Vaccines against SARS-CoV-2 (COVID-19) proved beneficial for COVID-19 disease attenuation and preventing virus spreading. Cumulative reports of the rarity of antineutrophil cytoplasmic autoantibodies (ANCA)-associated vasculitis (AAV) raise concerns about its relationship with COVID-19 vaccination. Several case reports described ANCA-associated pauci-immune glomerulonephritis (ANCA-GN) following COVID-19 vaccination with some uniqueness. We systematically reviewed COVID-19 vaccine-induced ANCA-GN from PubMed, SCOPUS, and Cochrane library databases until 1 January 2023 according to PRISMA guidelines and presented our three cases. Twenty-six cases from 25 articles, including our 3 cases, were analyzed. Most cases were diagnosed following the second dose of the COVID-19 vaccine (59%) with a median (IQR) interval onset of 14 (16) days. The highest prevalence was related to the mRNA-type vaccine. Anti-myeloperoxidase (MPO) ANCA was far more common than the other ANCAs, with various positive autoantibodies. Fourteen cases  had extra-kidney AAV manifestation. Although severe kidney injury was observed in 10/29 (34%), remission was achieved in 89% (25/28) with no death. The mechanisms of the vaccine-inducing ANCA-GN were postulated here. Since ANCA-GN after the COVID-19 vaccine was rare, the benefit of the COVID-19 vaccine could outweigh the risk of ANCA-GN side effects in the pandemic era. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the ongoing global pandemic of coronavirus disease 2019 (COVID-19), with a cumulative confirmed case of 765 million, including a 1% death rate . In addiThe American College of Rheumatology/European Alliance of Associations for Rheumatology (ACR/EULAR) 2022 classified AAV into granulomatous polyangiitis (GPA), eosinophilic granulomatous polyangiitis (EGPA), microscopic polyangiitis (MPA), and renal-limited vasculitis (RLV) ,13. HoweTwo independent authors conducted systematic literature by searching relevant full-text articles in the PubMed, SCOPUS, and Cochrane library databases up to 1 January 2023. Keywords were  AND . The eligible articles included the full-text report in the English language of biopsy-proven ANCA-GN with pauci-immune deposit by immunofluorescence (IF) study (defined as negative or weak [≤1+] staining of Ig and complement) or by electron microscopy (EM) (defined as no or faint electron-dense deposit [EDDs]) after COVID-19 vaccination. Patients’ age, vaccine types, and the disease onset from the vaccination were not restricted. The exclusion criteria were AAV without GN, ANCA-GN with a non-available IF/EM report, and AAV presenting or having disease flare before the onset of vaccination. We also included our 3 GN-ANCA cases diagnosed and treated at King Chulalongkorn Memorial Hospital, Bangkok, Thailand, from January 2022 to December 2022. Data extraction was anonymized under the General Data Protection Regulation , and infData included patient demographics, vaccine types, vaccination numbers, the interval between the last vaccination and onset of AAV, the clinical manifestation of ANCA-GN and AAV, serum creatinine (Scr) at baseline and peak, serologies , kidney pathology finding, treatment, and outcomes after treatment.Severe acute kidney injury (AKI) was defined as dialysis requirement or peak Scr > 5.7 mg/dL. Outcomes were classified into remission with relapse, remission without relapse, and non-remission . Remission of ANCA-GN/AAV was defined according to the 2012 KDIGO Guidelines for Management of Glomerular Diseases  as the at-test. The continuous data were illustrated by using mean ± standard deviation (SD) for normal distribution data and median (interquartile range [IQR]) for non-normal distribution data. The categorical data were described as a ratio. Finally, the normally distributed variables were compared by A total of 2956 relevant records were initially obtained from the database search. After removing 312 duplicated records, 2644 record titles and abstracts of articles were first screened according to the inclusion criteria. 68 articles were eligible for the second screening; 43 articles (154 cases) were excluded ; 26 cases from 25 articles, including our 3 cases were recruited for the systematic review . The detThe median (IQR) age was 70 (22) years, with female predominance (59%). Most cases presented ANCA-GN after receiving the second vaccination (59%) with a median interval gap of 14 (11–27) days. The mRNA COVID-19 vaccine was the most common (55%), followed by the viral vector vaccine (31%) and inactivated COVID-19 vaccine (14%). The top-three most common presentations were AKI, abnormal urine sediments, and constitutional symptoms. Extra-kidney manifestations included pulmonary involvement  , neuromuscular involvement  , otologic/optic involvement , and Wallerian degeneration . Severe AKI was observed in 34% (10/29). MPO-ANCA/pANCA was positive in 79% (23/29). A discrepancy of ANCA serologies was observed in one case . A dual-positive ANCA (pANCA and cANCA) was detected in 3% (1/29). Autoantibodies were positive in 52% (13/25 cases), including ANA , Coombs antibody , cryoglobulin , and RF . The 2021 KDIGO Guidelines for Management of Glomerular Diseases  were adoWe described three cases of ANCA-GN following viral vectors (case #1) and mRNA (case #2 and #3) COVID-19 vaccines. Only one patient (case #2) was classified as microscopic polyangiitis (MPA) by ACR/EULAR 2022 . Case #19/L with a neutrophil predominance (83%), Scr 3.5 mg/dL, and albumin 2.5 g/dL. Urinalysis demonstrated similar profiles with dysmorphic erythrocytes. The urine protein-creatinine ratio (UPCR) was 7.5 g/g creatinine. pANCA was positive with negative MPO-ANCA and PR3-ANCA. Serologies were positive for ANA , RF 30 (0–20) IU/mL, and Coombs antibody (1+), while others were negative . Complement levels were within normal values. The kidney biopsy disclosed 24 glomeruli with 7 global sclerosis (GS) and 7 cellular crescents. Fibrinoid necrosis of arteries and severe tubulointerstitial inflammation were observed. Interstitial fibrosis and tubular atrophy involved 30% of cortical tissue. IF revealed IgG (1+), C3 (1+), Kappa (1+), and Lambda (1+). Focal trivial subepithelial and intramembranous EDDs with diffuse thickening of GBM were observed on EM. Hence, ANCA-GN without extra-kidney organ involvement was diagnosed. Three-day of 1 gm/day pulse methylprednisolone and 500 mg pulse cyclophosphamide was started. Unfortunately, her kidney function remained deteriorated, requiring maintenance hemodialysis.A 76-year-old female with type 2 diabetes mellitus, hypertension, and dyslipidemia presented with leg edema for 11 days prior to admission. She received the second COVID-19 vaccine (AZD1222) 3 months ago. A few weeks later, she developed chronic fever, weight loss, and productive cough. Several blood cultures were performed, yielding negative results. Non-contrast chest computed tomography (CT) revealed unusual interstitial pneumonia (UIP). Sputum cultures depicted normal flora. She denied a history of SARS-CoV-2 infection and had negative PCR for COVID-19. Her Scr was 1.5 mg/dL over the past year. Three-week before admission, her Scr increased to 1.7 mg/dL with 2+ proteinuria and erythrocyte of 10–20/HPF. A week later, she developed leg edema and foamy urine. Physical examination (PE) revealed temperature 38.9 °C, respiratory rate (RR) 20/min, blood pressure (BP) 165/91 mmHg, and bilateral 3+ leg pitting edema. Blood chemistries revealed hemoglobin 7.6 g/L, leukocyte count of 11.9 × 10Case #29/L with neutrophils predominance (83%), and platelet counts 386 × 109/L. Urinalysis depicted trace protein, erythrocytes 3–5 cells/HPF, and UPCR 0.5 g/g creatinine. pANCA and MPO-ANCA were positive, with negative PR3-ANCA and cANCA. Serologies were positive, including ANA , cryoglobulin, and 1+ direct antiglobulin test, while the others were negative . Complement levels were within normal values. Serum protein electrophoresis and immunofixation demonstrated polyclonal gammopathy. No structural kidney abnormalities were detected by ultrasonography. The kidney biopsy disclosed 13 glomeruli with 2 GS and 10 cellular crescents. Fibrinoid necrosis was found on the vasculitic medium-sized artery. Mild tubulointerstitial fibrosis was observed. IF revealed non-specific staining for all immunoreactants. Hence, AAV with ANCA-GN was diagnosed. She received plasma exchange ×7 