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Noa Alon,1,3,* Yana Miroshnikov,2,3,* Nina Perkas,2,3 Ifat Nissan,2,3 Aharon Gedanken,2,3 Orit Shefi1,31Faculty of Engineering, 2Department of Chemistry, 3Bar-Ilan Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat Gan, Israel*These authors contributed equally to this workAbstract: Much effort has been devoted to the design of effective biomaterials for nerve regeneration. Here, we report the novel use of silver nanoparticles (AgNPs) as regenerative agents to promote neuronal growth. We grew neuroblastoma cells on surfaces coated with AgNPs and studied the effect on the development of the neurites during the initiation and the elongation growth phases. We find that the AgNPs function as favorable anchoring sites, and the growth on the AgNP-coated substrates leads to a significantly enhanced neurite outgrowth. Cells grown on substrates coated with AgNPs have initiated three times more neurites than cells grown on uncoated substrates, and two times more than cells grown on substrates sputtered with a plain homogenous layer of silver. The growth of neurites on AgNPs in the elongation phase was enhanced as well. A comparison with substrates coated with gold nanoparticles (AuNPs) and zinc oxide nanoparticles (ZnONPs) demonstrated a clear silver material-driven promoting effect, in addition to the nanotopography. The growth on substrates coated with AgNPs has led to a significantly higher number of initiating neurites when compared to substrates coated with AuNPs or ZnONPs. All nanoparticle-coated substrates affected and promoted the elongation of neurites, with a significant positive maximal effect for the AgNPs. Our results, combined with the well-known antibacterial effect of AgNPs, suggest the use of AgNPs as an attractive nanomaterial – with dual activity – for neuronal repair studies.Keywords: nerve regeneration, nanotopography, antibacterial material, neuroblastoma, gold nanoparticles, zinc oxide nanoparticles

Abstract We argue that on-shell excitations with large negative energies are created rapidly when the string coupling increases with time. This does not indicate an inconsistency in string theory since the negative energy on-shell excitation is always entangled with an on-shell excitation with a positive energy. The total energy of this energy-EPR state vanishes. We discuss the reason the energy-EPR states appear in string theory and the role they might play in black hole physics.

Abstract We point out that in many eternal black holes, including a Schwarzschild eternal black hole and an eternal black hole in AdS 5, instant folded strings are created in the past wedge and render the region just outside the horizon singular. We also make a conjecture regarding instant folded D-branes and discuss their possible implications for eternal black holes. In particular, we argue that the bulk modes responsible for Poincare recurrence, when it occurs in the dual quantum field theory, are either instant folded strings or instant folded D-branes.

Abstract Time-like linear dilaton backgrounds admit a classical solution that describes a closed folded string that is created at an instant. We refer to such strings as Instant Folded Strings (IFS). We study an exact worldsheet CFT description of an IFS that involves two vertex operators which describe two open string modes that propagate on a time-like FZZT-brane, which plays the role of a regulator to the IFS. We take advantage of this description to calculate the most basic quantity associated with IFSs — their production rate. Some implications of this calculation to stringy cosmology and black hole interior are briefly discussed.

Abstract We consider the backreaction of the winding condensate on the cigar background. We focus on the case of the SL(2, ℝ) k /U(1) cigar associated with, e.g., the near-horizon limit of k NS5 black-branes. We solve the equations of motion numerically in the large k limit as a function of the amplitude, A, of the winding mode at infinity. We find that there is a critical amplitude, A c = exp(−γ/2), that admits a critical solution. In string theory, the exact SL(2, ℝ) k /U(1) cigar CFT fixes completely the winding amplitude, A s , at infinity. We find that in the large k limit there is an exact agreement, A c = A s . The critical solution is a cigar with a puncture at its tip; consequently, the black-hole entropy is carried entirely by the winding condensate. We argue that, in the Lorentzian case, the information escapes the black hole through this puncture.

Abstract Time-like linear dilaton triggers, at the classical level, the creation of closed folded strings at an instant. We show that in cosmology these instant folded strings induce negative pressure at no energy cost. Hence they seem to allow an era in which the energy density increases (decreases) while the universe is expanding (contracting). This and other aspects of instant folded strings suggest that they might shed new light on the origin of the arrow of time.

