bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evolution of ceftazidime–avibactam resistance driven by mutations in double-copy
bla
KPC-2
to
bla
KPC-189
during treatment of ST11 carbapenem-resistant
Klebsiella pneumoniae
Authors: Zhang Xiaofan , Xie Yinrong , Zhang Ying , Lei Tailong , Zhou Longjie , Yao Jiayao , Liu Lin , Liu Haiyang , He Jintao , Yu Yunsong , Tu Yuexing , Li Xi 
Published: September 2024

ABSTRACT

Klebsiella pneumoniae
carbapenemase (KPC) variants can contribute to resistance to ceftazidime–avibactam (CZA) in
Klebsiella pneumoniae
(KP). However, two-copy KPC variant-mediated resistance to CZA has rarely been reported to date. Here, we aimed to clarify the evolutionary trajectory of CZA resistance driven by mutations in double-copy
bla
KPC-2
to
bla
KPC-189
carried by the tandem core structure (IS
Kpn6-bla
KPC
-IS
Kpn27-tnpR
-IS
26
) during treatment of ST11 carbapenem-resistant
K. pneumoniae
(CRKP). The CZA-resistant KP strain carried double-copy
bla
KPC-189
, a variant with alanine–threonine and aspartate–tyrosine substitutions at Ambler amino acid positions 172 (A172T) and 179 (D179Y) of
bla
KPC-2
. Clone experiments confirmed that, compared with that of the wild-type
bla
KPC-2
clone strain, the minimum inhibitory concentration of CZA increased 16-fold in the
bla
KPC-189
-mutant strain. Furthermore, protein structure analysis revealed the A172T and D179Y mutations of
bla
KPC-189
can have a direct effect on the binding affinity of CAZ and AVI for KPC. Sequence comparison revealed that
bla
KPC-189
was mutated in a double-copy format upon CZA exposure, which was carried by the IS
26
-mediated tandem core structure IS
Kpn27-bla
KPC
-IS
Kpn6
. This tandem core structure apparently evolves
in vivo
during infection, although not by self-transferring, and multiple IS
Kpn27-bla
KPC
-IS
Kpn6
copy numbers could mediate transferable CZA resistance upon mobilization. In addition, compared with the wild-type
bla
KPC-2
gene, the
bla
KPC-189
gene had no fitness cost. In summary, our study highlighted the emergence of CZA-resistant
bla
KPC-189
variants in the ST11 clone and the presence of a double-copy
bla
KPC-189
in the IncFII-type plasmid, which is carried by a tandem core structure (IS
26-
IS
Kpn6-bla
KPC-189
-IS
Kpn27-tnpR
-IS
26
).

IMPORTANCE

To date, ceftazidime–avibactam (CZA) resistance caused by double-copy
Klebsiella pneumoniae
carbapenemase (KPC) variants has not been elucidated. The multicopy forms of carbapenem resistance genes carried by the same plasmid are relatively rare in most carbapenem-resistant
Enterobacteriaceae
. In this study, we elucidate the evolutionary trajectory of CZA resistance in ST11 carbapenem-resistant
K. pneumoniae
harboring a double-copy blaKPC and provide new insights into the mechanisms of acquired resistance to CZA.
Histone deacetylase HDAC3 regulates ergosterol production for oxidative stress tolerance in the entomopathogenic and endophytic fungus
Metarhizium robertsii
Authors: Liu Shuxing , Wang Xinmiao , Tang Xingyuan , Fang Weiguo 
Published: September 2024