times, pulse methylprednisolone, and oral cyclophosphamide. One year later, she remained well without dialysis requirements and active AAV manifestation. Her serum creatinine 2.3 mg/dL and negative erythrocyturia.A 69-year-old female presented with AKI while investigating anorexia, weight loss, and fatigue. She had hypertension for 2 decades and received 2 doses of AZD1222 and a COVID-19 vaccine booster with BNT162b2 (3 months ago). One month later, she developed anorexia, fatigue, a weight loss of 8 kg within 2 months, and established mixed conductive and sensorineural hearing loss. Chest X-ray and contrast computerized tomography (CT) revealed a subcentimeter pulmonary nodule. Two weeks after CT, her Scr increased from 0.7 to 2.4 mg/dL, and contrast-associated AKI was suspected. However, despite receiving an intravenous fluid infusion, she had a steadily increasing Scr. Her urinalysis revealed 1+ protein, leukocyte 50–100/HPF, and erythrocyte 3–5/HPF. Ciprofloxacin was prescribed on suspicion of having a urinary tract infection. PE revealed temperature 38.0 °C, BP 120/60 mmHg, marked pale conjunctivae, and no leg edema. Blood chemistries were notable for Scr 7.1 mg/dL, albumin 2.5 g/dL, hemoglobin 6.6 g/dL, leukocyte counts 12.1 × 10Case #39/L with neutrophils predominance (80%), and platelet counts 209 × 109/L. Urinalysis depicted 2+ protein, leukocytes 30–50/HPF, erythrocytes 1–2/HPF, and UPCR 1.2 g/g creatinine. cANCA and MPO-ANCA were positive, with negative PR3-ANCA. Positive direct antiglobulin test at 1+ was detected, while others were negative . Complement levels were normal. Serum protein electrophoresis and immunofixation demonstrated polyclonal gammopathy. No structural kidney abnormalities were detected by ultrasonography. The kidney biopsy disclosed 20 glomeruli with 2 GS and 7 cellular crescents. Fibrinoid necrosis was noted in 6 glomeruli. Diffuse tubulointerstitial inflammation with abundant eosinophils and trivial tubulointerstitial fibrosis (<5%) were observed. IF revealed non-specific staining for all immunoreactants. ANCA-GN with eosinophilic interstitial nephritis was diagnosed. Due to the alteration of consciousness, magnetic resonance imaging (MRI) of the brain and orbits were performed, demonstrating Wallerian degeneration of the right corticospinal tracts and pons and bilateral optic neuritis. Intravenous pulse methylprednisolone and intravenous immunoglobulin were administered as per AAV. Temporary hemodialysis was initiated for 2 weeks. Subsequently, she remained well without AAV clinical flare with low-dose prednisolone and immunosuppressive agents; her Scr was 1.6 mg/dL with negative proteinuria and bland urine sediment.An 84-year-old female with underlying diseases of hypertension and dyslipidemia presented with a fever for one week prior to admission. She received complete primary series of AZD1222 and a booster dose of the COVID-19 vaccine with mRNA-1273 four months ago. One month later, she developed bilaterally severe sensorineural hearing loss and tinnitus. A few weeks before admission, she developed drowsiness, progressive dysphagia, decreased urine volume, and a weight loss of 8 kg. PE revealed BP 154/66 mmHg, temperature 38.5 °C, mildly pale conjunctivae, right eye episcleritis with afferent pupillary defect (RAPD), and 1+ leg edema. Blood chemistries were notable for Scr 5.1 mg/dL (baseline 0.9 mg/dL), albumin 2.7 g/dL, hemoglobin 9.6 g/dL, leukocyte counts 26.0 × 10ANCA-GN secondary to COVID-19 vaccination was more prevalent in the patients with mRNA vaccinations , older age, and female gender. The top three most common manifestations were AKI, abnormal urine sediment, and constitutional symptoms. MPO-ANCA was distinctly dominant compared to the other ANCAs. Concomitant autoantibodies were documented in 52%, particularly Coombs antibody, ANA, and cryoglobulin. Although one-third (34%) presented with severe ANCA-GN, most cases had good outcomes, with a remission rate of 89%. No death occurred. The higher immunogenicity and more common usage of the mRNA COVID-19 vaccine might explain its higher prevalence on ANCA-GN ,19. The Most AAVs have unknown etiology, known as primary AAVs; however, AAVs can be secondary to drugs  ,25,26,27in-vitro experiment and found that the influenza vaccine (RNA vaccine) was able to induce PR3-ANCA production from peripheral blood mononuclear cells (PBMCs), while the decoy molecule, inactivated split virion influenza vaccine 2007, could not [AAV/ANCA-GN induced by vaccination has been well documented in post-influenza vaccination ,32. The ould not . In addiould not ,34. Unfoould not . COVID-19 vaccine induces ANCA-GN and AAV, possibly through molecular mimicry, stimulating productions of MPO-ANCA and PR3-ANCA via the adaptive immune system, likewise inducing antiviral neutralizing antibodies ,37,38. TTreatment of ANCA-GN following the COVID-19 vaccines in this systematic review mostly complied with the 2021 KDIGO Guidelines , wherebyANCA-associated glomerulonephritis following the COVID-19 vaccine is rare, according to billions of vaccine doses that have been administered worldwide. The benefits of COVID-19 vaccination outweigh the risks of possible induced autoimmune diseases and ACNA-GN. Avoid unnecessary booster doses of the COVID-19 vaccine in high-risk patients  should warrant.


South African Journal of Physiotherapy 78(1), a1611. https://doi.org/10.4102/sajp.v78i1.1611, there was an error regarding the affiliations for the authors. Instead of:In the published article, Anwar, S., Arsalan, S.A., Zafar, H., Ahmed, A., Gillani, S.A. & Hanif, A., 2022, ‘Effects of breathing re-education on endurance, strength of deep neck flexors and pulmonary function in patients with chronic neck pain: A randomised controlled trial’, Authors:‘1https://orcid.org/0000-0002-6603-3185Sahreen Anwar1https://orcid.org/0000-0003-3492-7050Syed A. Arsalan1,2https://orcid.org/0000-0001-9318-3691Hamayun Zafar1https://orcid.org/0000-0002-1965-6224Ashfaq Ahmed3https://orcid.org/0000-0002-2996-0764Syed A. Gillani4https://orcid.org/0000-0002-2670-6402Asif HanifAffiliations:1Department of Physical Therapy, Faculty of Rehabilitation Sciences, University of Lahore, Lahore, Pakistan2Department of Physical Therapy, Faculty of Rehabilitation Sciences, King Saud University, Riyadh, Saudi Arabia3Department of Biostatistics, Faculty of Rehabilitation Sciences, University of Lahore, Lahore, Pakistan’It should be:‘Authors:1https://orcid.org/0000-0002-6603-3185Sahreen Anwar1https://orcid.org/0000-0003-3492-7050Syed A. Arsalan1,2https://orcid.org/0000-0001-9318-3691Hamayun Zafar1https://orcid.org/0000-0002-1965-6224Ashfaq Ahmed3https://orcid.org/0000-0002-2996-0764Syed A. Gillani4https://orcid.org/0000-0002-2670-6402Asif HanifAffiliations:1University Institute of Physical Therapy, University of Lahore, Lahore, Pakistan2Faculty of Rehabilitation Sciences, King Saud University, Riyadh, Saudi Arabia3Faculty of Allied Health Sciences, University of Lahore, Lahore, Pakistan4University Institute of Public Health, University of Lahore, Lahore, Pakistan’.In addition, there was an error on page 2. The following paragraph is updated as it was incorrectly formulated:The original incorrect wording:n = 34 in each group, Z1-α/2 = standardised level of significance = 95% = 1.96, Z1-β = Power of test = 80% = 1.28, µ1 = Mean in control group = 4.60, µ2 = Mean in physiotherapy group = 5.40, δ12 = standard deviation in control group = 0.84, δ22 = standard deviation in physiotherapy group = 0.59. The primary outcome used to estimate sample size was vital capacity (VC) (Duymaz 2019). A difference of 0.5 in the standard deviation was regarded as a meaningful change .Here, The revised and updated wording:n = 15 in each group, Z1-α/2 = standardised level of significance = 95% = 1.96, Z1-β = power of test = 80% = 1.28, µ1 = mean in control group = 4.60, µ2 = mean in physiotherapy group = 5.40, δ12 = standard deviation in control group = 0.84, δ22 = standard deviation in physiotherapy group = 0.59. The primary outcome used to estimate sample size was vital capacity (VC) (Duymaz 2019). A difference of 0.5 in the standard deviation was regarded as a meaningful change .Here, The authors apologise for these errors. The corrections do not change the study’s findings of significance or overall interpretation of the study’s results or the scientific conclusions of the article in any way.