Abstract We study the recent claim that black NS5-branes are filled with folded strings. We calculate, in the near-extremal case, the number of folded strings at the black fivebranes interior, using different approaches, and get the exact same answer. The backreaction of the folded strings leads us to argue that the interior of the black fivebrane is AdS 2 (times a compact manifold) and that infalling matter cannot reach the interior, due to a shock wave at the horizon. These considerations also suggest a novel insight into the black fivebranes entropy.

Abstract We show that in string theory, due to non-perturbative effects, there are cases in which two states that semi-classically are completely different, are in fact the same. One state cannot be excited without exciting the other; they are two components of the same state in the exact theory. As a result, in some situations that include black holes, the nature of information in string theory is dramatically different than in field theory. In particular, each general-relativity state, that lives in the atmosphere of black fivebranes, is accompanied with an excitation that lives on folded strings, which fill the black-hole interior. This is likely related to the way that information is extracted from black holes in string theory, and we refer to it as stringy information.

Abstract It is well known that non-perturbative α ′ corrections to the SL(2, ℝ)/U(1) cigar geometry are described via a condensation of a Sine-Liouville operator that schematically can be written as W + + W − , where W ± describe a string with winding number ±1. This condensation leads to interesting effects in the cigar geometry that take place already at the classical level in string theory. Condensation of the analytically continued Sine-Liouville operator in the Lorentzian SL(2, ℝ)/U(1) black hole is problematic. Here, we propose that in the black hole case, the non-perturbative α ′ corrections are described in terms of an operator that can be viewed as the analytic continuation of the fusion of W + and W − . We show that this operator does not suffer from the same problem as the analytically continued Sine-Liouville operator and argue that it describes folded strings that fill the entire black hole and, in a sense, replace the black hole interior. We estimate the folded strings radiation, and show that they radiate at the Hawking temperature.

We continue our study of string theory in a background that interpolates between AdS3 in the infrared and a linear dilaton spacetime IR1,1×IRϕ in the UV. This background corresponds via holography to a CFT2 deformed by an operator of dimension (2,2). We discuss the structure of spatial entanglement in this model, and compare it to the closely related TT‾ deformed CFT2.

Abstract We show that negative (∇Φ)2, where Φ is the dilaton, leads to a rapid creation of folded strings. Consequently it appears that the interior of the SL(2, ℝ) k /U(1) black hole is not empty, but is filled with folded strings.

We continue our study of string theory in a background that interpolates between AdS3 in the infrared and a linear dilaton spacetime R1,1×Rϕ in the UV. This background corresponds via holography to a CFT2 deformed by a certain irrelevant operator of dimension (2,2). We show that for two point functions of local operators in the infrared CFT, conformal perturbation theory in this irrelevant operator has a finite radius of convergence in momentum space, and one can use it to flow up the renormalization group. The spectral density develops an imaginary part above a certain critical value of the spectral parameter; this appears to be related to the non-locality of the theory. In position space, conformal perturbation theory has a vanishing radius of convergence; the leading non-perturbative effect is an imaginary part of the two point function.

Abstract We elaborate on the recent claim [1] that non-perturbative effects in α′, which are at the core of the FZZ duality, render the region just behind the horizon of the SL(2, ℝ) k /U(1) black hole singular already at the classical level (g s = 0). We argue that the 2D classical SL(2, ℝ) k /U(1) black hole could shed some light on quantum black holes in higher dimensions including large black holes in AdS5 × S 5.

Abstract Recently we proposed a universal solvable irrelevant deformation of AdS3 /CFT2 duality, which leads in the ultraviolet to a theory with a Hagedorn entropy [1]. In this note we provide a worldsheet description of this theory as a coset CFT, and compare its spectrum to the field theory predictions of [2, 3].

Abstract It was recently shown that the theory obtained by deforming a general two dimensional conformal theory by the irrelevant operator T T ¯ $$ T\overline{T} $$ is solvable. In the context of holography, a large class of such theories can be obtained by studying string theory on AdS 3. We show that a certain single trace T T ¯ $$ T\overline{T} $$ deformation of the boundary CFT 2 gives rise in the bulk to string theory in a background that interpolates between AdS 3 in the IR and a linear dilaton spacetime in the UV, i.e. to a two dimensional vacuum of Little String Theory. This construction provides holographic duals for a large class of vacua of string theory in asymptotically linear dilaton spacetimes, and sheds light on the UV behavior of T T ¯ $$ T\overline{T} $$ deformed CFT 2. It may provide a step towards holography in flat spacetime.