ABSTRACT

Oxidative stress is encountered by fungi in almost all niches, resulting in fungal degeneration or even death. Fungal tolerance to oxidative stress has been extensively studied, but the current understanding of the mechanisms regulating oxidative stress tolerance in fungi remains limited. The entomopathogenic and endophytic fungus
Metarhizium robertsii
encounters oxidative stress when it infects insects and develops a symbiotic relationship with plants, and we found that host reactive oxygen species (ROSs) greatly limited fungal growth in both insects and plants. We identified a histone H3 deacetylase (HDAC3) that catalyzed the deacetylation of lysine 56 of histone H3. Deleting
Hdac3
significantly reduced the tolerance of
M. robertsii
to oxidative stress from insects and plants, thereby decreasing fungal ability to colonize the insect hemocoel and plant roots. HDAC3 achieved this by regulating the expression of three genes in the ergosterol biosynthesis pathway, which includes the lanosterol synthase gene
Las1
. The deletion of
Hdac3
or
Las1
reduced the ergosterol content and impaired cell membrane integrity. This resulted in an increase in ROS accumulation in fungal cells that were thus more sensitive to oxidative stress. We further showed that HDAC3 regulated the expression of the three ergosterol biosynthesis genes in an indirect manner. Our work significantly advances insights into the epigenetic regulation of oxidative stress tolerance and the interactions between
M. robertsii
and its plant and insect hosts.

IMPORTANCE

Oxidative stress is a common challenge encountered by fungi that have evolved sophisticated mechanisms underlying tolerance to this stress. Although fungal tolerance to oxidative stress has been extensively investigated, the current understanding of the mechanisms for fungi to regulate oxidative stress tolerance remains limited. In the model entomopathogenic and plant symbiotic fungus
Metarhizium robertsii
, we found that the histone H3 deacetylase HDAC3 regulates the production of ergosterol, an essential cell membrane component. This maintains the cell membrane integrity to resist the oxidative stress derived from the insect and plant hosts for successful infection of insects and development of symbiotic associates with plants. Our work provides significant insights into the regulation of oxidative stress tolerance in
M. robertsii
and its interactions with insects and plants.
First detection of VEB-1 extended-spectrum β-lactamase-producing
Escherichia coli
clinical isolate in Japan
Authors: Shindou Junko , Hayashi Wataru , Kayama Shizuo , Yu Liansheng , Zuo Hui , Sugawara Yo , Sugai Motoyuki 
Published: September 2024

ABSTRACT

The extended-spectrum β-lactamase (ESBL) gene,
bla
VEB-1
, was identified for the first time in an
Escherichia coli
clinical isolate, JARB-RN-0061, from blood cultures in a Japanese general hospital in 2021. The isolate exhibited high resistance to broad-spectrum cephalosporins, including ceftazidime (MIC >128 mg/L) and cefepime (MIC = 16 mg/L).
bla
VEB-1
was identified during whole-genome sequencing and characterization of the isolate. JARB-RN-0061 belonged to the B2-O2:K1:H7-ST95-
fimH
41 lineage and was classified as presumptive extraintestinal pathogenic
E. coli
(ExPEC) and uropathogenic
E. coli
(UPEC). Moreover, the strain harbored multiple virulence genes on the chromosome. The Col156/IncFIB(AP001918)/IncFII(29)-type plasmid (114,216 bp), with
clbB
and
tcpC
genes involved in bacteremia, was unique to the
fimH
41 subclone. The
bla
VEB-1
gene was located on a non-typeable and non-conjugative plasmid, pJARB-RN-0061_VEB-1 (17,093 bp). It was embedded in the class 1 integron In1883-like, with multidrug resistance gene cassettes for
aacA4
,
aadB
,
cmlA5
,
qnrVC4
, and
dfrA14
. Notably, comparative analysis of the complete sequence of plasmid pJARB-RN-0061_VEB-1 revealed that it was highly homologous to the
bla
VEB-1
-harboring plasmid, pMS2H7VEB-1 (100% coverage and 99.99% identity), except for the Tn3 family transposon (4,931 bp) and the plasmid pRHBSTW-00138_5 (97% coverage and 100% identity) harbored by
Klebsiella quasipneumoniae
subsp.
similipneumoniae
strains from hospital sewage in Japan and wastewater influent in the United Kingdom, respectively. The emergence of a human pathogenic
E. coli
clinical isolate with the
bla
VEB-1
-carrying plasmid in the B2-ST95 worldwide pandemic lineage, characterized by the virulence potential of ExPEC or UPEC but a low prevalence of antimicrobial resistance, would raise public health concerns.

