Bio-perfume guns: Antifungal volatile activity of Bacillus sp. LNXM12 against postharvest pathogen Botrytis cinerea in tomato and strawberry.

Gray mold disease, caused by Botrytis cinerea is a major postharvest disease impacting fruits such as strawberries and tomatoes. This study explores the use of volatile organic compounds (VOCs) produced by Bacillus spp. as eco-friendly biocontrol agents against B. cinerea. In vitro experiments demonstrated that VOCs from Bacillus sp. LNXM12, B. thuringiensis GBAC46, and B. zhanghouensis LLTC93-VOCs inhibited fungal growth by 61.2%, 40.5%, and 21.6%, respectively, compared to the control. LNXM12 was selected for further experiments due to its highest control efficacy of 58.3% and 76.6% on tomato and strawberry fruits, respectively. The LNXM12 VOCs were identified through gas chromatography-mass spectrometry (GC-MS) analysis, and 22 VOCs were identified. Synthetic VOCs with the highest probability percentage, namely ethyloctynol, 3-methyl-2-pentanone (3M2P), 1,3-butadiene-N, N-dimethylformamide (DMF), and squalene were used in experiments. The results showed that the synthetic VOCs ethyloctynol and 3M2P were highly effective, with an inhibition rate of 56.8 and 57.1% against fungal mycelium radial growth at 120 µg/mL on agar plates. Trypan blue staining revealed strongly disrupted, deeper blue, and lysed mycelium in VOC-treated B. cinerea. The scanning and transmission electron microscope (SEM and TEM) results showed that fungal mycelium was smaller, irregular, and shrunken after synthetic VOC treatments. Furthermore, the synthetic VOCs Ethyloctynol and 3M2P revealed high control efficacy on tomatoes and strawberries infected by B. cinerea. The control efficacy on leaves was 67.2%, 66.1% and 64.5%, 78.4% respectively. Similarly, the control efficiency on fruits was 45.5%, 67.3% and 46.3% 65.1%. The expression of virulence genes in B. cinerea was analyzed, and the results revealed that selected genes BcSpl1, BcXyn11A, BcPG2, BcNoxB, BcNoxR, and BcPG1 were downregulated after VOCs treatment. The overall result revealed novel mechanisms by which Bacillus sp. volatiles control postharvest gray mold disease.

Therapy-related myeloid neoplasms with normal karyotype show distinct genomic and clinical characteristics compared to their counterparts with abnormal karyotype.

Therapy-related myeloid neoplasms (t-MNs) are a complication of treatment with cytotoxic chemotherapy and/or radiation therapy. The majority of t-MNs show chromosomal abnormalities associated with myelodysplastic syndrome (MDS) or KMT2A rearrangements and are characterized by poor clinical outcomes. A small but substantial subset of patients have normal karyotype (NK) and their clinical characteristics and mutational profiles are not well studied. We retrospectively studied patients diagnosed with t-MN at three institutions and compared the mutational profile and survival data between t-MNs with NK and t-MNs with abnormal karyotype (AK). A total of 204 patients with t-MN were identified including 158 with AK and 46 with NK. NK t-MNs, compared to AK, were enriched for mutations in TET2 (p < 0.0001), NPM1 (p < 0.0001), ASXL1 (p = 0.0003), SRSF2 (p < 0.0001), RUNX1 (p = 0.0336) and STAG2 (p = 0.0099) and showed a significantly lower frequency of TP53 mutations (p < 0.0001). Overall survival (OS) was significantly lower in AK t-MNs as compared to NK cases (p = 0.0094). In our study, NK t-MNs showed a significantly better OS, a higher prevalence of MN-associated mutations and a lower frequency of TP53 mutations compared to their AK counterparts. The distinct clinical and mutational profile of NK t-MNs merits a separate classification.

Therapy-related myeloid neoplasms with single-hit TP53 mutations share the clinical, molecular, and survival characteristics of their multi-hit counterparts.