Nonspecific presentation and limited diagnostic solutions of acute infections and sepsis in emergency departments (EDs) result in early antimicrobial administration at a cost to antimicrobial stewardship. We validated the host response classifiers, IMX-BVN-3b and SEV-3b, to determine bacterial and viral infection status and illness severity. We also assessed antibiotic over- and underuse.We prospectively enrolled adult ED patients with suspected acute infections or sepsis with ≥1 vital sign abnormal across 7 sites. At enrollment, we surveyed treating physicians on infection probability and antibiotic prescribing. BVN/SEV-3b were calculated using NanoString nCounter® on PAXgene® blood samples. We compared BVN-3b likelihood of bacterial and viral infection to post-hoc clinical adjudication infection status and SEV-3b likelihood of severe outcomes to 7-day need for ICU care and 30-day mortality.Of 568 enrolled patients, 346 had consensus adjudications . The BVN-3b area under the receiver operating curve (AUROC) for bacterial infections was 0.82 (95%CI 0.77-0.87), compared to 0.76 (95%CI 0.71-0.81) for procalcitonin (p = 0.011). BVN-3b AUROC for viral infections was 0.89 (95%CI 0.84-0.95). SEV-3b AUROC was 0.78 (95%CI 0.69-0.86) and 0.88 (95%CI 0.73-1) for 7-day ICU care and 30-day mortality, respectively. Of the 346, 226 patients had a pre-lab result physician questionnaire; of these, 110 patients received antibiotics. Of these, BVN-3b would have corrected 71% of antibiotic prescribing errors (34/48 overprescriptions and 13/18 underprescriptions).BVN-3b and SEV-3b accurately identified bacterial and viral infections and risk status. If implemented in a rapid workflow, these tests may improve patient management and antibiotic prescribing, reducing healthcare costs in the ED.Arianna Beltran, BS, Inflammatix: Internship Uan-I Chen, MS, Inflammatix: Stocks/Bonds Edward A. Michelson, MD, Inflamatix: Research expense reimbursement for sponsored clinical trial Alexandra Weissman, MD, Inflammatix Inc: Advisor/Consultant Evangelos J. Giamarellos-Bourboulis, PhD, D(ABMM), Abbot Product Operations AG: Grant/Research Support|Abbot Product Operations AG: Honoraria|bioMerieux: Grant/Research Support|bioMerieux: Honoraria|Horizon 2020 European Program: Grant/Research Support|Horizon Health European Program: Grant/Research Support|Sobi AB: Advisor/Consultant|Sobi AB: Grant/Research Support|Sobi AB: Honoraria Oliver Liesenfeld, MD, Inflammatix Inc.: Ownership Interest Natalie N. Whitfield, PhD, D(ABMM), GenMark Dx/Roche: Employee|GenMark Dx/Roche: Stocks/Bonds|Inflammatix: Employee|Inflammatix: Stocks/Bonds


The 13-valent pneumococcal conjugate vaccine (PCV13) was introduced for infant use in the United States in 2010, replacing PCV7. A U.S. case-control study (2010–2014) demonstrated vaccine effectiveness (VE) for ≥1 dose of PCV13 at 86%; however, this study lacked statistical power to examine VE by number of doses and against individual serotypes.We used the indirect cohort method to estimate VE of PCV13 against vaccine-type invasive pneumococcal disease among children < 5 years in the U.S. from May 1, 2010 to December 31, 2019. We included IPD cases identified through CDC’s Active Bacterial Core surveillance in 10 U.S. states; during 2010 – 2014, we additionally included cases enrolled in a post-licensure matched case-control study from expanded sites. Cases and controls were defined as individuals with PCV13-type-IPD (VT-IPD) and non-PCV13-type-IPD (NVT), respectively; serotype 6C was categorized as VT. We used logistic regression to estimate the adjusted odds of prior PCV13 receipt, controlling for confounders identified a priori including age category, race/ethnicity, sex, state, year, and any immunocompromising and/or chronic conditions.>3 PCV13 doses; 47 cases (21.1%) and 53 controls (5.7%) had received no PCV doses. Serotypes 19A (N=96), 3 (N=60), and 19F (N=45) caused 90.1% (201/223) of VT-IPD. VE of >1 or ≥3 PCV13 doses against VT-IPD was 81.7%  and 87.8% (75.2–94.0%), respectively. VE of ≥3 PCV13 doses was 87.0% (75.8–93.0%), 54.6% (-8.8–81.0%), and 92.9% (74.4–98.0%) against serotypes 19A, 3, and 19F, respectively. VE was 87.6% (67.9-95.2%) for three primary doses before 12 months of age and 92.4% (78.2–97.2%) for three primary doses and a booster at 12 months of age or older.A total of 1,161 IPD cases were identified; 223 (19.2%) were VT cases and 938 (80.8%) were non-VT controls. Of those, 108 cases  and 600 controls  had received Vaccine effectiveness estimates against PCV13-type IPD among US children under five years of age, 2010 - 2019VE of ≥1 PCV13 dose against IPD was consistent with the previous estimate from the case-control study, and ≥3 doses appear to provide substantial protection. Among the most commonly circulating VT-IPD, PCV13 was protective against serotypes 19A and 19F IPD; protection against serotype 3 IPD did not reach statistical significance.Lee Harrison, MD, GSK: Advisor/Consultant|Merck: Advisor/Consultant|Pfizer: Advisor/Consultant|Sanofi: Advisor/Consultant Tamara Pilishvili, PhD MPH, GSK: Employed by GSK since February 2023. At the time of data collection and analysis for this work was employed by the CDC


T/C authorization in the US for PrEP of COVID-19 in IC individuals was initially based on a randomized trial (PROVENT). However, < 5% of enrollees in PROVENT were IC. We sought to assess real-world effectiveness of T/C PrEP among IC pts.We conducted a retrospective, observational, propensity-score matched cohort study at UPMC from Jan 1, 2022, to Mar 31, 2023. We assessed effectiveness at ≤ 6 months of T/C 600 mg  against chart confirmed COVID-19 hospitalizations and COVID-19–related deaths among IC pts.Table 1). During the study period encompassing circulating variants both susceptible and resistant to T/C, there were 15 vs 18 COVID-19 hospitalizations across 377,832 person-days in T/C vs non-T/C pts (Table 2). Thus, 0.72% vs 0.87% of T/C exposed vs unexposed pts were hospitalized for COVID-19  . There was no difference in effectiveness by T/C dosing pattern (Table 2). Effectiveness was numerically higher in the BA.5 period, although there were few events in other variant eras (Table 2). There was a non-significant 50% lower risk of hospitalization in T/C patients with solid tumors or autoimmune conditions, and no significant difference by exposure status in patients with organ transplant or hematologic cancer (Table 2). There were 5 COVID-19 ICU admissions per exposure group, and 2 inpatient COVID-19–related deaths (1 per group) (Table 3). Limitations included overall low event rate limiting power and an open healthcare system leading to possibility of missing hospitalizations due to pts seeking care elsewhere.Before matching, there were 2931 T/C vs 157,225 non-T/C pts. After matching, 2301 matched pairs remained . Most pts had moderate/severe IC conditions : Advisor/Consultant|LabSimply: Advisor/Consultant|Merck: Advisor/Consultant|Shionogi: Advisor/Consultant|Shionogi: Honoraria John W. Mellors, MD, AstraZeneca: Grant/Research Support|Gilead Sciences: Grant/Research Support