IMPORTANCE

ESBLs are plasmid-mediated enzymes that confer resistance to clinically significant antimicrobial agents, such as broad-spectrum cephalosporins. Recently, the rapid spread of CTX-M-type ESBL-producing
E. coli
has become a global issue, including in Japan, where ESBL production in human pathogenic
E. coli
, such as the ExPEC and UPEC lineages, which typically harbor several virulence genes, is a severe public health concern. To date, VEB (Vietnamese extended-spectrum β-lactamase) producers have been found only in hospital wastewater and rivers in Japan. Thus, we describe the first detection of a very rare human-derived blaVEB-1 gene in the
E. coli
B2-ST95 pandemic clonal lineage that is highly associated with ExPEC and UPEC in a Japanese clinical setting. Furthermore, we characterized the genomic features of plasmids harboring the class 1 integron-borne blaVEB-1. Our findings highlight the significance of closely monitoring ESBL-producing
E. coli
isolates to prevent the potential dissemination of this resistance determinant in Japan.
Ultrahigh-pressure to high-pressure eclogite in Cuban ophiolitic mélange reveals proto-Caribbean spreading ridge subduction
Authors: Rui Hui-Chao , Duan Wen-Yong , Yang Jing-Sui , Maresch Walter V. , Schertl Hans-Peter , Cai Peng-Jie , Llanes-Castro Angélica I. , He Hong-Ping 
Published: September 2024

Abstract
Proto-Caribbean oceanic crust produced during ocean-floor spreading between diverging North and South American plates was subsequently subducted beneath the Caribbean plate. However, the timing and spatial configuration of proto-Caribbean spreading ridge subduction remain subjects of debate. High-pressure (HP) basaltic metamorphic rocks, representing relics of the subducted proto-Caribbean oceanic crust, commonly occur in Cuban ophiolitic mélanges. In this study, an integrated set of petrological, geochemical, and geochronological data is presented for eclogite from the Las Villas mélange, central Cuba. The typical geochemical signature of mid-ocean-ridge basalt (MORB) indicates that the protolith of eclogite formed at the proto-Caribbean spreading ridge. Based on pressure-temperature (P-T) estimates obtained by pseudosection analysis as well as Zr-in-rutile and Ti-in-zircon thermometry, the following P-T paths for representative samples can be derived: a prograde path from 24–25 kbar and 510–520 °C to peak conditions of 29–31 kbar and 525–575 °C, and a complex retrograde path initially following almost isothermal exhumation to 25–27 kbar, followed by near-isobaric heating to 610–640 °C before final exhumation. This is the first documentation of prograde oceanic ultrahigh-pressure (UHP) metamorphism in the northern Caribbean area. U-Pb dating of magmatic zircon with steep heavy rare earth element (HREE) patterns and negative Eu anomalies yielded a protolith age of 126.3 ± 0.7 Ma. In contrast, metamorphic zircon with flat HREE patterns and without an Eu anomaly yielded a weighted mean age of 118.6 ± 1.6 Ma. The short time interval of &gt;8 m.y. between MORB magmatism and UHP metamorphism suggests that the oceanic crust was subducted to great depth (~100 km) shortly after generation in an oceanic ridge, which provides robust evidence for subduction of the proto-Caribbean spreading ridge. Furthermore, this work demonstrates high potential to trace ancient spreading ridge subduction by joint petrological, geochemical, and geochronological study of oceanic eclogite.
Vascular mimicry and mosaic vessels in parathyroid tumours: a new diagnostic approach?
Authors: Falleni Monica , Dal Lago Matteo , Tosi Delfina , Ghilardi Giorgio , De Pasquale Loredana , Saibene Alberto M , Felisati Giovanni , Cozzolino Mario , Gianelli Umberto 
Published: September 2024

