bluuebunny/crossref_metadata_2025_split huggingface dataset:
Engineering the thermal stability of a polyphosphate kinase by ancestral sequence reconstruction to expand the temperature boundary for an industrially applicable ATP regeneration system
Authors: Li Zong-Lin , Sun Chuan-Qi , Qing Zhou-Lei , Li Zhi-Min , Liu Hong-Lai 
Published: January 2024

ABSTRACT

ATP-dependent energy-consuming enzymatic reactions are widely used in cell-free biocatalysis. However, the direct addition of large amounts of expensive ATP can greatly increase cost, and enzymatic production is often difficult to achieve as a result. Although a polyphosphate kinase (PPK)-polyphosphate-based ATP regeneration system has the potential to solve this challenge, the generally poor thermal stability of PPKs limits the widespread use of this method. In this paper, we evaluated the thermal stability of a PPK from
Sulfurovum lithotrophicum
(
Sl
PPK2). After directed evolution and computation-supported design, we found that
Sl
PPK2 is very recalcitrant and cannot acquire beneficial mutations. Inspired by the usually outstanding stability of ancestral enzymes, we reconstructed the ancestral sequence of the PPK family and used it as a guide to construct three heat-stable variants of
Sl
PPK2, of which the L35F/T144S variant has a half-life of more than 14 h at 60°C. Molecular dynamics simulations were performed on all enzymes to analyze the reasons for the increased thermal stability. The results showed that mutations at these two positions act synergistically from the interior and surface of the protein, leading to a more compact structure. Finally, the robustness of the L35F/T144S variant was verified in the synthesis of nucleotides at high temperature. In practice, the use of this high-temperature ATP regeneration system can effectively avoid byproduct accumulation. Our work extends the temperature boundary of ATP regeneration and has great potential for industrial applications.

IMPORTANCE

ATP regeneration is an important basic applied study in the field of cell-free biocatalysis. Polyphosphate kinase (PPK) is an enzyme tool widely used for energy regeneration during enzymatic reactions. However, the thermal stability of the PPKs reported to date that can efficiently regenerate ATP is usually poor, which greatly limits their application. In this study, the thermal stability of a difficult-to-engineer PPK from
Sulfurovum lithotrophicum
was improved, guided by an ancestral sequence reconstruction strategy. The optimal variant has a 4.5-fold longer half-life at 60°C than the wild-type enzyme, thus enabling the extension of the temperature boundary for ATP regeneration. The ability of this variant to regenerate ATP was well demonstrated during high-temperature enzymatic production of nucleotides.
On-site filtration of large sample volumes improves the detection of opportunistic pathogens in drinking water distribution systems
Authors: Hozalski Raymond M. , Zhao Xiaotian , Kim Taegyu , LaPara Timothy M. 
Published: January 2024

ABSTRACT

In this study, we compared conventional vacuum filtration of small volumes through disc membranes (effective sample volumes for potable water: 0.3–1.0 L) with filtration of high volumes using ultrafiltration (UF) modules (effective sample volumes for potable water: 10.6–84.5 L) for collecting bacterial biomass from raw, finished, and tap water at seven drinking water systems. Total bacteria,
Legionella
spp.,
Legionella pneumophila
,
Mycobacterium
spp., and
Mycobacterium avium
complex in these samples were enumerated using both conventional quantitative PCR (qPCR) and viability qPCR (using propidium monoazide). In addition, PCR-amplified gene fragments were sequenced for microbial community analysis. The frequency of detection (FOD) of
Legionella
spp. in finished and tap water samples was much greater using UF modules (83% and 77%, respectively) than disc filters (24% and 33%, respectively). The FODs for
Mycobacterium
spp. in raw, finished, and tap water samples were also consistently greater using UF modules than disc filters. Furthermore, the number of observed operational taxonomic units and diversity index values for finished and tap water samples were often substantially greater when using UF modules as compared to disc filters. Conventional and viability qPCR yielded similar results, suggesting that membrane-compromised cells represented a minor fraction of total bacterial biomass. In conclusion, our research demonstrates that large-volume filtration using UF modules improved the detection of opportunistic pathogens at the low concentrations typically found in public drinking water systems and that the majority of bacteria in these systems appear to be viable in spite of disinfection with free chlorine and/or chloramine.