Recent updates in the classification of myeloid neoplasms (MNs) recognize the poor prognostic impact of TP53 mutations, with particular emphasis on the TP53 allele status. Studies on the effect of TP53 allele status exclusively in therapy-related MNs (t-MNs) are lacking. We compared the clinicopathologic and survival characteristics of t-MNs with single-hit (SH) and multi-hit (MH) TP53 mutations. A total of 71 TP53-mutated t-MNs were included, including 56 (78.9%) MH and 15 (21.1%) SH. Both groups showed comparable genetic profiles with an excess of high-risk karyotypes and a paucity of other co-mutated genes. TP53 was the sole detectable mutation in 73.3% of SH and 75.0% of MH cases. The overall survival (OS) of SH TP53-mutated t-MNs was not significantly different from MH cases (median survival: 233 vs.273 days, p = 0.70). Our findings suggest that t-MNs with SH TP53 mutations share the poor prognostic and biologic profile of their MH counterparts.

Characterization of T-/natural killer cell lymphoproliferative neoplasms associated with systemic, chronic, active Epstein-Barr virus in adults: A report of 5 cases in a Western population.

OBJECTIVES: Because of its low frequency in adult populations and clinical and laboratory overlap with hemophagocytic lymphohistiocytosis and other T-cell lymphomas, T-cell/natural killer (NK) cell systemic, chronic, active Epstein-Barr virus (EBV) (T/NK sCAEBV) infection remains underdiagnosed, preventing critical, prompt therapeutic interventions. METHODS: We report a 5-case series that included 2 adult patients with T/NK sCAEBV and 3 additional adult patients with T/NK lymphomas with concomitant systemic EBV infection to review these entities' overlapping diagnostic and clinical features. RESULTS: Approximately 95% of the world population has been infected with EBV during their lifetime, and infection is usually asymptomatic, with symptomatic cases eventually resolving spontaneously. A small subset of immunocompetent patients develops CAEBV, a life-threatening complication resulting from EBV-infected T-cell or NK cell neoplastic lymphocytes. The sites of end-organ damage in T/NK sCAEBV demonstrate pathologic findings such as reactive lymphoid proliferations, making the diagnosis difficult to establish, with the only curative option being an allogeneic hematopoietic stem cell transplant. CONCLUSIONS: This diagnosis is most prevalent in Asia, with few cases reported in Western countries. Adult age is an independent risk factor for poor outcomes, and most cases are diagnosed in pediatric populations.

Myeloid Sarcoma: Features in Cytological Preparations and the Utility of Rapid On-Site Evaluation (ROSE) in Triage.

INTRODUCTION: Myeloid sarcoma (MS) is a rare extramedullary tumor consisting of blasts of granulocytic, monocytic, erythroid, or megakaryocytic lineage that disrupts the architecture of the involved tissue. MS shows vast clinical, morphologic, immunophenotypic, and genetic heterogeneity posing a diagnostic dilemma, especially in small biopsy specimens such as fine-needle aspiration (FNA) and core biopsy. The objective of this study is to highlight the morphologic features of MS in cytological preparations and investigate the efficacy of pathologist-performed rapid on-site evaluation (ROSE) in assuring accurate triage. METHODS: A retrospective review was performed for cases of MS with concurrent cytology and ROSE results from 2006 to 2017. FNA smears and touch preparations were reviewed, and the results of ROSE, immunohistochemistry (IHC), flow cytometric immunophenotyping (FCI), cytogenetics/FISH, and histology were analyzed. RESULTS: A total of 15 cases were found including 6 (40%) with monocytic morphology comprising promonocytes and monoblasts and 9 (60%) with conventional myeloblastic morphology. The most common genetic subgroup was KMT2A-rearranged MS (33.3%) followed by extramedullary blast crisis of chronic myeloid leukemia (26.6%). ROSE provided sufficient preliminary information and ensured the procurement of adequate tissue for histology, IHCs, FCI, and cytogenetics/FISH, leading to an accurate and complete diagnosis of MS in all cases. DISCUSSION/CONCLUSION: MS is a rare malignancy that shows pronounced clinical, morphologic, immunophenotypic, and genetic heterogeneity that often overlaps with other neoplastic and non-neoplastic entities. Features including the presence of classic myeloblasts, promonocytes, monoblasts, nucleated red blood cells, left-shifted granulocytes, cytoplasmic granules, and pseudopods are helpful hints in cytological preparations. In the modern era where pathologists are increasingly expected to do extensive diagnostic, molecular, and therapeutic biomarker testing on tissue that is historically diminishing in size, ROSE is a highly effective tool to ensure effective triage of MS aiding in an accurate, timely, and complete diagnosis.

