnosed ALL adults. Two karyotype categories [standard- risk group- normal karyotype, hyperdiplody and other structural aberrations, and high-risk group-t(11q23)/MLL, t(9;22)/bcr-abl, t(1;19), t(8;14), C-MYC and complex karyotype] and the biologically and clinically relevant ALL ploidy subgroups were prospectively defined.Results: Chromosomal abnormalities were found in 52% of the cases with a high rate of poor-risk translocations - t(9;22), t(8q24), t(11q23), t(1;19). The total CR rate was 67% and the median time for achievement 2.33 months. Male sex, an age below 35 years and the absence of high risk translocations might have contributed to the high CR rates. Female patients, hyperdiplody, low white blood cells (WBC), and random cytogenetic aberrations had the longest OS. OS, 3- and 5-years survival periods were significantly shorter for poor-risk than standard risk group (p=.015, p=.001 and p=.005, respectively).Conclusion: This study emphasizes the lack of influence of cytogenetic aberrations on the CR and the time to achieve CR. However, our observations show that these aberrations are an independent prognostic factor in adult ALL - they allow predicting therapy resistance and the OS time after intensetreatment

MIK2 is a candidate gene of the S-locus for sporophytic self-incompatibility in chicory (Cichorium intybus, Asteraceae)

The Cichorium genus offers a unique opportunity to study the sporophytic self-incompatibility (SSI) system, being composed of species characterized by highly efficient self-incompatibility (e.g., C. intybus) and complete self-compatibility (e.g., C. endivia). To this end, the chicory genome was used to map seven previously identified SSI locus-associated markers. The region containing the S-locus was therefore restricted to an similar to 4 M bp window on chromosome 5. Among the genes predicted in this region, MDIS1 INTERACTING RECEPTOR LIKE KINASE 2 (ciMIK2) was particularly promising as a candidate for SSI. Its ortholog in Arabidopsis (atMIK2) is involved in pollen-stigma recognition reactions, and its protein structure is similar to that of S-receptor kinase (SRK), a key component of the SSI system in the Brassica genus. The amplification and sequencing of MIK2 in chicory and endive accessions revealed two contrasting scenarios. In C. endivia, MIK2 was fully conserved even when comparing different botanical varieties (i.e., smooth and curly endive). In C. intybus, 387 polymorphic positions and 3 INDELs were identified when comparing accessions of different biotypes all belonging to the same botanical variety (i.e., radicchio). The polymorphism distribution throughout the gene was uneven, with hypervariable domains preferentially localized in the LRR-rich extracellular region, putatively identified as the receptor domain. The gene was hypothesized to be under positive selection, as the nonsynonymous mutations were more than double the synonymous ones (dN/dS = 2.17). An analogous situation was observed when analyzing the first 500 bp of the MIK2 promoter: no SNPs were observed among the endive samples, whereas 44 SNPs and 6 INDELs were detected among the chicory samples. Further analyses are needed to confirm the role of MIK2 in SSI and to demonstrate whether the 23 species-specific nonsynonymous SNPs in the CDS and/or the species-specific 10 bp-INDEL found in a CCAAT box region of the promoter are responsible for the contrasting sexual behaviors of chicory and endive

Reducing the polynomial-like iterative equations order and a generalized Zoltan Boros' problem

We present a technique for reducing the order of polynomial-like iterative equations; in particular, we answer a question asked by Wenmeng Zhang and Weinian Zhang. Our method involves the asymptotic behaviour of the sequence of consecutive iterates of the unknown function at a given point. As an application we solve a generalized problem of Zoltán Boros posed during the 50th ISFE

Long-term Maintenance of Reduced Intraocular Pressure by Daily or Twice Daily Topical Application of Prostaglandins to Cat or Rhesus Monkey Eyes