Their structures and stereochemistry were determined from analysis of NMR spectroscopic and circular dichroism (CD) data. Compound 2 represents the first report of naturally occurring arylnaphthalide lignan triglycoside. The cytotoxic activities of all isolated compounds were evaluated against A-549 and SMMC-7721 cell lines. Compounds 7–10 and 14–16 were more toxic than cisplatin in two tumor cell lines. This investigation clarifies the potential effective substance basis of D. versipellis in tumor treatment.One new dibenzyltyrolactone lignan dysoslignan A ( Arylnaphthalide lignans have received much attention due to their potent antiviral, antineoplastic, anti-inflammatory, and immunosuppressive properties . The repDysosma versipellis (Hance) M. Cheng ex Ying, belonging to the family of Berberidaceae, is widely distributed in the central/south regions of China + ; NMR data (DMSO-d6), see Dysoslignan A (2): white, amorphous powder; + ; NMR data (DMSO-d6), see Dysoslignan B (3): white, amorphous powder; UV (MeOH) λmax (log ε) 204 (4.49), 225 (4.19), 264 (4.35), 326 (3.76), 363 (3.60) nm; IR (iTR)νmax 3367, 2989, 2946, 2833, 1741, 1608, 1520, 1467, 1420, 1348, 1274, 1213, 1186, 1115, 1090, 1027 cm−1; HR-ESI-MS (positive): m/z 385.0920 [M + H]+ , m/z 407.0737 [M + Na]+ ; NMR data (DMSO-d6), see Dysoslignan C (4): white, amorphous powder; UV (MeOH) λmax (log ε) 202 (4.48), 225 (4.29), 263 (4.38), 312 (3.82), 355 (3.60) nm; IR (iTR)νmax 3410, 2939, 2839, 1745, 1605, 1535, 1465, 1352, 1244, 1131, 1094, 1030 cm−1; HR-ESI-MS (positive): m/z 383.0768 [M + H]+ ; NMR data (DMSO-d6), see Dysoslignan D (1 (1.0 mg) and 2 (1.0 mg) were dissolved in 1.0 mL of 2M HCl, and then hydrolyzed at 90 °C for 3 h. The HCl in the reaction mixture was removed under reduced pressure. The remaining reaction mixture was extracted with CH2Cl2. The water layers were directly analyzed by HPLC . The peaks at 13.15 and 14.27 min were coincided with D-glucose and D-galatose.The absolute configurations of the galatose and glucose moieties were determined by the previously reported method . Compoun1–18 were evaluated against human lung cancer A-549, hepatocellular carcinoma SMMC-7721 cell lines. The cells were cultured in RPMI-1640 medium, supplemented with 10% fetal bovine serum (FBS) at 37 ℃ under 5% CO2 in a humidified atmosphere. Cell viability was assessed by conducting colorimetric measurements of the amount of insoluble formazan formed in living cells based on the reduction of 3--5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS). To be brief, 100 µL of cells were seeded into each well in a 96-well cell culture plate in advance. After 24 h, various concentrations of all test compounds were added. After the incubation for 48 h, MTS (20 μL) was added to each well, and the incubation continued for 4 h at 37 ℃. The optical density at 492 nm was determined using a 96-well microtiter plate reader. The IC50 values were calculated by the Reed–Muench method. Statistical analysis were performed by SPSS 20.0 software . All experiments were performed in triplicate.By the previously reported MTS method , the cytD. versipellis resulted in the isolation of one new dibenzyltyrolactone lignan dysoslignan A (1), three new arylnaphthalide lignans dysoslignan B–C (2–4), along with fourteen known metabolites (5–18). Compound 2 is the first reported example of naturally occurring arylnaphthalide lignan triglycoside. All isolated compounds were tested for their in vitro cytotoxic activity against A-549 and SMMC-7721 cell lines using MTS assay. Among them, compounds 7, 14, and 15 were cytotoxic, with IC50 values of less than 1μM. Our research further demonstrated that the arylnaphthalide lignans are mainly responsible for the potent anticancer effect of D. versipellis.Further phytochemical studies on


Staphylococcus/Mammaliicoccus spp. isolates from contralateral healthy skin and DFU from 44 hospitalized patients.Diabetic foot infections (DFIs) are the most common complications of diabetic foot ulcers (DFUs), and a significant cause of lower extremity amputation. In this study we used whole genome sequencing to characterize the clonal composition, virulence and resistance genetic determinants of 58 S. aureus (n = 32) and S. epidermidis (n = 10) isolates were recovered from both DFUs and healthy skin, whereas, S. haemolyticus (n = 8), M. sciuri (n = 1), S. hominis (n = 1) and S. simulans (n = 3) were recovered exclusively from healthy skin. In contrast, S. caprae (n = 2) and S. saprophyticus (n = 1) were recovered only from DFUs. Among S. aureus isolates, MRSA were present with high prevalence , 18 of which (66.7%) were from DFUs and 9 (33.3%) from healthy skin. In contrast, the coagulase-negative Staphylococcus (CoNS)/Mammaliicoccus isolates (n = 26), in particular S. epidermidis and S. haemolyticus were more prevalent in healthy skin,  and , respectively. MLST, spa and SCCmec typing classified the 32 S. aureus isolates into 6 STs, ST672, ST80, ST241, ST1, ST97, ST291 and 4 unknown STs (STNF); 8 spa types, t044, t037, t3841, t1247, t127, t639, t937 and t9432 and 2 SCCmec types, type IV and type III(A). Among CoNS, the S. epidermidis isolates belonged to ST54, ST35 and ST640. S. haemolyticus belonged to ST3, ST25, ST29, ST1 and ST56. The sole M. sciuri isolate was found to carry an SCCmec type III(A). A wide range of virulence genes and antimicrobial resistance genes were found among our isolates, with varying distribution between species or STs. The pan-genome analysis revealed a highly clonal population of Staphylococcus isolates, particularly among S. aureus isolates. Interestingly, the majority of S. aureus isolates including MRSA, recovered from the healthy skin and DFUs of the same patient belonged to the same clone and exhibited similar virulence/resistance genotype.Staphylococcus/Mammaliicoccus spp. in DFIs, which could serve as a basis for further studies on these as well as other groups of pathogens associated with DFIs.Our study provides clinically relevant information on the population profile, virulence and antibiotic resistance of The online version contains supplementary material available at 10.1186/s12866-023-03087-2. Diabetes is a fast-growing global problem with huge social, health, and economic consequences . The prePeople with diabetes are at increased risk of long-term complications such as coronary heart disease, cerebrovascular and peripheral vascular diseases and diabetic foot ulcers (DFUs) . It was Microbial infections of the DFUs, termed diabetic foot infections (DFIs) are key contributors to the amputation risk . Limb loStaphylococcus are common colonizers of skin and mucous membranes of humans and animals, but also opportunistic pathogens capable of causing a wide range of infections. The staphylococci can be differentiated into Coagulase-Positive (CoPS) and Coagulase-Negative (CoNS), based on their ability to produce coagulase. Recently, five species among the CoNS , and belonging to S. sciuri group, were reclassified into the novel genus Mammaliicoccus, with Mammaliicoccus (M.) sciuri as the type species , Staphylococcus/Mammaliicoccus species were identified among the 58 isolates, including S. aureus , S. epidermidis , S. haemolyticus , S. simulans , S. caprae , S. hominis, S. saprophyticus and M. sciuri . The characteristics of the 58 isolates included in this study are shown in Table Eight S. aureus isolates, 23/32 (71.9%) were recovered from DFUs and 9/32 (28.1%) from healthy skin. The 10 S. epidermidis isolates were recovered from 8 patients, including 6/10 (60%) from healthy skin and 4/10 (40%) from DFUs. All the S. haemolyticus (n = 8), M. sciuri (n = 1), S. hominis (n = 1) and S. simulans (n = 3) isolates were recovered exclusively from healthy skin, but S. caprae (n = 2) and S. saprophyticus (n = 1) were recovered only from DFUs.Among the 32 S. aureus isolates, 27/32 (84.4%) carried the mecA gene, and were therefore MRSA; 18/27 of which (66.7%) were from DFUs and 9/27 (33.3%) from healthy skin.Among the 32 S. aureus including MRSA isolates and CoNS were significantly associated with the source of isolation .Statistically, S. aureus were characterized based on the combination of MLST, SCCmec and spa typing. In silico determination of MLST revealed that S. aureus isolates belonged to 6 known STs including ST80 , ST241 , ST672 , ST1 , ST97 and ST291 , and 4 STNF .Clones of S. aureus isolates. The dominant one was t044 , followed by t037 , t3841 , t1247 and t127 , t639, t9432 and t937 .Eight spa types were identified among mec types were identified among MRSA isolates, type IV  and type III(A) . The type IV isolates were assigned to subtype IVc(2B)  and IVd(2B) .Two SCCMammaliicoccus isolates, SCCmec type III(A)  was detected in the sole M. sciuri isolate.Among CoNS/The dominant MRSA clone  was ST80- t044- IVc(2B), followed by ST241-t037- III(3 A)) ; whereas, ST672- t3841- IVd(2B) and STNF- t037- III(3 A) were each represented by 3/27 (11.1%) isolates. In addition, 2 other spa types were detected among ST80- IVc(2B) isolates, t1247  and t639 .2/5 (40%) of MSSA isolates belonged to ST1-t127, while ST97-t9432, ST291-t937 and STNF-t037 were each represented by one isolate .S. epidermidis isolates, 6/10 (60%) belonged to ST54, 2/10 (20%) to ST35 and 1/10 (10%) to ST640. 