IMPORTANCE

Opportunistic pathogens, such as
Legionella pneumophila
, are a growing public health concern. In this study, we compared sample collection and enumeration methods on raw, finished, and tap water at seven water systems throughout the State of Minnesota, USA. The results showed that on-site filtration of large water volumes (i.e., 500–1,000 L) using ultrafiltration membrane modules improved the frequency of detection of relatively rare organisms, including opportunistic pathogens, compared to the common approach of filtering about 1 L using disc membranes. Furthermore, results from viability quantitative PCR (qPCR) with propidium monoazide were similar to conventional qPCR, suggesting that membrane-compromised cells represent an insignificant fraction of microorganisms. Results from these ultrafiltration membrane modules should lead to a better understanding of the microbial ecology of drinking water distribution systems and their potential to inoculate premise plumbing systems with opportunistic pathogens where conditions are more favorable for their growth.
Successive accretions of future allochthonous terranes and multiple subduction zone jumps: Implications for Tethyan evolution
Authors: Yan Zhiyong , Chen Lin , Zuza Andrew V. , Meng Qingren 
Published: January 2024

Abstract
The accretion of future allochthonous terranes (e.g., microcontinents or oceanic plateaus) onto the southern margin of Asia occurred repeatedly during the evolution and closure of the Tethyan oceanic realm, but the specific geodynamic processes of this protracted convergence, successive accretion, and subduction zone initiation remain largely unknown. Here, we use numerical models to better understand the dynamics that govern multiple terrane accretions and the polarity of new subduction zone initiation. Our results show that the sediments surrounding the future terranes and the structural complexity of the overriding plate are important factors that affect accretion of multiple plates and guide subduction polarity. Wide (≥400 km) and buoyant terranes with sediments behind them and fast continental plate motions are favorable for multiple unidirectional subduction zone jumps, which are also referred to as subduction zone transference, and successive terrane-accretion events. The jumping times (~3–20 + m.y.) are mainly determined by the convergence rates and rheology of the overriding complex plate with preceding terrane collisions, which increase with slower convergence rates and/or a greater number of preceding terrane collisions. Our work provides new insights into the key geodynamic conditions governing multiple subduction zone jumps induced by successive accretion and discusses Tethyan evolution at a macro level. More than 50 m.y. after India-Asia collision, subduction has yet to initiate along the southern Indian plate, which may be the joint result of slower plate convergence and partitioned deformation across southern Asia.
Increase in magma supply to Sakurajima volcano’s (Japan) shallow magma chamber over the past 500 years
Authors: Huber Christian , Toramaru Atsushi 
Published: January 2024

Abstract
Constraining the magma supply to a subvolcanic reservoir is crucial to characterizing unrest and evaluating the potential for a forthcoming eruption. With the advent of GPS, tiltmeters, and satellite interferometry, it has been possible to infer changes in the supply rate of magma to shallow reservoirs over time scales of as much as decades. While these methods have impacted our ability to characterize volcanic unrest, they only probe magmatic activity over short time scales. Here, we constrain changes in magma supply rate at Sakurajima volcano (Kyushu, Japan) over the past five centuries. The combination of thermo-mechanical modeling, documented eruption history, and textural analyses of pumices allows us to tightly constrain the long-term rate of magma supply to the subvolcanic chamber through time. Specifically, we find that magma supply rate has increased by roughly an order of magnitude over the past 500 years and that the bubble content in the magma chamber has increased over time, explaining the changes in eruption volume between the Bunmei (ca. A.D. 1470), An-ei (ca. A.D. 1780), and Taisho (A.D. 1914) eruptions.
Does insulin pump therapy offer benefits for behaviour, mood, cognition and HbA1c in children and adolescents with type 1 diabetes? A randomised controlled trial with observational follow-up
Authors: O'Connell Michele A , Northam Elisabeth A , Brown Amy , Papoutsis Jennifer , Schuster Tibor , Skinner Timothy , Jenkins Alicia J , Ambler Geoffrey R , Cameron Fergus J 
Published: January 2024