AimsEvaluation of ‘alternative’ vascularisation in human cancer is considered an important prognostic parameter; the 2022 WHO classification of parathyroid tumours despite progresses in clinical triaging of patients strongly emphasises new histopathological parameters to properly stratify these lesions. ‘Alternative’ and ‘classic’ vessels were here investigated for the first time in parathyroid tumours for their possible histopathological and clinical relevance during progression.MethodsUsing a double CD31/PAS staining, microvessel density (MVD, ‘classic’ CD31+ vessels), mosaic vessel density (MoVD, ‘alternative’ CD31+/−vessels) and vessel mimicry density (VMD, ‘alternative’ CD31−/PAS+ vessels) were evaluated in 4 normal parathyroid glands (N), 50 Adenomas (A), 35 Atypical Tumours (AT) and 10 Carcinomas (K).ResultsCompared with N, MVD significantly increased in A (p=0.012) and decreased in K (p=0.013) with vessel counts lower than in AT and A (p<0.001). MoVs and VMs, absent in normal tissue, were documented in non-benign parathyroid lesions (AT, K) (p<0.001), with MoVs and VMs most represented in AT and K, respectively (p<0.001), in peripheral growing areas. Vessel distribution was correlated to neoplastic progression (r=−0.541 MVD; r=+0.760 MoVD, r=+0.733 VMD), with MVD decrease in AT and K inversely related to MoVD and VMD increase (r=−0.503 and r=−0.456).Conclusions‘Alternative’ vessel identification in parathyroid tumours is crucial because it: (1) explains the paradox of non-angiogenic tumours, consisting in a new bloody non-endothelial vessel network and (2) helps pathologists to unmask worrisome lesions. Furthermore, detection of alternative vascular systems in human tumours might explain the limited success of antiangiogenic therapies and encourage new oncological studies.
Managing low-level HIV viraemia in antiretroviral therapy: a systematic review and meta-analysis
Authors: Zaçe Drieda , Rindi Lorenzo Vittorio , Compagno Mirko , Colagrossi Luna , Santoro Maria Mercedes , Andreoni Massimo , Perno Carlo Federico , Sarmati Loredana 
Published: September 2024

Objective
HIV-1 management has advanced significantly with antiretroviral therapy (ART), yet challenges persist, including low-level HIV-1 viraemia (LLV). LLV presents a complex scenario, with varied definitions in the literature, reflecting uncertainties in its clinical interpretation. Questions arise regarding the underlying mechanisms of LLV, whether it signifies ongoing viral replication or stems from other factors. This study aimed to systematically review strategies for LLV management, providing insights into optimal clinical approaches.

Methods
MEDLINE, EMBASE, Cochrane Library, Web of Science and Canadian Agency for Drugs and Technologies in Health were searched for relevant literature on LLV management. We included studies published between 2004 and 2024, assessing interventions such as ART modification, genotypic resistance testing, adherence assessment, performing therapeutic drug monitoring, testing for chronic coinfections and assessing the viral reservoir via HIV DNA quantification. Meta-analyses were conducted where feasible.

Results
The systematic review identified 48 eligible records. Findings indicated limited evidence supporting the effectiveness of ART regimen modification in achieving virological suppression among individuals with LLV. However, studies assessing genotypic resistance testing revealed a significant association between resistance-associated mutations and virological suppression during LLV. Adherence to ART emerged as a critical determinant of treatment efficacy, with interventions showing promise in achieving viral suppression. The clinical utility of therapeutic drug monitoring in managing LLV remained inconclusive. Gaps in the literature were identified regarding follow-up scheduling, managing concurrent chronic infections and assessing inflammatory markers in LLV management.