Fine-needle aspiration of intramammary lymph nodes: a clinical, radiographic, and cytologic review.

INTRODUCTION: Recognizing and sampling intramammary lymph nodes (IMLNs) is important in the clinical management of patients with breast carcinomas. We undertook a retrospective study to evaluate the clinical utility of fine-needle aspiration (FNA) in assessing IMLNs. MATERIALS AND METHODS: Our pathology database was searched for all IMLN FNA cytology cases from January 2005 to December 2021. The cytologic findings, radiographic features, and clinical data were reviewed. RESULTS: A total of 149 cases were identified. Eighteen of 149 (12%) patients had synchronous breast tumors, including 13 invasive ductal carcinomas (IDCs), 1 ductal carcinoma in situ (DCIS), and 4 fibroadenomas. Among patients with synchronous IDCs, FNA of IMLNs was positive for metastatic carcinoma in 4 of 13 (30.7%) cases. The 4 patients with positive IMLNs all received mastectomies. Fifteen of 149 (10.7%) patients had a prior history of breast tumors, including 9 IDCs, 4 DCISs, 1 lobular carcinoma in situ (LCIS), and 1 fibroadenoma. Two of 149 (1.3%) patients had a prior history of lymphoma. In the patients with prior history of IDC, DCIS, LCIS, lymphomas and fibroadenomas, IMLN FNAs were all negative for malignancy. Two of 149 cases (1.3%) showed granulomatous lymphadenitis. The remaining 112 cases had negative IMLN FNAs and no significant clinical or pathological findings. CONCLUSIONS: Our study showed that IMLNs are commonly associated with synchronous/metachronous breast tumors (33 of 149, 22.1%). The incidence of positive IMLN FNA in patients with synchronous invasive breast carcinoma was 30.7% (4 of 13). FNA of IMLNs in conjunction with clinical presentation and radiologic findings allows triage of patients for appropriate clinical management and avoids additional unnecessary surgical procedures.

Growth improvement of wheat (Triticum aestivum) and zinc biofortification using potent zinc-solubilizing bacteria.

Zinc (Zn) is an indispensable element for proper plant growth. A sizeable proportion of the inorganic Zn that is added to soil undergoes a transformation into an insoluble form. Zinc-solubilizing bacteria (ZSB) have the potential to transform the insoluble Zn into plant-accessible forms and are thus promising alternatives for Zn supplementation. The current research was aimed at investigating the Zn solubilization potential of indigenous bacterial strains and to evaluate their impact on wheat growth and Zn biofortification. A number of experiments were conducted at the National Agriculture Research Center (NARC), Islamabad, during 2020-21. A total of 69 strains were assessed for their Zn-solubilizing ability against two insoluble Zn sources (ZnO and ZnCO3) using plate assay techniques. During the qualitative assay, the solubilization index and solubilization efficiency were measured. The qualitatively selected Zn-solubilizing bacterial strains were further tested quantitatively using broth culture for Zn and phosphorus (P) solubility. Tricalcium phosphate was used as insoluble source of P. The results showed that broth culture pH was negatively correlated with Zn solubilization, i.e., ZnO (r2 = 0.88) and ZnCO3 (r2 = 0.96). Ten novel promising strains, i.e., Pantoea sp. NCCP-525, Klebsiella sp. NCCP-607, Brevibacterium sp. NCCP-622, Klebsiella sp. NCCP-623, Acinetobacter sp. NCCP-644, Alcaligenes sp. NCCP-650, Citrobacter sp. NCCP-668, Exiguobacterium sp. NCCP-673, Raoultella sp. NCCP-675, and Acinetobacter sp. NCCP-680, were selected from the ecology of Pakistan for further experimentation on wheat crop based on plant growth-promoting rhizobacteria (PGPR) traits, i.e., solubilization of Zn and P in addition to being positive for nifH and acdS genes. Before evaluating the bacterial strains for plant growth potential, a control experiment was also conducted to determine the highest critical Zn level from ZnO to wheat growth using different Zn levels (0.1, 0.05, 0.01, 0.005, and 0.001% Zn) against two wheat varieties (Wadaan-17 and Zincol-16) in sand culture under glasshouse conditions. Zinc-free Hoagland nutrients solution was used to irrigate the wheat plants. As a result, 50 mg kg-1 of Zn from ZnO was identified as the highest critical level for wheat growth. Using the critical level (50 mg kg-1 of Zn), the selected ZSB strains were inoculated alone and in consortium to the seed of wheat, with and without the use of ZnO, in sterilized sand culture. The ZSB inoculation in consortium without ZnO resulted in improved shoot length (14%), shoot fresh weight (34%), and shoot dry weight (37%); with ZnO root length (116%), it saw root fresh weight (435%), root dry weight (435%), and Zn content in the shoot (1177%) as compared to the control. Wadaan-17 performed better on growth attributes, while Zincol-16 had 5% more shoot Zn concentration. The present study concluded that the selected bacterial strains show the potential to act as ZSB and are highly efficient bio-inoculants to combat Zn deficiency, and the inoculation of these strains in consortium performed better in terms of growth and Zn solubility for wheat as compared to individual inoculation. The study further concluded that 50 mg kg-1 Zn from ZnO had no negative impact on wheat growth; however, higher concentrations hampered wheat growth.