AimsImproved behaviour, mood, cognition and HbA1c have been reported with short-term use of continuous subcutaneous insulin infusion (CSII) in youth with type 1 diabetes (T1D). We sought to re-examine these findings in a randomised controlled trial (RCT), with longitudinal follow-up.MethodsRCT of youth aged 7–15 years with T1D, at two tertiary paediatric centres. Participants were randomised to commence CSII or continue multiple daily injections (MDI). Behaviour, mood, cognition and HbA1c were assessed. Primary outcome was difference in parent-reported behaviour (BASC-2) at 4 months. After the 4-month RCT, MDI participants commenced CSII; outcomes were reassessed at +2 years.ResultsParticipating youth (n=101) were randomised to CSII (n=56) or MDI (n=45). Significant differences favouring CSII were found at 4 months in parent-reported behaviour problems (Cohen’s d 0.41 (95% CI 0.004 to 0.795); p=0.048) and HbA1c (mean (95% CI) difference: 7 (2.3 to 11.7) mmol/mol (0.6% (0.2 to 1.0%); p=0.001)). Improvements from baseline were documented in mood and cognitive outcomes in both study groups over the 4-month RCT; however, no between-group differences were evident at 4 months. Sixteen of 76 (21%) participants completing assessments at +2 years had discontinued CSII. In n=60 still using CSII, measurements of behaviour, mood and HbA1c were comparable to baseline.ConclusionsParent-reported behaviour problems and HbA1c, but not mood or neurocognitive outcomes, were clinically significantly lower with CSII, relative to MDI, after 4 months. Observational follow-up indicated no impact of treatment modality at +2 years, relative to baseline levels. Taken together, these data indicate that use of CSII alone does not comprehensively benefit neuropsychological outcomes in childhood T1D.
Pattern and prognostic factors for kidney function progression in survivors of children with acute kidney injury: a cohort study
Authors: Hui Wun Fung , Chan Vivian Pui Ying , Cheung Wing Lum , Ku Shu Wing , Hon Kam Lun 
Published: January 2024

ObjectiveTo examine the pattern of kidney function progression after acute kidney injury (AKI) and identify the associated risk factors.DesignA prospective cohort study was conducted from June 2020 to June 2021 on children aged 1 month to <18 years admitted to the paediatric intensive care unit (PICU). Acute kidney disease (AKD) was defined as AKI persisting from 7 to 90 days after diagnosis. The natural history and prognostic factors of kidney function progression were determined.ResultsAmong the 253 admissions with a median (IQR) age of 4.9 (9.7) years, the AKI and AKD incidence was 41.9% and 52.2% respectively. The incidence of estimated glomerular filtration rate (eGFR) <90 mL/min/1.73 m2was 6.7% at 90 days and 11.9% at latest follow-up. Severe and prolonged AKI and higher degree of nephrotoxic medication exposure were associated with AKD development. The severity and duration of AKI and AKD significantly predicted kidney function non-recovery. Children with both entities exhibited a higher peak-to-baseline serum creatinine level ratio at 90 days (1.6 vs 1.0, p<0.001), and a more pronounced decline in eGFR (21% vs 19%, p=0.028) during the follow-up period compared with those without AKI/AKD. They also had an increased risk of having eGFR <90 mL/min/1.73 m2at 90 days (HR 14.9 (95% CI 1.8 to 124.0)) and latest follow-up (HR 3.8 (95% CI 1.1 to 13.1)).ConclusionsAKI and AKD are prevalent among critically ill children and pose substantial risk for non-recovery of kidney function among PICU survivors. A structural follow-up visit for AKI survivors to monitor kidney function progression is advocated.
Neuroimaging changes in the pregeniculate visual pathway and chiasmal enlargement in Leber hereditary optic neuropathy
Authors: Xu Xintong , Zhou Huanfen , Sun Mingming , Li Yuyu , Chen Biyue , Chen Xiyun , Xu Quangang , Yu-Wai-Man Patrick , Wei Shihui 
Published: January 2024