Substantial evidence indicates that a single topical application of prostaglandins (PGs) can reduce intraocular pressure (IOP) in the eyes of several species. However, earlier literature, dealing with ocular hypertensive and inflammatory responses, shows the development of tachyphylaxis to subsequent doses of PGs. If similar tolerance developed to the ocular hypotensive effects of PGs, it would preclude the use of these agents in the treatment of chronic glaucoma. The present study shows, however, that although tachyphylaxis to the ocular hypotensive effects of PGs develops in rabbits, this is not a typical response among mammals. Significant IOP reduction was maintained in cats for up to 9 months by topical application of PGE 2 at 12-, 24-, or 48-hr intervals. The IOP reduction was jeopardized seriously only when the PG was applied every other day for several days or when, on a few occasions, 3 days were allowed to elapse between PGE 2 applications. Ocular hypotension was also maintained during the course of topical treatment of rhesus monkey eyes with PGF 2a . Short periods of pupillary constriction followed the application of each dose of PGF 2a to cat eyes, but the miotic response of rhesus monkeys to PGF 2a and cats to PGE 2 was negligible. Other apparent side effects were noted, but none of these were severe or progressive. These results clearly demonstrate that tachyphylaxis, or tolerance, is not expected to present an obstacle to the development of eicosanoids and/or their derivatives as therapeutic agents for the long-term treatment of ocular hypertension and chronic glaucoma. Invest Ophthalmol Vis Sci 24: [312][313][314][315][316][317][318][319] 1983 Early studies on the effects of prostaglandins (PGs) on the eye, primarily designed to determine the role of these autacoids in the ocular irritative response, concluded that exogenous PGs can produce increased intraocular pressure (IOP) and breakdown of the blood-aqueous barrier. 1 It has recently been shown, however, that appropriate doses of PGF 2a topically applied to the eyes of rabbits, 2 owl monkeys, 3 rhesus monkeys, and cats 4 can, in fact, reduce rather than increase IOP. In the rabbit there is only a narrow margin between the hypotensive dose of PGF 2a and a dose that causes initial hypertension. 2 Ten-to 100-fold higher PGF 2a doses than those that reduce IOP in the rabbit are required to produce the same effect in owl monkeys, rhesus monkeys, and cats, but eve

Splenectomy through the laparoscopic approach and how I do it

Scopul lucrării. Studierea cazurilor de splenectomie prin abord laparoscopic și clasic în vederea stabilirii cauzalității tip procedurăincidente perioperatorii și demonstrării superiorității abordului laparoscopic. Materiale și metode. Au fost selectate cazurile de splenectomie efectuate prin abord clasic și laparoscopic în Clinica de Chirurgie Colțea, perioada 01.01.2019-31.12.2022. Am revăzut filmările intraoperatorii, am evaluat factorii generali și locali asociați abordului chirurgical preferat. Rezultate. Au fost selectate 29 de cazuri, vârsta medie 54.8 ani, 72.4% femei, 27.6% barbati, 55.2% (16) cu multiple comorbidități: antecedente chirurgicale majore, insuficiența cardiacă, asmul bronșic sever etc. Cazurile au fost împărțite în 2 grupe: Gr. I- cu abord laparoscopic 34.5% (10), Gr. II- cu abord clasic 65.5% (19). Incidența comorbidităților a fost de 10% (1) în Gr.I, cu dimensiunea medie a splinei de 13.2 cm (min 8.5 cm, max 21 cm) și 84.2 % (16) în Gr.II cu dimensiunea medie a splinei de 20.4 cm (min 10 cm, max 34 cm). Doar în Gr.II au fost înregistrate sângerări intraoperatorii în 78.9% (15), iar in 15.8% (3) au fost complicații postoperatorii. Pacienții din Gr.I au avut o perioadă de spitalizare postoperatorie medie de 4 zile, iar cei din Gr. II de 7 zile și au fost externați cu o evoluție postoperatorie favorabilă. Concluzie. Abordul laparoscopic are aceleași indicații ca abordul clasic conform EAES, cu avantajul complicațiilor perioperatorii mult reduse și o spitalizare postoperatorie mai mică comparativ cu abordul clasic, iar în cazul echipelor experimentate este posibil abordul laparoscopic inclusiv la pacienții cu splenomegalii masive (>20cm), care poate fi asistat manual.Aim of study. Study of cases of splenectomy by laparoscopic and classic approach in order to establish causality type procedureperioperative incidents and demonstrate the superiority of the laparoscopic approach. Materials and methods. The cases of splenectomy performed by classical and laparoscopic approach in the Colțea Surgery Clinic, period 01.01.2019-31.12.2022, were selected. We reviewed intraoperative films, assessed general and local factors associated with the preferred surgical approach. Results. 29 cases were selected, average age 54.8 years, 72.4% women, 27.6% men, 55.2% (16) with multiple comorbidities: major surgical antecedents, heart failure, severe bronchial asthma, etc. The cases were divided into 2 groups: Gr. I- with laparoscopic approach 34.5% (10), Gr. II- with classic approach 65.5% (19). The incidence of comorbidities was 10% (1) in Gr.I, with mean spleen size of 13.2 cm (min 8.5 cm, max 21 cm) and 84.2% (16) in Gr.II with mean spleen size of 20.4 cm (min 10 cm, max 34 cm). Only in Gr.II, intraoperative bleeding was recorded in 78.9% (15), and in 15.8% (3) there were postoperative complications. The patients in Gr. I had an average postoperative hospitalization period of 4 days, and those in Gr. II of 7 days, and were discharged with a favorable postoperative evolution. Conclusions. The laparoscopic approach has the same indications as the classic one according to EAES, with the advantage of reduced perioperative complications and a shorter postoperative hospitalization, and in the case of experienced teams, the laparoscopic approach is possible, including patients with massive splenomegaly (>20cm), being manually assisted