Earlier Return to Sports, Reduced Donor-Site Morbidity with Doubled Peroneus Longus Versus Quadrupled Hamstring Tendon Autograft in ACL Reconstruction.

Background: Graft choice for anterior cruciate ligament reconstruction (ACLR) has been evolving. The peroneus longus tendon (PLT) has been seen as a suitable choice for ACLR, providing comparable results to those of hamstring tendon (HT) autograft, but its clinical relevance in terms of return to sports, to our knowledge, has not been studied. Methods: Two hundred and thirty-two patients who sustained an isolated ACL injury were enrolled and underwent ACLR using doubled PLT autograft or quadrupled HT autograft; 158 were followed for 24 months. Functional scores (International Knee Documentation Committee [IKDC] and Tegner-Lysholm scores) were assessed preoperatively and at 3,6, 12, and 24 months postoperatively. Graft diameter and graft harvesting time were measured intraoperatively. Donor-site morbidity was evaluated using subjective evaluation. Time to return to sports in both groups was compared. Results: The mean diameter of PLT autograft was significantly larger than that of HT autograft, and the mean graft-harvesting time was less (p < 0.001). Patients in the PLT group returned to sports a mean of 34 days earlier than those in the HT group (p < 0.001) and had a lower rate of donor-site morbidity and, at 6 months, better patient-reported outcomes at the knee (p < 0.001). There were no significant differences between the groups in the rate of graft rupture or in IKDC and Tegner-Lysholm scores at the 24-month follow-up. Conclusions: PLT is a suitable autograft for ACLR in terms of graft diameter and graft-harvesting time and may offer athletes an earlier return to sports related to better outcomes at 6 months of follow-up. HT autograft was associated with increased thigh weakness. Both grafts, however, performed similarly at 24 months postoperatively. Level of Evidence: Therapeutic Level II. See Instructions for Authors for a complete description of levels of evidence.

Detection of Corynebacterium kroppenstedtii in Granulomatous Lobular Mastitis Using Real-Time Polymerase Chain Reaction and Sanger Sequencing on Formalin-Fixed, Paraffin-Embedded Tissues.

CONTEXT.­: Associations between granulomatous lobular mastitis (GLM) and Corynebacterium kroppenstedtii have been reported since 2002, but large-scale studies to assess the actual prevalence of this bacterium in GLM have not been performed. OBJECTIVE.­: To assess the prevalence of C kroppenstedtii in GLM using real-time polymerase chain reaction and Sanger sequencing. DESIGN.­: We analyzed formalin-fixed, paraffin-embedded tissues from 67 cases of GLM by sequential DNA amplification and sequencing to assess the rate of C kroppenstedtii detection in GLM. A retrospective analysis including patient demographics, history of pregnancy and lactation, clinical signs and symptoms, radiographic findings, histologic pattern, Gram stain results, and microbial cultures was performed on 67 cases of GLM. In addition, 10 cases of nongranulomatous breast abscess were included as controls. RESULTS.­: C kroppenstedtii 16S rRNA SYBR real-time polymerase chain reaction was positive on formalin-fixed, paraffin-embedded tissues from 46 of 67 (68.7%) GLM cases, while all control cases were negative. Among the positive cases, the majority showed features of cystic neutrophilic granulomatous mastitis. CONCLUSIONS.­: C kroppenstedtii was highly prevalent in GLM cases and was not found to be associated with nongranulomatous breast abscess in our study (P < .001).