PurposeTo describe the pattern of MRI changes in the pregeniculate visual pathway in Leber hereditary optic neuropathy (LHON).MethodThis retrospective observational study enrolled 60 patients with LHON between January 2015 and December 2021. The abnormal MRI features seen in the pregeniculate visual pathway were investigated, and then correlated with the causative mitochondrial DNA (mtDNA) mutation, the distribution of the MRI lesions and the duration of vision loss.ResultThe cohort included 48 (80%) males and 53 (88%) had bilateral vision loss. The median age of onset was 17.0 years (range 4.0–58.0). 28 (47%) patients had the m.11778G>A mutation. 34 (57%) patients had T2 hyperintensity (HS) in the pregeniculate visual pathway and 13 (22%) patients with chiasmal enlargement. 20 patients (71%) carrying the m.11778G>A mutation had T2 HS, significantly more than the 14 patients (44%) with T2 HS in the other LHON mutation groups (p=0.039). Furthermore, significantly more patients in the m.11778G>A group (16 patients (57%)) had T2 HS in optic chiasm (OCh)/optic tract (OTr) than the other LHON mutation groups (7 patients (22%), p=0.005). Optic chiasmal enlargement was more common in patients with vision loss duration <3 months compared with those ≥3 months (p=0.028).ConclusionT2 HS in the pregeniculate visual pathway is a frequent finding in LHON. Signal changes in the OCh/OTr and chiasmal enlargement, in particular within the first 3 months of visual loss, were more commonly seen in patients carrying the m.11778G>A mtDNA mutation, which may be of diagnostic significance.
Sri Lanka’s COVID-19 response and maintaining health services: implications for future pandemics
Authors: Rannan-Eliya Ravindra Prasan , Ghaffoor Azrah , Amarasinghe Sarasi , Nirmani Manage Dhanusha , Wijemunige Nilmini , Perera Sanjay , Samarage Sarath , Dalpatadu KCS , Wisidagama Niroshani , Fonseka Sachini , Kumara Nalin , Selvaratnam Ahthiga , Dananjana Thimani 
Published: January 2024

This study examines how Sri Lanka, a lower-middle income country, managed its COVID-19 response and maintained health services. It draws on an extensive document review, key informant interviews and a national survey of public experience and opinion to assess what Sri Lanka did, its effectiveness and why.Owing to a strong health system and luck, Sri Lanka stopped the first wave of COVID-19 infections, and it adopted a ‘Zero-COVID’ approach with the explicit goal of stopping outbreaks. This was initially effective. Outbreaks reduced healthcare use, but with minimal impact on health outcomes. But from end-2020, Sri Lanka switched its approach to tolerating transmission and mitigation. It took proactive actions to maintain healthcare access, and it pursued a COVID-19 vaccination effort that was successful in covering its adult population rapidly and with minimal disparities. Despite this, widespread transmission during 2021–2022 disrupted health services through the pressure on health facilities of patients with COVID-19 and infection of healthcare workers, and because COVID-19 anxiety discouraged patients from seeking healthcare. This led to substantial mortality and more than 30 000 excess deaths by 2022.We find that Sri Lanka abandoned its initially successful approach, because it failed to understand that its chosen strategy required symptomatic PCR testing in primary care. Failure to invest in testing was compounded by groupthink and a medical culture averse to testing.Sri Lanka’s experience confirms that strong public health capacities, robust healthcare systems and intersectoral action are critical for pandemic response. It shows that civilian–military collaboration can be beneficial but contested, and that lack of fiscal space will undermine any response. It also demonstrates that pandemic preparedness cannot guarantee a successful pandemic response. Policy and research must pay more attention to improving decision-making processes when faced with pandemics involving novel pathogens, rapid spread, and substantial scientific uncertainty.
Levodopa use in Australia: an analysis of Pharmaceutical Benefits Scheme 10% data
Authors: Evans Andrew , Waterhouse Benjamin J 
Published: January 2024