Abstract The potential behind the horizon of an eternal black hole in classical theories is described in terms of data that is available to an external observer — the reflection coefficient of a wave that scatters on the black hole. In GR and perturbative string theory (in α′), the potential is regular at the horizon and it blows up at the singularity. The exact reflection coefficient, that is known for the SL(2, ℝ) k /U(1) black hole and includes non-perturbative α′ effects, seems however to imply that there is a highly non-trivial structure just behind the horizon.

Recently it was shown that folded strings are spontaneously created behind the horizon of the SL(2,R)k/U(1) black hole. Here we show that these folded strings violate the averaged null energy condition macroscopically. We discuss possible consequences of this observation on black hole physics in general, and on the information puzzle in particular.

Coinfection of HIV and multidrug-resistant tuberculosis (MDR-TB) presents significant challenges in terms of the treatment and prognosis of tuberculosis, leading to complexities in managing the disease and impacting the overall outcome for TB patients. This study presents a remarkable case of a patient with MDR-TB and HIV coinfection who survived for over 8 years, despite poor treatment adherence and comorbidities. Whole genome sequencing (WGS) of the infecting Mycobacterium tuberculosis ( Mtb ) strain revealed a unique genomic deletion, spanning 18 genes, including key genes involved in hypoxia response, intracellular survival, immunodominant antigens, and dormancy. This deletion, that we have called "Del-X," potentially exerts a profound influence on the bacterial physiology and its virulence. Only few similar deletions were detected in other non-related Mtb genomes worldwide. In vivo evolution analysis identified drug resistance and metabolic adaptation mutations and their temporal dynamics during the patient's treatment course., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Rubinstein, Makhon, Losev, Valenci, Gatt, Margalit, Fass, Kutikov, Murik, Zeevi, Savyon, Tau, Kaidar Shwartz, Dveyrin, Rorman and Nissan.)

According to World Health Organization WHO, Tuberculosis (TB) is the second cause of death from infectious disease worldwide. During 2021, 10.6 million people were infected with TB, and 1.6 million people died. TB is caused by pathogens belonging to the Mycobacterium tuberculosis complex (MTBC), mainly by Mycobacterium tuberculosis ( M.tb ). Members of this complex are acid-fast bacilli, which can cause intrapulmonary and extra pulmonary TB, and can be divided into various lineages, based on genomic markers. The main public health threat comes from drug resistant M.tb strains, which are responsible for about 25% of TB death and treatment failure worldwide. Treating drug resistant TB patients significantly raises the costs of TB treatment. This study provides an overview of the demographic and drug susceptibility characteristics of newly diagnosed TB patients in Israel in 2021. The State of Israel has a very low level of TB endemicity and is at a pre-elimination phase. Notably, only 11.7% of the newly diagnosed TB patients were born in Israel. In this report, of the 154 new laboratory-confirmed TB patients, 66.7% had pulmonary TB, while 16% had extrapulmonary TB. Males accounted for 52% of the patients, with the most prevalent age group being 21-40. Most patients were citizens of Israel (53.9%), while 37.7% had no Israeli citizenship. Among non-citizens, there was a predominance of males and patients aged 21-40. The susceptibility profile showed a high resistance rate to streptomycin (18.2%) and to a lower extent to isoniazid (13.6%), pyrazinamide (8.4%), rifampicin (7.8%), and ethambutol (3.2%). Only 2 cases of XDR-TB and 10 MDR-TB strains were detected in Israel in 2021, with both XDR strains and 5 out of 10 MDR strains belonging to the Beijing lineage. Most of Beijing isolates were resistant to at least one tested drug. Genomic sequencing of 134 out of 156 strains and bioinformatics analysis using the MTBseq program and WHO mutation catalogue shows a good match with only 9 discrepancies between phenotypic and genotypic susceptibility profiles in first line drugs. The most common lineage is Delhi-Cas (23%) followed by the Beijing lineage (17%). Most patients from the Delhi-Cas lineage were born in Africa, while patients with Beijing isolates were born in different countries. Minimum spanning tree analysis identified 15 clusters. The study highlights the need for ongoing surveillance of TB using molecular and phenotypic tools to further decreasing the spreading level of the disease and develop effective treatment strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Losev, Rubinstein, Nissan, Haviv, Barsky, Volinsky, Bar-Giora, Zouher, Hamawi, Valenci, Kutikov, Shwartz, Dveyrin, Chemtob and Rorman.)