SGLT2 Inhibitors in Type 2 Diabetes Mellitus and Heart Failure-A Concise Review.

The incidence of both diabetes mellitus type 2 and heart failure is rapidly growing, and the diseases often coexist. Sodium-glucose co-transporter 2 inhibitors (SGLT2i) are a new antidiabetic drug class that mediates epithelial glucose transport at the renal proximal tubules, inhibiting glucose absorption-resulting in glycosuria-and therefore improving glycemic control. Recent trials have proven that SGLT2i also improve cardiovascular and renal outcomes, including reduced cardiovascular mortality and fewer hospitalizations for heart failure. Reduced preload and afterload, improved vascular function, and changes in tissue sodium and calcium handling may also play a role. The expected paradigm shift in treatment strategies was reflected in the most recent 2021 guidelines published by the European Society of Cardiology, recommending dapagliflozin and empagliflozin as first-line treatment for heart failure patients with reduced ejection fraction. Moreover, the recent results of the EMPEROR-Preserved trial regarding empagliflozin give us hope that there is finally an effective treatment for patients with heart failure with preserved ejection fraction. This review aims to assess the efficacy and safety of these new anti-glycemic oral agents in the management of diabetic and heart failure patients.

Bone marrow-confined IL-6 signaling mediates the progression of myelodysplastic syndromes to acute myeloid leukemia.

Myelodysplastic syndromes (MDS) are age-related myeloid neoplasms with increased risk of progression to acute myeloid leukemia (AML). The mechanisms of transformation of MDS to AML are poorly understood, especially in relation to the aging microenvironment. We previously established an mDia1/miR-146a double knockout (DKO) mouse model phenocopying MDS. These mice develop age-related pancytopenia with oversecretion of proinflammatory cytokines. Here, we found that most of the DKO mice underwent leukemic transformation at 12-14 months of age. These mice showed myeloblast replacement of fibrotic bone marrow and widespread leukemic infiltration. Strikingly, depletion of IL-6 in these mice largely rescued the leukemic transformation and markedly extended survival. Single-cell RNA sequencing analyses revealed that DKO leukemic mice had increased monocytic blasts that were reduced with IL-6 knockout. We further revealed that the levels of surface and soluble IL-6 receptor (IL-6R) in the bone marrow were significantly increased in high-risk MDS patients. Similarly, IL-6R was also highly expressed in older DKO mice. Blocking of IL-6 signaling significantly ameliorated AML progression in the DKO model and clonogenicity of CD34-positive cells from MDS patients. Our study establishes a mouse model of progression of age-related MDS to AML and indicates the clinical significance of targeting IL-6 signaling in treating high-risk MDS.

Relapsing polychondritis: state-of-the-art review with three case presentations.

BACKGROUND: Relapsing polychondritis (RPC) is a complex immune-mediated systemic disease affecting cartilaginous tissue and proteoglycan-rich organs. The most common and earliest clinical features are intermittent inflammation involving the auricular and nasal regions, although all cartilage types can be potentially affected. The life-threatening effects of rpc involve the tracheobronchial tree and cardiac connective components. Rpc is difficult to identify among other autoimmune comorbidities; diagnosis is usually delayed and based on nonspecific clinical symptoms with limited laboratory aid and investigations. Medications can vary, from steroids, immunosuppressants, and biologics, including anti-tnf alpha antagonist drugs. METHOD: Information on updated etiology, clinical symptoms, diagnosis, and treatment of rpc has been obtained via extensive research of electronic literature published between 1976 and 2019 using PubMed and medline databases. English was the language of use. Search inputs included 'relapsing polychondritis,' 'polychondritis,' 'relapsing polychondritis symptoms,' and 'treatment of relapsing polychondritis.' Published articles in English that outlined and reported rpc's clinical manifestations and treatment ultimately met the inclusion criteria. Articles that failed to report the above and reported on other cartilaginous diseases met the exclusion criteria. RESULT: Utilizing an extensive overview of work undertaken in critical areas of RPC research, this review intends to further explore and educate the approach to this disease in all dimensions from pathophysiology, diagnosis, and management. CONCLUSION: RPC is a rare multi-systemic autoimmune disease and possibly fatal. The management remains empiric and is identified based on the severity of the disease per case. The optimal way to advance is to continue sharing data on RPC from reference centers; furthermore, clinical trials in randomized control groups must provide evidence-based treatment and management. Acquiring such information will refine the current knowledge of RPC, which will improve not only treatment but also diagnostic methods, including imaging and biological markers.