Conclusions
While ART modification may not consistently achieve virological suppression, genotypic resistance testing may offer insights into treatment outcomes. Adherence to ART emerged as a crucial factor, necessitating tailored interventions. However, further research is needed to elucidate the clinical utility of therapeutic drug monitoring and other management strategies. The study highlights the importance of ongoing research to refine therapeutic approaches and improve patient outcomes in LLV management.

PROSPERO registration number
CRD42024511492.
Availability and Price Variations of Commonly Used Cardiovascular Medicines at Community and Hospital Pharmacies in Gondar Town, Northwest Ethiopia
Authors: Limenh Liknaw Workie , Tessema Tewodros Ayalew , Sendekie Ashenafi Kibret , Simegn Wudneh , Ayenew Wondim , Mitku Melese Legesse , Anagaw Yeniewa Kerie , Kasahun Asmamaw Emagn 
Published: September 2024

Background: Access to cardiovascular medications is severely hampered by their unavailability and high cost, particularly for the poorest households in developing nations. The availability and price range of cardiovascular medications are significantly limited in both hospital and community pharmacies.Objectives: The aim of this study is to assess the availability and price variations of commonly used cardiovascular medicines in hospital pharmacies in Gondar Town, northwest Ethiopia.Methods: From July 13 to August 6, 2022, a mixed cross‐sectional and simulated client survey was carried out at two hospital and 13 community pharmacies in Gondar Town. The analysis and data entry were performed using SPSS Version 25 and EpiData Version 4.2, respectively. The availability and pricing variations of the medications are given as percentages. The significance was examined using paired t tests.Results: On average, community retail pharmacies offered 33.22% of CVD drugs. Aspirin (81 mg), amlodipine (5 mg), atorvastatin (20 mg), and hydrochlorothiazide (25 mg) were the most readily available drugs in community pharmacies. Overall, 28.00% of the hospital pharmacies had available CVD medicines during the course of our analysis. The average cost for the 25 CVD medications in hospital pharmacies was $0.699, with a standard deviation (SD) of 1.513, which was less than the cost at community pharmacies ($2.741 with an SD of 6.015) (p = 0.045).Conclusion: CVD medications were more available in community pharmacies than in hospital pharmacies, although there were fewer CVD medications available than recommended by the WHO/HAI (80%) in both hospital and community pharmacies. There was a statistically significant difference between the two prices. Compared to that at hospital pharmacies, the mean price at community pharmacies was greater.
CD24-Targeted NIR-II Fluorescence Imaging Enables Early Detection of Colorectal Neoplasia
Authors: Guo Xiaoyong , Luo Shuangling , Wang Xiaofeng , Cui Yingying , Li Miaomiao , Zhang Zeyu , Fu Lidan , Cao Caiguang , Shi Xiaojing , Liu Haifeng , Qu Yawei , Gao Xiangyu , Hu Zhenhua , Tian Jie 
Published: September 2024