Introduction: GBS may cause a devastating disease in newborns. In early onset disease of the newborn the bacteria are acquired from the colonized mother during delivery. We characterized type VII secretion system (T7SS), exporting small proteins of the WXG100 superfamily, in group B Streptococci (GBS) isolates from pregnant colonized women and newborns with early onset disease (EOD) to better understand T7SS contribution to virulence in these different clinical scenarios., Methods: GBS genomes [N=33, 17 EOD isolates (serotype III/ST17) and 16 colonizing isolates (12 serotype VI/ST1, one serotype VI/ST19, one serotype VI/ST6, and two serotype 3/ST19)] were analyzed for presence of T7SS genes and genes encoding WXG100 proteins. We also perform bioinformatic analysis. Galleria mellonella larvae were used to compare virulence between colonizing, EOD, and mutant EOD isolates. The EOD isolate number 118659 (III/ST17) was used for knocking out the essC gene encoding a membrane-bound ATPase, considered the driver of T7SS., Results: Most GBS T7SS loci encoded core component genes: essC, membrane-embedded proteins (essA; essB), modulators of T7SS activity (esaA; esaB; esaC) and effectors: [esxA (SAG1039); esxB (SAG1030)].Bioinformatic analysis indicated that based on sequence type (ST) the clinicalGBS isolates encode at least three distinct subtypes of T7SS machinery. In all ST1isolates we identified two copies of esxA gene (encoding putative WXG100proteins), when only 23.5% of the ST17 isolates harbored the esxA gene. Five ST17isolates encoded two copies of the essC gene. Orphaned WXG100 molecule(SAG0230), distinct from T7SS locus, were found in all tested strains, except inST17 strains where the locus was found in only 23.5% of the isolates. In ST6 andST19 isolates most of the structure T7SS genes were missing. EOD isolates demonstrated enhanced virulence in G. mellonella modelcompared to colonizing isolates. The 118659DessC strain was attenuated in itskilling ability, and the larvae were more effective in eradicating 118659DessC., Conclusions: We demonstrated that T7SS plays a role during infection. Knocking out the essC gene, considered the driver of T7SS, decreased the virulence of ST17 responsible for EOD, causing them to be less virulent comparable to the virulence observed in colonizing isolates., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Schindler, Rahav, Nissan, Valenci, Ravins, Hanski, Ment, Tekes-Manova and Maor.)

Perihepatitis (Fitz-Hugh-Curtis syndrome) is a rare complication of sexually transmitted infections, mostly seen in women. Only 12 male cases have been reported to date, of which Chlamydia trachomatis was confirmed in 2. We report a case of chlamydial perihepatitis in a male patient, occurring 1 month after Mpox and associated with the unusual LGV ST23 strain. Our case suggests that rectal Mpox lesions may facilitate chlamydial dissemination., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Group B streptococcus (GBS) harbors many virulence factors but there is limited data regarding their importance in colonization in pregnancy and early-onset disease (EOD) in the newborn. We hypothesized that colonization and EOD are associated with different distribution and expression of virulence factors., Methods: We studied 36 GBS EOD and 234 GBS isolates collected during routine screening. Virulence genes (pilus-like structures- PI-1, PI-2a, PI-2b ; rib and hvgA ) presence and expression were identified by PCR and qRT-PCR. Whole genome sequencing (WGS) and comparative genomic analyses were used to compare coding sequences (CDSs) of colonizing and EOD isolates., Results: Serotype III (ST17) was significantly associated with EOD and serotype VI (ST1) with colonization. hvgA and rib genes were more prevalent among EOD isolates (58.3 and 77.8%, respectively; p < 0.01). The pilus loci PI-2b and PI-2a were more prevalent among EOD isolates (61.1%, p < 0.01), while the pilus loci PI-2a and PI-1 among colonizing isolates (89.7 and 93.1% vs. 55.6 and 69.4%, p < 0.01). qRT PCR analysis revealed that hvgA was barely expressed in colonizing isolates, even though the gene was detected. Expression of the rib gene and PI-2b was two-fold higher in EOD isolates compared to colonizing isolates. Transcription of PI-2a was three-fold higher in colonizing isolates compared to EOD isolates. ST17 isolates (associated with EOD) had a smaller genome size compared ST1 and the genome was more conserved relative to the reference strain and ST17 isolates. In a multivariate logistic regression analysis virulence factors independently associated with EOD were serotype 3, and PI-1 and PI-2a was protective., Conclusion: There was a significant difference in the distribution of hvg A , rib , and PI genes among EOD (serotype III/ST17) and colonizing (serotype VI/ST1) isolates suggesting an association between invasive disease and these virulence factors. Further study is needed to understand the contribution of these genes to GBS virulence., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Schindler, Rahav, Nissan, Treygerman, Prajgrod, Attia, Raz, Valenci, Tekes-Manova and Maor.)