Gray Platelet Syndrome Presenting With Pancytopenia, Splenomegaly, and Bone Marrow Fibrosis.

OBJECTIVES: Gray platelet syndrome (GPS) is a rare platelet storage pool disorder associated with a marked decrease or absence of platelet α-granules and their contents. It is characterized clinically by mild to moderate bleeding; moderate macrothrombocytopenia with large, agranular platelets; splenomegaly; and bone marrow fibrosis. Electron microscopy confirms markedly reduced or absent α-granules in platelets and megakaryocytes. The classic description of GPS is caused by homozygous mutations in NBEAL2 (neurobeachinlike 2). METHODS: A 28-year-old Hispanic man with a history of easy bruising and occasional episodes of epistaxis sought treatment for pancytopenia and splenomegaly. Peripheral blood smear and bone marrow analysis, electron microscopy, and next-generation sequencing were performed. RESULTS: Large and agranular platelets were present in the peripheral blood. There was bone marrow fibrosis. Electron microscopy of the platelets showed absence of α-granules. Next-generation sequencing revealed a germline apparently homozygous nonsense variant in the NBEAL2 gene: c.5674C>T, p.Gln1892X (p.Q1829X). CONCLUSIONS: The differential diagnosis of GPS includes a myeloid neoplasm such as myelodysplastic syndrome with bone marrow fibrosis. The availability of diagnostic genetic panels for hereditable platelet disorders can assist in the recognition of GPS and other platelet disorders. We also describe a previously unreported pathogenic germline homozygous nonsense variant in the NBEAL2 gene: c.5674C>T, p.Gln1892X (p.Q1829X) in a patient with GPS.

Neonatal calf diarrhea: A potent reservoir of multi-drug resistant bacteria, environmental contamination and public health hazard in Pakistan.

Though emergence of multi-drug resistant bacteria in the environment is a demonstrated worldwide phenomenon, limited research is reported about the prevalence of resistant bacteria in fecal ecology of neonatal calf diarrhea (NCD) animals in Pakistan. The present study aimed to identify and assess the prevalence of bacterial pathogens and their resistance potential in the fecal ecology of NCD diseased animals of Pakistan. The presence of antibiotic resistance genes (blaTEM, blaNDM-1, blaCTX-M, qnrS) was also investigated. A total of 51 bacterial isolates were recovered from feces of young diarrheic animals (n = 11), collected from 7 cities of Pakistan and identified on the basis of 16S rRNA gene sequence and phylogenetic analysis. Selected isolates were subjected to antimicrobial susceptibility by disc diffusion method while polymerase chain reaction (PCR) was used to characterize the blaTEM, blaNDM-1, blaCTX-M, qnrS and mcr-1 antibiotic resistance genes. Based on the 16S rRNA gene sequences (Accession numbers: LC488898 to LC488948), all isolates were identified that belonged to seventeen genera with the highest prevalence rate for phylum Proteobacteria and genus Bacillus (23%). Antibiotic susceptibility explained the prevalence of resistance in isolates ciprofloxacin (100%), ampicillin (100%), sulfamethoxazole-trimethoprim (85%), tetracycline (75%), amoxicillin (55%), ofloxacin (50%), ceftazidime (45%), amoxicillin/clavulanic acid (45%), levofloxacin (30%), cefpodoxime (25%), cefotaxime (25%), cefotaxime/clavulanic acid (20%), and imipenem (10%). MICs demonstrated that almost 90% isolates were multi-drug resistant (against at least three antibiotics), specially against ciprofloxacin, and tetracycline with the highest resistance levels for Shigella sp. (NCCP-421) (MIC-CIP up to 75 µg mL-1) and Escherichia sp. (NCCP-432) (MIC-TET up to 250 µg mL-1). PCR-assisted detection of antibiotic resistance genes showed that 54% isolates were positive for blaTEM gene, 7% isolates were positive for blaCTX-M gene, 23% isolates were positive for each of qnrS and mcr-1 genes, 23% isolates were co-positive in combinations of qnrS and mcr-1 genes and blaTEM and mcr-1 genes, whereas none of the isolate showed presence of blaNDM-1 gene.