3/8 (37.5%) S. haemolyticus belonged to ST3 and 2/8 (25%) to ST25, while ST29, ST1 and ST56 were each represented by 1/8 (12.5%) isolate.Among The presence and distribution of the virulence genes are summarized in Tables 2,S. aureus isolates including 42 adhesion genes and a large number of type 8 capsular polysaccharide, immune evasion and exoenzyme genes.A total of 116 virulence genes were detected among hlgA, hlgB, hlgC, hlb, hld and hla/hly), 13 staphylococcal enterotoxins (se) and staphylococcal enterotoxin-like toxins (sel) , with different carriage proportions ranging from 3.1 to 100%. ST1 isolates carried the highest number (n = 7) of se/sel genes, sea, seb, seh, sek, seq, selk and selq. In contrast, none of the se/sel genes were detected among the ST80 isolates.Thirty-five toxin-encoding genes were found among MRSA/MSSA isolates including 6 hemolysins  or a toxic shock syndrome toxin (tst) gene.Remarkably, none of the lukD, lukE) were detected in all S. aureus isolates , while lukF/lukS-PV were detected only in ST80 isolates . In addition, edinB gene encoding epidermal cell differentiation inhibitors was also detected only in ST80 isolates  and in ST291- t937 .The leukocidins genes , which was exclusively present in S. epidermidis isolates .On the other hand, the sole virulence factor found among the 26 CoNS/S. aureus ST or CoNS/Mammaliicoccus species was statistically significant. In contrast, no statistically significant association was found between the presence/absence of virulence genes and PEDIS grades or the source of isolates .The association of virulence genes with S. aureus is shown in Tables mecA and blaZ, were detected among S. aureus isolates at frequencies of 84.4% (27/32) and 50% (16/32), respectively.The distribution of the genetic determinants of antibiotic resistance among the AME), aph(3’)- III/ aph(3’)- IIIa, were the most prevalent among S. aureus isolates . All ST80 isolates  were positive for ant(6)-Ia and aph(3’)-III/ aph(3’)-IIIa genes, and all ST241 isolates  were positive for 3 AME genes, ant(9)-Ia, aac(6’)- aph(2’’) and aph(3’)- III.The genes encoding aminoglycoside-modifying enzymes (S. aureus isolates. The msr(A) and mph(C) genes were detected in all the ST672 isolates , and the erm(A) in all the ST241 isolates .Four genes encoding resistance to macrolide-lincosamide-streptogramin B (MLSB) were detected in S. aureus isolates, tet(M) , tet(K)  and tet(38) . Remarkably, only isolates belonging to t037 and harboring SCCmec-III (ST241 and STNF) carried tet(M).Three genes encoding resistance to tetracycline were detected among fusB and fusC genes coding for fusidic acid resistance were detected in all the ST80-t1247  and ST1-t127 , respectively. The dfrG gene coding for trimethoprim-sulfamethoxazole resistance was detected only in ST241 isolates .The M. sciuri isolates. 18/26 (69.2%) carried both mecA and blaZ genes. Remarkably, the single M. sciuri isolate harbored both mecA and mecA1 . 9/10 (90%) S. epidermidis isolates carried both fusB and fosB genes. In addition, msr(A) and mph(C) were detected in S. epidermidis  and S. haemolyticus  isolates. Moreover, ermC was found in S. haemolyticus  and S. epidermidis  isolates. aac(6’)- aph(2’’)/ aac(6’)-Ie/aph(2’’)-Ia genes were detected in S .haemolyticus  and M. sciuri .As presented in Table aph(3’)-III/ aph(3’)-IIIa  and tet(K)  genes were found exclusively in S. haemolyticus and in S. epidermidis, respectively. Genes conferring resistance to streptogramin (vat(B), vat(C) and vgb(B)), macrolides (vga(B) and vga(A)LC), kanamycin/neomycin (aadD), tetracycline (tet(L)), streptomycin (str) and to quaternary ammonium compounds (qacB) were detected only in S. haemolyticus isolates, at a frequency of one gene per isolate.The S. epidermidis , S. aureus , S. haemolyticus  and M. sciuri  . This is consistent with earlier findings that S. aureus isolates recovered from 4 distinct anatomical sites  of patients with type 2 diabetes were highly related in the same patient  was performed using Roary . Tree cop-value for describing possible associations between species/ST and virulence/resistance genes with PEDIS grades and the source of isolates. p-values < 0.05 were considered statistically significant.Data were analyzed using the Statistical Package for Social Sciences . Contingency tables were constructed and Chi-squared tests were used to calculate Below is the link to the electronic supplementary material.Supplementary Material 1Supplementary Material 2Supplementary Material 3


Correction to: Psychometrika 10.1007/s11336-022-09879-1Second line in Eq. (15): “below Eq. (23): “Second line in Eq. (27): “The original version of the article contains the below listed errors. The following transcription errors have been corrected:


The HVTN 704/HPTN 085 clinical trial (AMP study) assessed the efficacy of VRC01 broadly neutralizing antibody infusions for HIV prevention. It offered access to oral pre-exposure prophylaxis (PrEP) at no-cost to all participants as standard of HIV prevention care. Understanding PrEP initiation among trial participants and describing its impact on study endpoints can guide future HIV prevention trial design.We characterized sociodemographic and geographic features of PrEP initiation (by self-report) using descriptive statistics. We used Cox models to identify factors associated with PrEP initiation and the association between PrEP initiation (as a time-dependent exposure) and HIV incidence.Of 2221 participants off PrEP at enrollment, 31.8% initiated PrEP during the trial. Median time to PrEP initiation was 60 days (range 1-673 days). Brazil had the highest proportion of PrEP initiation among their enrolled participants (83.2%) and Peru had the lowest (5.3%). In US sites, 54.2% initiated PrEP. Southern US sites had the highest proportion of PrEP initiation (68.8%) and the Midwest had the lowest (28.4%).In multivariate analysis, prior PrEP use [HR (95% CI) = 2.2 (1.3-4.0)] was associated with PrEP initiation. Participants from Switzerland [0.5 (0.3-1.0)] and Peru [0.08 (0.06-0.1)] had lower likelihood of PrEP initiation compared to the US, while participants from Brazil had higher likelihood [2.6 (2.0-3.3)]. Age, race, gender, and sexual orientation were not associated with PrEP initiation. HIV incidence rate was 3.6/100 person-years (PY) before PrEP initiation, compared to 1.2/100 PY after PrEP initiation. PrEP initiators had 58% less risk of acquiring HIV after adjusting for behavioral risk score and treatment arm. In the US, higher local PrEP-to-need ratio was also associated with lower PrEP initiation [0.9 per 5 units (0.8-1.0)].Providing PrEP access to AMP participants reduced HIV acquisition among PrEP initiators. Study endpoints were still met due to heterogenous regional PrEP uptake and lower PrEP effectiveness than expected. These data suggest that offering oral PrEP to HIV prevention trial participants as standard of care is a feasible option to consider for future efficacy HIV prevention trial design.Valeria D. Cantos, MD, Gilead Sciences: Grant/Research Support Srilatha Edupuganti, MD MPH FIDSA, Sanofi: Grant/Research Support Colleen F. Kelley, MD, MPH, Gilead Sciences: Grant/Research Support|Humanigen: Grant/Research Support|Moderna, Inc.: Grant/Research Support|Novavax, Inc: Grant/Research Support|Viiv Healthcare: Grant/Research Support