Background: antimicrobial resistance is a global problem in human and veterinary medicine. We aimed to investigate the extended spectrum β-lactamase-producing Enterobacterales (ESBL-PE) gut colonization in healthy community dogs in Israel., Methods: Rectal swabs were sampled from 145 healthy dogs, enriched, plated on selective plates, sub-cultured to obtain pure cultures, and ESBL production was confirmed. Bacterial species and antibiotic susceptibility profiles were identified. WGS was performed on all of the ESBL-PE isolates and their resistomes were identified in silico. Owners' questionnaires were collected for risk factor analysis., Results: ESBL-PE gut colonization rate was 6.2% ( n = 9/145, 95% CI 2.9-11.5). Overall, ten isolates were detected (one dog had two isolates); the main species was Escherichia coli (eight isolates), belonging to diverse phylogenetic groups-B1, A and C. Two isolates were identified as Citrobacter braakii , and C. portucalensis . A phylogenetic analysis indicated that all of the isolates were genetically unrelated and sporadic. The isolates possessed diverse ESBL genes and antibiotic-resistance gene content, suggesting independent ESBL spread. In a multivariable risk factor analysis, coprophagia was identified as a risk factor for ESBL-PE gut colonization ( p = 0.048, aOR = 4.408, 95% CI 1.014-19.169)., Conclusions: healthy community dogs may be colonized with ESBL-PE MDR strains, some of which were previously reported in humans, that carry wide and diverse resistomes and may serve as a possible source for AMR.

Transmission of reductive and oxidative cues from the photosynthetic electron transport chain to redox regulatory protein networks plays a crucial role in coordinating photosynthetic activities. The tight balance between these two signals dictates the cellular response to changing light conditions. While the role of reductive signals in activating chloroplast metabolism is well established, the role of their counterbalanced oxidative signals is still unclear, mainly due to monitoring difficulties. Here, we introduced chl-roGFP2-PrxΔCR, a 2-Cys peroxiredoxin-based biosensor, into Arabidopsis thaliana chloroplasts to monitor the dynamic changes in photosynthetically derived oxidative signaling. We showed that chl-roGFP2-PrxΔCR oxidation states reflected oxidation patterns similar to those of endogenous 2-Cys peroxiredoxin under varying light conditions. By employing a set of genetically encoded biosensors, we showed the induction of 2-Cys peroxiredoxin-dependent oxidative signals, throughout the day, under varying light intensities and their inverse relationship with NADPH levels, unraveling the combined activity of reducing and oxidizing signals. Furthermore, we demonstrated the induction of 2-Cys peroxiredoxin-derived oxidative signals during a dark–to–low-light transition and uncovered a faster increase in carbon assimilation rates during the photosynthesis induction phase in plants deficient in 2-Cys peroxiredoxins compared with wild type, suggesting the involvement of oxidative signals in attenuating photosynthesis. The presented data highlight the role of oxidative signals under nonstress conditions and suggest that oxidative signals measured by peroxiredoxin-based biosensors reflect the limitation to photosynthesis imposed by the redox regulatory system.

