Early State Research Articles

Microwave Energy for Managing Hidradenitis Suppurativa

Title: Microwave energy for managing Hidradenitis Suppurativa Nicole Bielecki PA-C and Aaron Hoover MD Front Range Dermatology, Greeley, CO 80634 Introduction: Treating hidradenitis suppurativa (HS) is challenging due to its chronic, recurring nature and resistance to conventional therapies like antibiotics, anti-inflammatories, and surgery. Even with new biological treatments, there are challenges with injection site reactions. The associated pain, drainage, malodor, and scarring also significantly impact patients' mental well-being, underscoring the need for novel treatments. Four cases presented here suggest that microwave energy (ME) therapy can reduce pain, erythema and inflammation in HS lesions and even assist to resolve moderate and smaller lesions. Precise and brief surface-based applications of microwave energy to HS lesions from an FDA-cleared microwave device intended for the coagulation of soft tissue were used to generate localized hyperthermic conditions in HS and pre-HS lesions. The rapid heating of the tissues led to subsequent clearance of moderate and small HS lesions. This was accompanied with a reduction in inflammation, draining and pain associated with these HS cases. While the outcomes are promising, the exact mechanism of action needs further investigation. These cases strongly support further research and demonstrate that ME holds promise as a novel, non-invasive treatment modality for the management of HS. Procedure: A total of 25 HS lesions from five patients were treated with ME energy. Five repetitive 2-second pulses of 3 to 4W of ME operating at 8GHz were applied to the full area of these lesions. Three to four treatment sessions were given ranging from 7 to 14 days apart. Two patients received ME as a monotherapy, whereas the others began regimens of isotretinoin, adalimumab, or secukinumab at the same time as the first ME treatment. Results: These five patient cases demonstrate how ME treatments can lead to a rapid reduction in reported pain and erythema as well as clinical-assessed clearance of several early and moderate state HS lesions. Two of these patients only received ME as a monotherapy. However, based on clinical experience, even patients typically on medications will not typically show improvements within 7-14 days, suggesting that ME was having a rapid impact to manage pain and inflammation associated with HS lesions and in some cases assisting to resolve these lesions entirely. Conclusions: These cases report the first step in clinical feasibility and safety of microwave energy and shows promise as a novel modality for the management of HS. Although still in the early days of feasibility work, this non-invasive, fast, well-tolerated and simple treatment of microwave energy shows promise as a novel modality for the management of HS. Financial support: This work was supported by Emblation Ltd.

Abstract A007: Emergence of metastasis-initiating cells during lung adenocarcinoma progression

Abstract Metastatic lung adenocarcinoma (LUAD) is responsible for the most cancer-related deaths worldwide, and only a subset of cells is capable of metastasizing. Our lab has previously shown that these metastasis-initiating cells express the transmembrane glycoprotein called L1CAM and exhibit characteristics of a fetal lung developmental continuum, ranging from early progenitor states to more differentiated states. The exact signal by which progenitor cells with high metastatic potentials are generated remains unknown. Here, we demonstrate that L1CAM facilitates the signaling cascades necessary to initiate and maintain metastasis-initiating cells in LUAD. Mechanistically, L1CAM interacts with the planar cell polarity (PCP) complex and activates the transcription factor c-Jun. Activated c-Jun, as part of the AP-1 complex, binds to enhancer sites of progenitor genes and promotes the emergence of a metastasis-capable progenitor state. Pharmacological ablation of these metastatic progenitor cells by a cytotoxic L1CAM antibody effectively eliminated LUAD metastasis in vivo. Our findings identify a novel L1CAM-mediated mechanism that maintains a progenitor state essential for metastasis and can be targeted therapeutically. Citation Format: Jin Suk Park, Yasemin Kaygusuz, Carson Kenum, Mohamed I. Gatie, Lan He, Roshan Sharma, Ronan Chaligné, Abdul G. Khan, Ivo C. Lorenz, Paul J. Balderes, Tuomas Tammela, Anna-Katerina Hadjantonakis, Karuna Ganesh, Charles M. Rudin, Joan Massagué. Emergence of metastasis-initiating cells during lung adenocarcinoma progression [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A007.

Serum Calcium, Phosphate and Parathormone Levels in Thalassemic Children with High Ferritin Level

Background: In thalassemic children with high ferritin level due to iron accumulation in parathyroid gland may developed hypoparathyroidism leading to impaired homeostasis of calcium & phosphate. Parathyroid dysfunctions may not be evident clinically in early state. So the parathyroid status should be evaluated at regular interval. Objective: To find out the association of serum calcium, phosphate and parathormone levels in thalassemic children with high serum ferritin level. Materials and methods: The study was a cross-sectional study carried out in Department of Paeditarics, Dhaka Medical College Hospital, Dhaka, Bangladesh during January, 2019 to December, 2019. Total 70 thalassemia patients both male &female were included in this study between the age ranged from 5 to 12 years with serum ferritin level >1000 ng/ml irrespective of chelation therapy. All study subjects were fully evaluated clinically, thoroughly investigated and properly treated as per protocol. Results: Total 70 patients were included in our study. Male were 39 (55.7%) and female were 31(44.3%). S. Calcium level was decrease in 22(31.4%) patient, S PO4 level increased in 44 (62.9%) patients & parathyroid level was low in 16 (22.9%) patients. Conclusion: Based on result it could be concluded that thalassemic children age ranged from 5 years to 12 years, presenting with high ferritin level are likely to have low calcium, high phosphate & low parathormone level.

Identification of a Hidden, Highly Aggregation-Prone Intermediate of Full-Length TDP-43 That Triggers its Misfolding and Amyloid Aggregation.