Phage-antibiotic combinations (PAC) have been proposed for high inoculum daptomycin-nonsusceptible (DNS) MRSA infections refractory to conventional therapy. We studied PAC with synergistic activity against two DNS MRSA clinical bloods isolates .9 CFU/mL) using: i) modified checkerboard (CB) minimum inhibitory concentration (MIC); and ii) 24h time-kill assays (TKA). Synergistic activity in CB assays was defined as either: a fractional inhibitory concentration (FIC) index ≤ 0.5 in modified CB assays; or a ≥ 2 log10 CFU/mL reduction by PACs vs the most active single-agent regimen. Significant differences between regimens were assessed by ANOVA with Tukey’s post hoc modification (P < 0.05).PAC containing DAP and/or ceftaroline (CPT) (each at ½ MIC) plus a 2-phage cocktail  were tested at a high MRSA inoculum  plus either DAP or DAP + CPT demonstrated robust synergistic activity  vs. the next most effective regimen of CPT + Intesti13+Sb1 (P < 0.05 each). Against C37, Intesti13 + Sb1 (MOI of 1 and 0.1) with CPT or DAP + CPT were equally potent and effective regimens (-Δ7.14 log10 CFU/mL each), but neither were significantly better than the synergistic regimen of DAP + Intesti13 + Sb1 (-Δ6.65 log10 CFU/mL).By CB assay, synergistic activity was demonstrated with Intesti13 + Sb1 (MOI of 10 to 0.01) plus either DAP or DAP + CPT (FIC ≤ 0.5 for each combination). In 24h TKA vs These studies demonstrated positive phage-antibiotic synergy (PAS), with phage cocktail Intesti13 + Sb1 and either DAP or DAP + CPT. Addition of these phages to DAP or DAP + CPT combinations caused bactericidal and synergistic killing vs. CPT + phages. *, P<0.05. Abbreviations: CFU: colony forming units, GC: growth control, DAP: daptomycin, CPT: ceftaroline, MOI: multiplicity of infection, PAC: phage-antibiotic combination.These studies demonstrate positive phage-antibiotic synergy (PAS) with phage cocktail Intesti13 + Sb1 added to DAP, CPT, and DAP + CPT at an MOI of 1 and 0.1. Abbreviations: CFU: colony forming units, GC: growth control, DAP: daptomycin, CPT: ceftaroline, MOI: multiplicity of infection, PAC: phage-antibiotic combination.in vivo investigations of these candidate PACs, for treatment of high inoculum DNS MRSA infections  is warranted.The two-phage cocktail used (Intesti13 + Sb1) demonstrated impressive synergistic activity against two DNS MRSA isolates in combination with DAP or DAP + CPT. Further experimental Arnold S. Bayer, MD, Akagera Medicines: Grant/Research Support|ContraFect Corporation: Grant/Research Support Michael J. Rybak, PharmD, PhD, MPH, Abbvie, Merck, Paratek, Shionogi, Entasis, La Jolla, T2 Biosystems: Advisor/Consultant


Triticale  is of great interest as an insurance crop that can ensure the stability of the gross harvest of feed and food grains at a lower cost. In Western Siberia, only winter triticale varieties are cultivated, however, spring triticales are important for cultivation in regions not suitable for winter crops. To create spring varieties with high yields and good grain quality, it is necessary to study and enrich the gene pool, identify donors of economically valuable traits. One of the possible ways to solve this problem can be through the production of secondary hexaploid triticales with the involvement of the tetraploid wild-growing species of emmer wheat Triticum dicoccum (Schrank) Schuebl. The aim of this work was to create and study hybrids of emmer T. dicoccum (Schrank) Schuebl. with hexaploid triticale using genomic in situ hybridization for staining of meiotic chromosomes and analysis of plant productivity elements in F4–F8. DT4, DT5, DT6 plants and the prebreeding F6 forms obtained from them – DT 4/168, DT 5/176 and DT 6/186 – were selected according to the characteristics of the productivity and the nature of the grain in the F4 hybrid population. The offspring of hybrids DT4 and DT5 and prebreeding forms DT 4/168 and DT 5/176 had an increased grain nature (over 750 g/l), but low productivity. The hybrid DT6 and the breeding form DT 6/186 obtained from it had high grain productivity , but, like the paternal form of triticale UK 30/33, had a reduced nature of the grain. In F8 DT 6/186 plants, 7 homologous pairs of rye chromosomes and from 27 to 30 wheat chromosomes were found in meiosis, which indicates the presence of a complete rye genome and two wheat ААВВ genomes. Rye chromosomes showed stable formation of bivalents in contrast to wheat chromosomes, which caused the presence of aneuploids in plant populations. Thus, hexaploid forms DT 4/168 and DT 5/176 with well-made smooth grain and high grain size were obtained, which can be used as a source of this trait for selection of food-grade triticale. DT 6/186 is a promising form for further breeding in order to obtain high-yielding forms of triticale. Triticale  is a wheat and rye hybridof a relatively short evolutionary history as an allopolyploidspecies. The first naturally created fertile wheat×rye hybridswere discovered in the late 1920s at the South-Eastern agriculturalexperimental station in Saratov . Theplants had intermediate traits and were described as a new botanicalspecies Triticum Secalotriticum saratoviense Meisterby G.K. Meister . Meister immediately predictedthe practical value of these intergeneric crossings. Thefirst man-made wheat×rye hybrids were obtained in 1888 by the German plant breeder W. Rimpau, who described 12 plants descending froma wheat×rye hybrid generally recognized as the first triticales. The cytological analysis of thefirst triticales developed in Russia and Germany showed thesomatic chromosome number of 56 (8х) , which demonstrated the combinationof four genomes as follows: AABBDDRR, AABBDD,soft wheat, and RR rye.Octoploid triticales were of great interest for plant breedersdue to high seed set per spike, increased plant pathogen resistance,and resistance to environmental stresses. However, theprimary triticale lines suffered from meiotic errors  and high frequency of aneuploidy in earlier generations, which caused reduced fertility. Grain shriveling andlatenessof maturity in octoploid forms were also consideredas limitations for their introduction as cultivars. As a result,there have been global efforts to develop the technology forobtaining more promising lines with various valuable breedingtraits, which eventually produced hexaploid triticales.Primary hexaploid triticales of the AABBRR genome typewere obtained as crossings of tetraploid wheats  and S. cereale rye . However, undesirabletraits were not completely eliminated. Agronomicallyvaluable traits were improved by enriching the triticalegene pool with crossing experiments involving octoploidlines and commercial soft wheat varieties, as well as hexaploidtriticales with full rye genome and two wheat genomesidentified in the progeny of octoploid triticales as a resultof D-genome chromosome elimination . The crossing of two primaryoctoploid triticales produced hexaploid offspring, and breed-ingprograms were focused on hybridization of these octoploidswith the hexaploids identified in the progeny .Hexaploid triticales were more stable in terms of productivity. As a result, various recom-binantforms of secondary triticales were derived and karyotyped,and some of them having combined wheat-rye genomesshowed commercial value .Agronomic traits of triticales were further improved byintergeneric and interspecific crossings. Aegilops crassa , Ae. juvenalis , Ae. squarrosa and Ae. triaristata  , Agropyron intermedium ssp. trichophorum(2n = 42) , Hordeum parodii Covas, H. vulgare L. and T. monococcum L.  plants wereused in hybridization experiments with hexaploid triticales.Intergeneric polyploid triticales were also bred through hybridizationwith intermediate forms sharing at least one setof chromosomes (genome) with the triticale genome. Thesehybridization efforts produced plants with resistance genesagainst diseases. Lines isolated in the progeny of triticale hybrids(AABBRR) and amphidiploids  were resistant toyellow rust . Hybridization of a hexaploidtriticale and an amphiploid intermediate form  produced addition andsubstitution lines with the 3Sv Ae. variabilis chromosome carryingthe powdery mildew resistance gene Pm13 . Addition and substitution lines for chromosome 2Dwith the leaf-rust resistance gene Lr39 and semidwarf geneRht8, as well as chromosome 3D (or 3D/3B) with the Lr32 gene were isolated in the progeny of triticale × amphiploid hybrids .Thanks to breeding achievements, triticale has becomea new economically significant cereal species characterizedby high grain and vegetative mass productivity, that could beused as forage and green feed . In the last three decades, its