Abstract
Colorectal cancer continues to be a major health issue even though screening methods have facilitated early detection. Despite the high sensitivity of white-light colonoscopy, it frequently overlooks invasive flat or depressed lesions, which can lead to the development of larger, advanced tumors. Fluorescence molecular imaging offers a promising approach for early tumor detection by targeting specific molecular characteristics of lesions. CD24 is upregulated during the adenoma-to-colorectal cancer transition, providing a potential target for fluorescence molecular imaging. In this study, we developed a second near-infrared window (NIR-II) fluorescent probe with a high affinity for CD24 and evaluated its efficacy and targeting ability in cellular models, murine models, and clinical samples of colorectal cancer. CD24 expression was elevated in 76% of adenomas and 80% of colorectal cancers. In a colitis-associated cancer mouse model, NIR-II imaging with the CD24-targeted probe achieved a significantly higher tumor-to-background ratio compared with conventional NIR-I imaging. The probe demonstrated exceptional sensitivity (92%) and specificity (92%) for detecting colorectal cancer, including small lesions less than 1 mm in size. This led to the identification of precancerous lesions missed by white-light detection and lesions missed by NIR-I imaging. Moreover, ex vivo human tissue incubation with the probe supported the potential for intraprocedural lesion identification via topical probe application during colonoscopy. In conclusion, this study successfully demonstrates the potential of CD24-targeted NIR-II imaging for identifying colorectal neoplasia, highlighting its significance for early colorectal cancer detection in the gastrointestinal tract.
Significance: Overexpression of CD24 in colorectal dysplasia provides the opportunity to use an NIR-II fluorescent probe targeting CD24 to detect colorectal neoplasms, including invisible lesions that are missed by white-light colonoscopy.
A B7-H3–Targeted CD28 Bispecific Antibody Enhances the Activity of Anti–PD-1 and CD3 T-cell Engager Immunotherapies
Authors: Moore Gregory L. , Zeng Veronica G. , Diaz Juan E. , Bonzon Christine , Avery Kendra N. , Rashid Rumana , Qi Jing , Nam Dong Hyun , Jacinto Jonathan , Dragovich Matthew A. , Kim Yoon Kyung , Balcazar Karen P. , Bakhit Charles G. , Eivazi Araz , Nguyen Hanh , Muchhal Umesh S. , Szymkowski David E. , Desjarlais John R. , Hedvat Michael 
Published: September 2024

Abstract
T-cell activation is a multistep process requiring T-cell receptor engagement by peptide–MHC complexes (Signal 1) coupled with CD28-mediated costimulation (Signal 2). Tumors typically lack expression of CD28 ligands, so tumor-specific Signal 1 (e.g., neoepitope presentation) without costimulation may be ineffective or even induce T-cell anergy. We designed the bispecific antibody XmAb808 to co-engage the tumor-associated antigen B7-H3 with CD28 to promote T-cell costimulation within the tumor microenvironment. XmAb808 costimulation was measured by its ability to activate and expand T cells and enhance T cell–mediated cancer cell killing in cocultures of human peripheral blood mononuclear cells and cancer cells and in mice engrafted with human peripheral blood mononuclear cells and tumor xenografts. XmAb808 avidly bound cancer cells and stimulated IL2 and IFNγ secretion from T cells cocultured with cancer cells engineered to deliver Signal 1 to T cells via a surface-expressed anti-CD3 antibody. XmAb808 enhanced expression of the antiapoptotic factor Bcl-xL and CD25, promoting survival and IL2-dependent expansion of T cells coupled with increased T cell–mediated cytotoxicity in vitro. XmAb808 combined with an EpCAM×CD3 bispecific antibody to enhance target cell killing through IL2-dependent expansion of CD25+ T cells. This combination also suppressed pancreatic tumor xenograft growth in mice. Furthermore, XmAb808 combined with an anti–programmed cell death protein 1 antibody to suppress breast tumor xenograft growth in mice. XmAb808 as monotherapy and in combination with an anti–programmed cell death protein 1 antibody is currently in clinical development in patients with advanced solid tumors. Our results suggest that XmAb808 may also combine with tumor antigen–targeted anti-CD3 (Signal 1) T-cell engagers.
The London Regime’s Adaptability and Impact
Authors: Pacheco Castillo José M , Seybold Dorothee , Santillo David 
Published: September 2024

Abstract
The London Convention (LC), adopted in 1972, and its successor, the London Protocol (LP) of 1996, constitute key regulatory frameworks working in parallel, aimed at mitigating waste dumping at sea and safeguarding the marine environment. These treaties, which share the objective of controlling marine pollution, have also evolved to address emerging environmental issues such as marine geoengineering and carbon capture and storage within the context of the current triple planetary crisis. As the LC/LP regime marked its fiftieth anniversary in 2022, both achievements and ongoing challenges in combating pollution from dumping activities were acknowledged, emphasising the need for continual adaptation to protect the marine environment effectively. This article examines the adaptability and impact of the LC/LP regime over the past five decades, identifying strengths and proposing future steps to ensure the preservation of ocean health for future generations.