In 2018, Mycobacterium canariasense bloodstream infection was diagnosed in Israel. Further investigation had identified additional five cases in three medical centers, including isolates from blood (1), cornea (1), and sputum (3). Isolates were susceptible to all the antimicrobial tested. All but one isolate was related by whole-genome phylogeny., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections are common among men who have sex with men (MSM). Many oropharyngeal and anorectal infections remain asymptomatic. We aimed to evaluate triple-site screening following PrEP introduction. We enrolled a prospective cohort study including 210 asymptomatic MSM during 2019-2020, analyzed by groups: HIV positive (HIV+), HIV-uninfected using PrEP (HIV-/PrEP+), or HIV-uninfected not using PrEP (HIV-/PrEP-). A self-administered questionnaire captured demographic information and sexual risk-taking behaviors. CT/NG testing results were compared between study groups and predictors of infection were evaluated. We included 59 HIV+, 70 HIV-/PrEP+, and 81 HIV-/PrEP- subjects. 30% ( n = 62) of participants tested positive for CT/NG. HIV-/PrEP+ group had highest proportion of infections ( n = 33, 47%) followed by HIV-/PrEP- ( n = 16, 22%) and HIV+ ( n =13, 20%; p < .001). Importantly, 98% (80/82) of pharyngeal/anorectal CT/NG infections were missed in genitourinary tract screening alone. PrEP use and previous syphilis infection were the strongest risk factor for CT/NG. Extra-genital asymptomatic CT/NG infections were prevalent among MSM. These data highlight the importance of routine extra-genital CT/NG testing in asymptomatic sexually active MSM. The study describes the consequences for three-site testing lack of implementation in the PrEP era.

The changing nature of the SARS-CoV-2 pandemic poses unprecedented challenges to the world's health systems. Emerging spike gene variants jeopardize global efforts to produce immunity and reduce morbidity and mortality. These challenges require effective real-time genomic surveillance solutions that the medical community can quickly adopt. The SARS-CoV-2 spike protein mediates host receptor recognition and entry into the cell and is susceptible to generation of variants with increased transmissibility and pathogenicity. The spike protein is the primary target of neutralizing antibodies in COVID-19 patients and the most common antigen for induction of effective vaccine immunity. Tight monitoring of spike protein gene variants is key to mitigating COVID-19 spread and generation of vaccine escape mutants. Currently, SARS-CoV-2 sequencing methods are labor intensive and expensive. When sequence demands are high sequencing resources are quickly exhausted. Consequently, most SARS-CoV-2 strains are sequenced in only a few developed countries and rarely in developing regions. This poses the risk that undetected, dangerous variants will emerge. In this work, we present HiSpike, a method for high-throughput cost effective targeted next generation sequencing of the spike gene. This simple three-step method can be completed in < 30 h, can sequence 10-fold more samples compared to conventional methods and at a fraction of their cost. HiSpike has been validated in Israel, and has identified multiple spike variants from real-time field samples including Alpha, Beta, Delta and the emerging Omicron variants. HiSpike provides affordable sequencing options to help laboratories conserve resources for widespread high-throughput, near real-time monitoring of spike gene variants., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Fass, Zizelski Valenci, Rubinstein, Freidlin, Rosencwaig, Kutikov, Werner, Ben-Tovim, Bucris, Erster, Zuckerman, Mor, Mendelson, Dveyrin, Rorman and Nissan.)

Non-tuberculous mycobacteria (NTM) are opportunistic pathogens that cause illness primarily in the elderly, in the immunocompromised or in patients with underlying lung disease. Since 2013, a global outbreak of NTM infection related to heater-cooler units (HCU) used in cardio-thoracic surgery has been identified. This outbreak was caused by a single strain of Mycobacterium intracellulare subsp. chimaera . In order to estimate the prevalence of this outbreak strain in Israel, we sampled Mycobacterium intracellulare subsp. chimaera from several HCU machines in Israel, as well as from patients, sequenced their genomes and compared them to the outbreak strain. The presence of mixed mycobacteria species in the samples complicated the analysis of obtained sequences. By applying a metagenomic binning strategy, we were able to obtain, and characterize, genomes of single strains from the mixed samples. Mycobacterium intracellulare subsp. chimaera strains were compared to each other and to previously reported genomes from other countries. The strain causing the outbreak related to the HCU machines was identified in several such machines in Israel but not in any clinical sample.