products have becomeincreasingly significant, which is demonstrated by increasingcrop acreage across the world, from 1,453,269 ha in 1994 to4,157,018 ha in 2016. Triticale grains are used to producebioethanol and food wrap, as well as various food products , andbran is used as the source of prebiotics and antioxidants foryoghurts. Food-grade triticale grains are comparable to wheatin macro- and micronutrient contents . Proteincontent in triticale grains is 1–1.5 % higher than in wheat and3–4 % higher than in rye, gluten content matches that in wheator is 2–4 % higher . However, triticaleunderperforms in test weight. This parameter is closely relatedto the plumpness and hardness of grains, as well as to theirsize and shape. Average test weight for wheat is 700–810 g/l.At test weights below 740 g/l, flour yield tends to decreaserapidly as test weight goes down. Most spring triticales haveshriveling grains and low flour yields, which limits their usein bread making .Development of domestic high-productivity triticale varietieswith high grain quality requires further study and enrichmentof the gene pool, as well as identification of donors foreconomically valuable traits. One way to solve this problemis to obtain secondary hexaploid triticales using emmer wheatT. dicoccum (Schrank) Schuebl., a tetraploid wild-growingwheat with long, large, and plump grains.The goal of the present study was to breed new forms ofhexaploid triticales (ААВВRR genome type) with improvedtest weights by crossing emmer  with triticale and study their productivity and meioticstability with chromosome staining using genomic in situhybridization.Plant material. New forms of hexaploid triticale were obtainedby hybridization of emmer (T. dicoccum (Schrank)Schuebl.) and triticale . IntergenericF1 hybrid of emmer lines (L133 × PKK) × k-25516 (AABBgenome) was used as maternal plants. Awned semi-hullessemmer (L133 × PKK) created by VIR researchers is characterizedby brittle spikes and low productivity, and awnlessemmer k-25516 was obtained at Siberian Research Instituteof Plant Production and Breeding (SRI PPB), ICG SBRAS from the population of awned emmer wheat from theVIR collection. The paternal plants were represented by a selectionform of hexaploid triticale UK 30/33 selected fromthe population of cytogenetically unstable octoploid triticaleUK30 (AABBDDRR genome) developed at the SRI PPB bycrossing the Ulyanovka soft wheat variety with the Korotkostebelnaya69 short stem rye with subsequent doubling ofchromosome number induced by colchicine water solution.The progeny of three F4 hybrids DT4, DT5, and DT6, andthree selection forms DT 4/168, DT 5/176, and DT 6/186 isolatedin F5 from hybrid populations DT4, DT5, and DT6 respectivelywas selected for the study. In 2020, their F6 progenywas seeded for research purposes along with F4 hybrids atthe nursery for distant wheat hybrids in the field of SRI PPB.Fluorescence in situ hybridization of meiotic chromosomes.To evaluate the meiotic stability in the selection forms,two most productive plants DT 6/186/156 and DT 6/186/165were selected in the F7 progeny of plant DT 6/186, and theirF8 seeds were seeded in the hydroponic greenhouse of theICG SB RAS in spring 2021. The chromosomal behavior ofthe progeny was studied using routine acetocarmine stainingprotocol and FISH staining (fluorescence in situ hybridization)following the technique described earlier .The meiocytes were analyzed at the diakinesis stage, metaphaseI (МI), anaphase I (AI), and telophase II (TII). Theprobes used in the analysis were as follows: Aegilops tauschiipAet6-09 specific for centromere repeats in rice, wheat,rye, and barley chromosomes ; pAWRcspecific for centromere repeats in rye chromosomes , and rye genomic DNA. DNA repeat samples ofpAet6- 09 and pAWRc were the courtesy of Dr. A. Lukaszewcki.Centromere-specific probes were labeled with biotin 16-dUTPor digoxigenin 11-dUTP by means of polymerase chain reaction(PCR). The total rye DNA was labeled by Nick translationwith digoxigenin 11-dUTP. The probes were combined indifferent proportions and mixed with blocking wheat DNA.The preparations were mounted in Vectashield antifade solution(Vector Laboratories) slowing down fluorescence fadingand including 1 μg/ml DAPI  for chromatin staining. Allpreparations were analyzed using an Axio Imager M1 microscope, the images were recorded usinga ProgRes MF camera  at the Centerof Microscopic Analysis of Biological Objects, SB RAS andprocessed using Adobe Photoshop CS2 software.Analysis of economically valuable traits. Structuralanalysis of the plants was performed at the facility equippedfor metric measurements, threshing, and seed weighting. Asa result, the following data on productivity elements wereobtained: spike length; spike density; grain weight per spike;thousand kernel weight; grain number per spikelet; testweight measured using a microchondrometer ; and grain productivity per 1 m2. Statisticalprocessing of the results was carried out following thestandard technique . Significance of differencesbetween mean values of two samples was estimatedusing Student’s t-test.The parental-line plants had different spike and grain morphology.The emmer plants (L133 × PKK) had short, awned,brittle spikes and smooth, long grains . The k-25516emmer sample was an awnless plant with thin, long grains . Triticale UK 30/33 had a dense, awned spike and grains similar to those of soft wheat in shape but shriveling.Plants of three emmer×triticale F4 hybrids had differentspike morphology . The DT6 hybridhad a dense, awned spike. The plants of the remaining twohybrids had looser spikes. The spikes were sterile at the endsand therefore often susceptible to ergot.All hybrids had a hair neck similar to the paternal triticaleUK 30/33. This means that these genotypes have a gene responsiblefor manifestation of this trait localized in the longarm of chromosome 5R.Triticale UK 30/33 had a short spike, low test weight, andmedium grain productivity (see Table 1). Selection forms andhybrids, except for DT5, had a longer spike than the paternalform UK 30/33. Hybrid DT5 had low productivity due toflower sterility in the upper part of the spike and low thousandkernel weight. Hybrid DT4 had a high thousand kernelweight of 50 ± 2 g, and the highest test weight of 806 ± 14 g/l. However, its grain productivity, although higher than that ofDT5, was still not too high due to low seed set of spikes andspikelets. Flowers at the top of the spike were often sterile aswell. Hybrid DT6 was characterized by dense spikes and highgrain productivity (785 ± 41 g/m2), as well as seed set of spikesand spikelets. The spikes were fertile along their full length.Selection forms DT 4/168, DT 5/176, and DT 6/186 haddifferent grain weights per spike, grain numbers per spike,test weights, and productivities. DT 4/168 had denser spikesand slightly higher seed sets of spikes and spikelets comparedto the DT4 hybrid it was obtained from. Breeding sampleDT 5/176 was more productive than the initial DT5 hybrid,but it had the lowest productivity among the three breedingsamples studied. Small spike size and sterility of 3–7 spikeletsin the upper part of the spike resulted in low productivityof hybrids DT5 and DT 5/176. The DT 6/186 line had thehighest seed set and productivity among the studied lines.Despite the lowest test weight values similar to triticaleUK 30/33 (706 ± 25 g/l), grain productivity per plot reached822 ± 74 g/ m2. DT 6/186 had a smoother overall morphologythan the initial hybrid DT6, but higher breeding value due toits higher grain productivity.Cytological analysis of the progeny of two plants (DT 6/186/156 and DT 6/186/165) from the highly productive DT 6/186line revealed instability in chromosome number and errors inchromosomal behavior in the first and second meiotic divisions.Chromosome staining using genomic in situ hybridizationin these plants showed 14 rye chromosomes formingbivalents, which implied the presence of seven homologouspairs . However, the bivalent chromosomes demonstratedpremature separation in some meiocytes (desynapsis),with rye chromosomes