Accuracy of Intraoperative Frozen Section in Detection of Acute Invasive Fungal Rhinosinusitis.

CONTEXT.­: Acute invasive fungal rhinosinusitis (AIFRS) is an aggressive form of fungal sinusitis, which remains a significant cause of morbidity and mortality. Early diagnosis and intervention are keys to improving patient outcomes. Intraoperative consultation has shown promise in facilitating early surgical intervention, but the accuracy of frozen section has not been clarified in this setting. OBJECTIVES.­: To assess the accuracy of frozen-section diagnosis in patients with clinically suspected AIFRS. DESIGN.­: All cases of clinically suspected AIFRS during a 10-year period (2009-2019) were retrospectively reviewed. The frozen-section results were compared with the final permanent sections as well as the tissue fungal culture results, following which the accuracy of frozen section was determined. RESULTS.­: Forty-eight patients with 133 frozen-section evaluations for AIFRS were included in the study. Thirty of 48 patients and 61 of 133 specimens were positive for AIFRS on final pathology. Of 30 positive patients, 27 (90%) had at least 1 specimen diagnosed as positive during intraoperative consultation; among the 61 positive specimens, 54 (88.5%) were diagnosed as positive during intraoperative consultation. Of 72 negative specimens, all were interpreted as negative on frozen section. Thus, frozen sections had a sensitivity of 88.5% (95% CI, 0.78-0.97), specificity of 100% (95% CI, 0.94-1), positive predictive value of 100% (95% CI, 0.92-1), and negative predictive value of 90.6% (95% CI, 0.82-0.97). CONCLUSIONS.­: This study represents the largest series assessing the diagnostic accuracy of frozen section analysis in AIFRS. These findings are useful in frozen section-informed intraoperative decision making.

Draft genome sequence of a multidrug-resistant novel candidate Pseudomonas sp. NCCP-436T isolated from faeces of a bovine host in Pakistan.

OBJECTIVES: Here we describe the first draft genome analysis of a CRISPR-carrying, multidrug-resistant, candidate novel Pseudomonas sp. NCCP-436T isolated from faeces of a neonatal diarrhoeic calf. METHODS: The genome of strain NCCP-436T was sequenced using an Illumina NovaSeq PE150 platform and analysed using various bioinformatic tools. The virulence factors and resistome were identified using PATRIC and CARD servers, while CGView Server was used to construct a circular genome map. Antimicrobial susceptibility was determined by the disk diffusion technique. RESULTS: The draft genome of strain NCCP-436T contains 43 contigs with a total genome size of 3,683,517 bp (61.4% GC content). There are 3,452 predicted genes, including 60 tRNAs, 7 rRNAs and 12 sRNAs. CRISPR analysis revealed two CRISPR arrays with lengths of 1103 bp and 867 bp. Strain NCCP-436T was highly resistant to fluoroquinolone, ß-lactam, cephalosporin, aminoglycoside, penicillin, rifamycin, macrolide, glycopeptide, trimethoprim/sulfonamide and tetracycline antibiotic classes. Additionally, 22 antibiotic resistance genes, 313 virulence genes and 253 pathogen-host interactor genes were predicted. Comparison of the average nucleotide identity and digital DNA-DNA hybridisation values with the closely-related strain Pseudomonas khazarica (TBZ2) was found to be 82.08% and 34.90%, respectively, illustrating strain NCCP-436T as a potentially new species of Pseudomonas. CONCLUSION: Substantial number of antibiotic resistance and virulence genes and homology with human pathogens were predicted, exposing the pathogenic and zoonotic potential of strain NCCP-436T to public health. These findings may be used to better understand the genomic epidemiological features and drug resistance mechanisms of pathogenic Pseudomonas spp. in Pakistan.