Nanoscale control of Ag nanostructures for plasmonic fluorescence enhancement of near-infrared dyes

Potential utilization of proteins for early detection and diagnosis of various diseases has drawn considerable interest in the development of protein-based detection techniques. Metal induced fluorescence enhancement offers the possibility of increasing the sensitivity of protein detection in clinical applications. We report the use of tunable plasmonic silver nanostructures for the fluorescence enhancement of a near-infrared (NIR) dye (Alexa Fluor 790). Extensive fluorescence enhancement of ∼2 orders of magnitude is obtained by the nanoscale control of the Ag nanostructure dimensions and interparticle distance. These Ag nanostructures also enhanced fluorescence from a dye with very high quantum yield (7.8 fold for Alexa Fluor 488, quantum efficiency (Qy) = 0.92). A combination of greatly enhanced excitation and an increased radiative decay rate, leading to an associated enhancement of the quantum efficiency leads to the large enhancement. These results show the potential of Ag nanostructures as metal induced fluorescence enhancement (MIFE) substrates for dyes in the NIR “biological window” as well as the visible region. Ag nanostructured arrays fabricated by colloidal lithography thus show great potential for NIR dye-based biosensing applications

Maternal complications following open and fetoscopic fetal surgery: A systematic review and meta-analysis

OBJECTIVE: To establish maternal complication rates for fetoscopic or open fetal surgery. METHODS: We conducted a systematic literature review for studies of fetoscopic or open fetal surgery performed since 1990, recording maternal complications during fetal surgery, the remainder of pregnancy, delivery and after the index pregnancy. RESULTS: One hundred and sixty-six studies were included, reporting outcomes for open fetal (n=1193 patients) and fetoscopic surgery (n=9403 patients). No maternal deaths were reported. The risk of any maternal complication in the index pregnancy was 20.9% (95%CI 15.22-27.13) for open fetal and 6.2% (95%CI 4.93-7.49) for fetoscopic surgery. For severe maternal complications (Grade III to V Clavien-Dindo classification of surgical complications) the risk was 4.5% (95%CI 3.24-5.98) for open fetal and 1.7% (95%CI 1.19-2.20) for fetoscopic surgery. In subsequent pregnancies, open fetal surgery increased the risk of preterm birth but not uterine dehiscence or rupture. Nearly one quarter of reviewed studies (n=175, 23.3%) were excluded for failing to report the presence or absence of maternal complications. CONCLUSIONS: Maternal complications occur in 6.2% fetoscopic and 20.9% open fetal surgeries, with serious maternal complications in 1.7% fetoscopic and 4.5% open procedures. Reporting of maternal complications is variable. To properly quantify maternal risks, outcomes should be reported consistently across all fetal surgery studies