BackgroundLevodopa remains the mainstay of treatment of Parkinson’s disease, however, over time motor fluctuations and levodopa-induced dyskinesia develop, requiring add-on therapies to control emerging symptoms. To date, however, there is no clear consensus in Australia, or elsewhere, at which dose of levodopa that add-on therapies should be considered.ObjectivesThe purpose of this study was to examine the treatment patterns of patients with Parkinson’s disease in Australia, with particular focus on levodopa doses at the time of first add-on.MethodsThis was a retrospective, observational, non-interventional study of patients with Parkinson’s disease within the Australian Department of Human Services Pharmaceutical Benefits Scheme (PBS) 10% sample. Data on all reimbursed prescriptions (both general and concession), prescriber type and item code were extracted for patients who were dispensed at least three PBS reimbursed prescriptions for levodopa in the previous 12 months prescription from 1 January 2007 to 31 December 2021.Results154 850 unique patients were included, of whom 42 330 (27%) commenced add-on therapy during the period. In the 12 months prior to add-on therapy, levodopa doses ranged from 100 mg/day to 1000 mg/day. The majority of patients were prescribed add-on therapy by a neurologist and approximately 40% of patients were prescribed levodopa doses of 600 mg/day or more prior to the first add-on therapy being initiated.ConclusionsA large proportion of patients in Australia are managed with levodopa monotherapy doses that are considered high and many of these patients may benefit from the addition of add-on therapy to their regimen.
Sudden unilateral audiovestibular loss due to acute labyrinthine haemorrhage can be missed on early MRI brain sequences: case report
Authors: Harrison Patrick , Blazak John , Richmond Joshua , Fraser-Kirk Kristy , Hoffmann Aliese , Collins Grant , Tsang Benjamin K-T 
Published: January 2024

BackgroundLabyrinthine haemorrhage is a rare vascular disorder often presenting with the triad of acute vertigo, sudden sensorineural hearing loss and tinnitus. There are minimal reports on imaging progression over the acute period.Index caseA woman in her mid-40s presented with acute vertigo, sudden left-sided hearing loss and tinnitus, consistent with acute unilateral audiovestibular loss. Left peripheral vestibular hypofunction was confirmed acutely on video head impulse testing, and pure tone audiometry showed a profound left sensorineural hearing loss. An MRI brain including diffusion-weighted imaging within 24 hours was normal. Delayed MRI brain and internal acoustic canal after 7 days demonstrated increased 3D fluid-attenuated inversion recovery and T1 signal throughout the left cochlea and semicircular canals, without contrast enhancement. This was consistent with labyrinthine haemorrhage. She received early oral prednisone followed by three doses of intratympanic dexamethasone. At 12 months follow-up the patient remained profoundly deaf, however, balance and vestibular symptoms improved with early vestibular physical rehabilitation.ConclusionWe report a case of acute labyrinthine haemorrhage missed on an early MRI brain sequence. This diagnosis should be considered in presentations of acute audiovestibular loss, and delayed MRI including internal auditory canal sequences may be important for diagnosis.
Randomised placebo-controlled trial of antenatal corticosteroids for planned birth in twins (STOPPIT-3): study protocol
Authors: Murray Sarah , Thompson Jessica , Townsend Rosie C , Deidda Manuela , Boyd Kathleen Anne , Norman Jane E , Norrie John , Boardman James P , Luyt Karen , Khalil Asma , Bick Debra , Reed Keith , Denton Jane , Fenwick Natasha , Keerie Catriona , Reynolds Rebecca , Stock Sarah Jane 
Published: January 2024

IntroductionThe aim of the STOPPIT-3 study is to determine the clinical and cost effectiveness of antenatal corticosteroids (ACS) prior to planned birth of twins in a multicentre placebo-controlled trial with internal pilot.Methods and analysisThis study will comprise a multicentre, double-blinded, randomised, placebo-controlled trial in at least 50 UK obstetric units. The target population is 1552 women with a twin pregnancy and a planned birth between 35 and 38+6 weeks’ gestation recruited from antenatal clinics. Women will be randomised to Dexamethasone Phosphate (24 mg) or saline administered via two intramuscular injections 24 hours apart, 24–120 hours prior to scheduled birth.OutcomesThe primary outcome is need for respiratory support within 72 hours of birth. Secondary and safety outcomes will be included. Cognitive and language development at age 2 years will be assessed in a subset of participants using the Parent report of Children’s Abilities-Revised questionnaire. We will also determine the cost effectiveness of the treatment with ACS compared with placebo.Ethics and disseminationSTOPPIT-3 has been funded and approved by the National Institute of Healthcare Research. It has been approved by the West Midlands Research Ethics Committee (22/WM/0018). The results will be disseminated via publication in peer-reviewed journals and conference presentation and will also be communicated to the public via links with charity partners and social media.Trial sponsorThe University of Edinburgh and Lothian Health Board ACCORD, The Queen’s Medical Research Institute, 47 Little France Crescent, Edinburgh, EH16 4TJ.Trial registration numberISRCTN59959611.