becoming univalent and distributinganomalously between the poles .Chromosome number in the discovered aneuploid plantsvaried from 2n = 41 to 2n = 44. Among the ten plants from theDT 6/186/156 progeny, only one plant had chromosome number2n = 42 , while there were no plants with euploidchromosome numbers in the progeny of DT 6/186/165.Univalents were discovered in the metaphase I , which were lagging at the equator during chromosomeseparation in anaphase I in 86.75 ± 4.56 and61.32 ± 2.81 % of cells in DT 6/186/156 and DT 6/186/165respectively . The lagging chromosomeswere divided into sister chromatids or broken at the centromere. Instability of chromosome separationduring the division resulted in micronuclei formation atthe tetrad stage . Micronuclei were observed in 60.29 ± 3.14 and 72.16 ±± 2.29 % tetrads in DT 6/186/156 and DT 6/186/165, respectively(see Table 2). Even the euploid plant with 2n = 42(DT 6/186/156) had micronuclei in 51.48 % tetrads, whichprevented us from considering this plant fully cytogeneticallystable.Plants with the minimum number of anomalous meiocytesin anaphase I and telophase II were selected from populationsDT 6/186/156 and DT 6/186/165 based on the results of theanalysis.Triticale  as an agricultural cropcombines wheat’s high yield potential with rye’s resistanceto biotic and abiotic stresses, which increases its adaptability to cultivation conditions in salty or highly acidic soils and inpresence of toxic heavy metals. Thanks to these traits, triticaleis of great interest as an emergency crop ensuring stable grossharvest of forage and food grains at lower costs . Despite today’s applications of triticale grainsbeing mostly restricted to forage in animal husbandry and productionof feed and bioethanol, there is a rising interest in theuse of triticale grains in human food products. Triticale grainshave proven nutritional and dietary value , since they include not only proteins, carbohydrates,and fats, but also vitamins, minerals, and dietaryfibres (14–18 %) . Comparedto that of wheat, the protein from its grains has a richer aminoacid profile, particularly in indispensable amino acids, suchas lysine, threonine, and leucine . Triticale starch amounting to 3/4 of thekernel weight has a significantly lower amylose content comparedto rye and wheat , which ensures its betterdigestion by humans .To increase the share of triticales in production of breadsand pastries, in recent decades breeding efforts have beenaimed at increasing the quality of grains and finished products,which has resulted in development of triticale bread-makingvarieties , so State standards for triticaleflour have been developed (State Standard 34142-2017).The characteristics of winter triticale varieties also includeapplicability for bread making. Triticale breeding efforts inRussia and other countries are primarily aimed at developingwinter varieties . However, grainquality assessment of the samples from the spring triticalecollection showed that spring triticales have good potentialfor creating bread-making varieties . The samples with such improvedparameters as protein content, test weight, falling number,vitreousness, gluten quantity and quality, etc., are used inbreeding for improved grain quality .A trait such as high test weight can be transferred by distanthybridization. This parameter is closely related to geneticallydetermined parameters such as grain plumpness, hardness, andshape. In the present paper, tetraploid emmer wheat T. dicoccum(Schrank) Schuebl. with long, large, and plump grainswas used as the maternal line for hybridization with hexaploidtriticale . As a result, new hexaploidforms of triticale with AABBRR genome types were created,which is confirmed by the analysis of meiotic chromosomalbehavior using genomic in situ hybridization. Seven pairs ofrye chromosomes and 27 to 30 wheat chromosomes were observedin the plants, which implies the presence of a completerye genome and two wheat genomes. Three plants, DT4, DT5,and DT6, were isolated in the F4 population, the progeny ofwhich demonstrated test weight values significantly exceedingthose of the initial triticale line, and test weights in the progenyof DT 4/168, DT 5/176, and DT 6/186 forms from F6 werehigher or on par with those of the initial triticale line. Theseplants had different productivity values, with the highest onesin F4 recorded in the DT6 line (785 ± 41 g/m2). The respectivevalue for DT 6/186, i. e., the progeny of the latter in F6,reached 822 ± 74 g/m2. The DT 4/168 and DT 5/176 lines areof moderate interest for further research into improvement ofbread-making properties of triticale grains.The study of the meiotic behavior of rye and wheat chromosomesin the progeny of F8 plants DT 6/186/156 andDT 6/186/165 showed that they had not yet achieved cytogeneticstability, which was indicated by the discovered chromosomeseparation errors and the presence of aneuploids in thepopulations. Chromosome separation errors mostly occurredin wheat chromosomes due to their monosomy. Cytologicalinstability and aneuploidy in octoploid and hexaploid wheatryeallopolyploids have been an issue from the start , but secondary triticales turned outto be more cytogenetically stable than primary ones . Cytological study of triticales showed that the interactionbetween wheat and rye genomes in the cells of the sameplant resulted in physiological defects in cells persisting fordecades at least. Meiotic and mitotic errors were found bothin the triticale obtained by Rimpau in 1888  and in the triticales obtained later. Despite thecomplete set of chromosomes, meiotic univalents were foundin triticales with different ploidy levels . The studyinto meiotic chromosomal behavior in triticales in the presentand earlier papers demonstrated that while only bivalentswere present at the diakinesis stage, univalents appeared inmetaphase I as a result of to produce lagging chromosomes atthe equator . Presumably, aneu-ploidcells in triticales may emerge as a result of asynchronousseparation of rye and wheat chromosomes and their laggingin anaphase and telophase . Thepresumed cause of meiotic instability in the obtained amphidiploids is an imbalance in the genetic system of meioticpairing and the differences in cell cycles between wheat andrye .It is known for a fact that chromosome separation dependson kinetochore function . It was found thatkinetochore protein CENH3 produced by one of the parentsmaintained the function of other parental kinetochores in stablehybrids, despite the differences between DNA sequences ofcentromere regions in parental lines . It wasalso shown that cytogenetic stability in triticales could also belinked to increased expression of rye-specific CENH3 formsin a hybrid genome .Another possible cause of cytogenetic instability of theobtained triticales could be a nuclear-cytoplasmic incompatibility,because the hybrids are bred using an intergenericF1 hybrid of emmer lines (T. dicoccum (Schrank) Schuebl.)(L133 × PKK) × k-25516 (genome AABB) as the maternal form.The AABB genomes in paternal triticale forms originate fromsoft wheat, while the rye genome is used as the base. Selectionof genotypes of wheat varieties during triticale backcrossingmay lead to recovery of fertility and cytogenetic stability innew forms, as exemplified by alloplasmic soft wheat lines(H. vulgare)-T. aestivum .The result of this work was to create and study hybrids ofemmer T. dicoccum (Schrank) Schuebl. with hexaploid triticale.According to the characteristics of productivity and testweight values of the grain, hexaploid prebreeding forms F6were isolated – DT 4/168, DT 5/176 and DT 6/186. Thus,hexaploid forms DT 4/168 and DT 5/176 with well-filled,smooth grains and high test weights that can be used as thesource of said traits in food-grade triticale breeding havebeen obtained. The DT 6/186 line shows promise in terms ofbreeding for high yields. As a result of the analysis of meioticdivision in DT 6/186 plants, this sample is unstable forwheat chromosomes, therefore, aneuploids will occur in theoffspring. 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