Previously, we identified a Chlamydia trachomatis lymphogranuloma venereum (LGV) recombinant strain possessing a non-LGV ompA genotype. Here, culture-independent genome sequencing confirms its circulation in Europe, Middle East, and North America, and unveils emergence of antibiotic resistance. Broad surveillance is needed., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Background: Several Multilocus Sequence Typing (MLST) schemes have been developed for Chlamydia trachomatis. Bom's MLST scheme for MLST is based on nested PCR amplification and sequencing of five hypervariable genes and ompA. In contrast to other Chlamydia MLST schemes, Bom's MLST scheme gives higher resolution and phylogenetic trees that are comparable to those from whole genome sequencing. However, poor results have been obtained with Bom's MLST scheme in clinical samples with low concentrations of Chlamydia DNA., Results: In this work, we present an improved version of the scheme that is based on the same genes and MLST database as Bom's MLST scheme, but with newly designed primers for nested-1 and nested-2 steps under stringent conditions. Furthermore, we introduce a third primer set for the sequencing step, which considerably improves the performance of the assay. The improved primers were tested in-silico using a dataset of 141 Whole Genome Sequences (WGS) and in a comparative analysis of 32 clinical samples. Based on cycle threshold and melting curve analysis values obtained during Real-Time PCR of nested-1 & 2 steps, we developed a simple scoring scheme and flow chart that allow identification of reaction inhibitors as well as to predict with high accuracy amplification success. The improved MLST version was used to obtain a genovars distribution in patients attending an STI clinic in Tel Aviv., Conclusions: The newly developed MLST version showed great improvement of assay results for samples with very low concentrations of Chlamydia DNA. A similar concept could be applicable to other MLST schemes.

Objectives: To study Group B Streptococcus (GBS) isolates associated with different clinical syndromes: asymptomatic carriage in pregnant women, intrauterine fetal death (IUFD), and early onset disease (EOD) in the newborn., Methods: GBS isolates were collected from asymptomatic pregnant women admitted for labor, IUFD cases, and neonates with EOD. Serotypes and antibiotic susceptibilities were determined. Multilocus sequence typing (MLST) was performed to assess genetic epidemiology., Results: GBS carriage rate was 26.1% (280/1074). The dominant serotype among asymptomatic pregnant women was VI [98/240 women (40.8%)], followed by serotypes III, V and IV in 42/240 (17.5%), 30/240 (12.5%) and 28/240 (11.7%) women, respectively. The dominant serotype in IUFD cases was serotype VI [10/13 (76.9%)]. In contrast the prevalent serotype among EOD cases was III [16/19 (84.2%)]. ST-1 was associated with IUFD [7/13 (53.8%)], ST-17 was associated with serotype III and EOD in the newborn 14/19 (73.7%)]. Erythromycin and clindamycin resistance reached 36.8%, 7.7% and 20.0%among EOD, vaginal carriage and IUFD, respectively., Conclusions: Serotypes VI and ST-1 were dominant among asymptomatic pregnant women and in IUFD cases while EOD was associated with serotype III and ST-17. Invasive mechanisms thus may differ between IUFD and EOD in the newborn and virulence may be related to capsule serotype. Resistance rates to erythromycin and clindamycin were high in EOD cases., Competing Interests: The authors have read the journal’s policy and the authors of this paper declare the following competing interests: YM and GR received consultant and lecture fees from Pfizer, MSD, Gilead and Astellas. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: We aimed to investigate the prevalence, molecular epidemiology and prevalence factors for Extended Spectrum β-Lactamase-producing Enterobacteriaceae (ESBL-E) shedding by race horses. A cross-sectional study was performed involving fecal samples collected from 169 Thoroughbred horses that were housed at a large racing facility in Ontario, Canada. Samples were enriched, plated on selective plates, sub-cultured to obtain pure cultures and ESBL production was confirmed. Bacterial species were identified and antibiotic susceptibility profiles were assessed. E. coli sequence types (ST) and ESBL genes were determined using multilocus sequence type (MLST) and sequencing. Whole genome sequencing was performed to isolates harboring CTX-M-1 gene. Medical records were reviewed and associations were investigated., Results: Adult horses (n = 169), originating from 16 different barns, were sampled. ESBL-E shedding rate was 12% (n = 21/169, 95% CI 8-18%); 22 ESBL-E isolates were molecularly studied (one horse had two isolates). The main species was E. coli (91%) and the major ESBL gene was CTX-M-1 (54.5%). Ten different E. coli STs were identified. Sixty-four percent of total isolates were defined as multi-drug resistant. ESBL-E shedding horses originated from 8/16 different barns; whereas 48% (10/21) of them originated from one specific barn. Overall, antibiotic treatment in the previous month was found as a prevalence factor for ESBL-E shedding (p = 0.016, prevalence OR = 27.72, 95% CI 1.845-416.555)., Conclusions: Our findings demonstrate the potential diverse reservoir of ESBL-E in Thoroughbred race horses. Multi-drug resistant bacteria should be further investigated to improve antibiotic treatment regimens and equine welfare.

The sequencing and bioinformatics analyses of isolates Cr150, Cr170, and Cr611 from powdered infant formula indicate that the three strains represent new members in the Cronobacter muytjensii , Cronobacter turicensis , and Cronobacter sakazakii groups, respectively., (Copyright © 2020 Zizelski Valenci et al.)

Low gibberellin (GA) activity in tomato (Solanum lycopersicum) inhibits leaf expansion and reduces stomatal conductance. This leads to lower transpiration and improved water status under transient drought conditions. Tomato has three GIBBERELLIN-INSENSITIVE DWARF1 (GID1) GA receptors with overlapping activities and high redundancy. We tested whether mutation in a single GID1 reduces transpiration without affecting growth and productivity. CRISPR-Cas9 gid1 mutants were able to maintain higher leaf water content under water-deficit conditions. Moreover, while gid1a exhibited normal growth, it showed reduced whole-plant transpiration and better recovery from dehydration. Mutation in GID1a inhibited xylem vessel proliferation, which led to lower hydraulic conductance. In stronger GA mutants, we also found reduced xylem vessel expansion. These results suggest that low GA activity affects transpiration by multiple mechanisms: it reduces leaf area, promotes stomatal closure, and reduces xylem proliferation and expansion, and as a result, xylem hydraulic conductance. We further examined if gid1a performs better than the control M82 in the field. Under these conditions, the high redundancy of GID1s was lost and gid1a plants were semi-dwarf, but their productivity was not affected. Although gid1a did not perform better under drought conditions in the field, it exhibited a higher harvest index., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Mastitis, inflammation of the mammary gland, is a common disease of dairy animals. The disease is caused by bacterial infection ascending through the teat canal and mammary pathogenic Escherichia coli (MPEC) are common etiology. In the first phase of infection, virulence mechanisms, designated as niche factors, enable MPEC bacteria to resist innate antimicrobial mechanisms, replicate in milk, and to colonize the mammary gland. Next, massive replication of colonizing bacteria culminates in a large biomass of microbe-associated molecular patterns (MAMPs) recognized by pattern recognition receptors (PRRs) such as toll-like receptors (TLRs) mediating inflammatory signaling in mammary alveolar epithelial cells (MAEs) and macrophages. Bacterial lipopolysaccharides (LPSs), the prototypical class of MAMPs are sufficient to elicit mammary inflammation mediated by TLR4 signaling and activation of nuclear factor kB (NF-kB), the master regulator of inflammation. Using in vivo mastitis model, in low and high complements mice, and in vitro NF-kB luminescence reporter system in MAEs, we have found that the smooth configuration of LPS O-polysaccharides in MPEC enables the colonizing organisms to evade the host immune response by reducing inflammatory response and conferring resistance to complement. Screening a collection of MPEC field strains, we also found that all strains were complement resistant and 94% (45/48) were smooth. These results indicate that the structure of LPS O-polysaccharides chain is important for the pathogenesis of MPEC mastitis and provides protection against complement-mediated killing. Furthermore, we demonstrate a role for complement, a key component of innate immunity, in host-microbe interactions of the mammary gland.