In cells, TDP-43 is a crucial protein that can form harmful amyloid aggregates linked to fatal and incurable human neurodegenerative disorders. Normally, TDP-43 exists in a smaller soluble native state that prevents aggregation. However, aging and stress can destabilize this native state, leading to the formation of disease-causing amyloid aggregates via the formation of partially unfolded, high-energy intermediates with a greater tendency to aggregate. These intermediates are crucial in the early stages of amyloid formation and are challenging to study due to their low stability. Understanding the structure of these early aggregation-prone states of TDP-43 is essential for designing effective treatments for TDP-43 proteinopathies. Targeting these initial intermediates could be more effective than focusing on fully formed amyloid aggregates. By disrupting the aggregation process at this early stage, we may be able to prevent the progression of diseases related to TDP-43 aggregation. Hence, we decided to uncover the hidden, high-energy intermediates in equilibrium with the native states of TDP-43 by modulating the thermodynamic stability of the soluble native dimer (N form) and monomeric molten globular state (MG form) of full-length TDP-43. The thermodynamic modulation performed in the current study successfully revealed the highly aggregation-prone intermediate of full-length TDP-43, i.e., PUF. Moreover, we observed that along with high aggregation propensity, the aggregation kinetics and mechanisms of PUF differ from previously identified intermediates of full-length TDP-43 (the MG and I forms). The information regarding the initial aggregation-prone state of full-length TDP-43 could lead to therapies for amyloid diseases by halting early protein aggregation.

A narrative review of veterinary pharmacovigilance situations and prospects in East African countries

BackgroundVeterinary pharmaceuticals must be safe and effective for treating and preventing diseases in animal sectors. Strict adherence to regulations at every stage of production, storage, and administration is necessary as the global sector grows in order to lower the possibility of adverse veterinary drug events. Strong pharmacovigilance regulatory systems are critical in monitoring and managing safety concerns related with veterinary pharmaceuticals.ObjectiveThis review aimed to investigate the implementation of veterinary pharmacovigilance, collaborative initiatives, and reporting practices within the veterinary sector across East African countries.MethodsA thorough search was performed using online platforms such as Google Scholar, PubMed, the Web of Sciences, and regulatory Web sites. The search strategies relied heavily on selecting relevant published findings related to veterinary pharmacovigilance status, veterinary adverse drug event reporting practice, and collaborative efforts in veterinary pharmaceutical sectors within the East African landscape. This article search approach confirmed the inclusion state of veterinary pharmacovigilance and associated collaborative initiatives in the region.ResultsIn comparison to more developed regions, the review indicated that the veterinary pharmacovigilance system in East African countries was still in its early state. A strong legislative foundation and a large commitment from the veterinary profession are needed to establish a nationwide veterinary pharmacovigilance system. The review reveals a significant lack of consistency in the infrastructure of veterinary pharmacovigilance among the countries of East Africa. Tanzania, Kenya, and Ethiopia have some institutional processes for veterinary pharmaceutical safety, but they lack thorough documentation, which suggests that this systems still require improvement. The analysis emphasizes how inadequate the reporting systems are for adverse veterinary medication events in the majority of East African nations. Given the challenges East Africa faces, customized strategies are required to guarantee the safety and oversight of veterinary medications and improve veterinary pharmacovigilance. For systems to be more effective, veterinary pharmaceutical legal frameworks must be strengthened and stakeholder collaboration must be improved. Stakeholders include legislators, law enforcement, customs officials, regulatory organizations, scientists, pharmaceutical businesses, veterinarians, farmers, and the general public.ConclusionA multidisciplinary strategy is needed to solve current gaps in veterinary pharmacovigilance and improve overall veterinary drug safety in East Africa. It is feasible to create more reliable methods for tracking and guaranteeing the safety of veterinary medications by combining the efforts of numerous stakeholders, including as legislators, regulatory agencies, veterinary practitioners, and the general public.

Homeless Republicans: Haley Voters and the 2024 Presidential Primary in Coastal South Carolina

While former president Donald Trump cruised to victory in the 2024 Republican presidential primaries, he faced considerable opposition in the early states of Iowa, New Hampshire, and South Carolina. In these initial contests, former South Carolina governor Nikki Haley attempted to rally traditional conservatives in de

Multiomic analysis of familial adenomatous polyposis reveals molecular pathways associated with early tumorigenesis.

Familial adenomatous polyposis (FAP) is a genetic disease causing hundreds of premalignant polyps in affected persons and is an ideal model to study transitions of early precancer states to colorectal cancer (CRC). We performed deep multiomic profiling of 93 samples, including normal mucosa, benign polyps and dysplastic polyps, from six persons with FAP. Transcriptomic, proteomic, metabolomic and lipidomic analyses revealed a dynamic choreography of thousands of molecular and cellular events that occur during precancerous transitions toward cancer formation. These involve processes such as cell proliferation, immune response, metabolic alterations (including amino acids and lipids), hormones and extracellular matrix proteins. Interestingly, activation of the arachidonic acid pathway was found to occur early in hyperplasia; this pathway is targeted by aspirin and other nonsteroidal anti-inflammatory drugs, a preventative treatment under investigation in persons with FAP. Overall, our results reveal key genomic, cellular and molecular events during the earliest steps in CRC formation and potential mechanisms of pharmaceutical prophylaxis.

Evaluating Institutional Success: Regional Water Agreements with Tribal Nations

Abstract Since the early 1900s, U.S. national, tribal, and state institutions have been addressing conflicts over water entitlements of Native American tribes. The resulting regional water agreements encompass ensuring tribal access to water, defining entitlements for reliable future access, and providing environmental and financial benefits. Identifying approaches to evaluating institutional success in these regional water agreements is useful for effectively addressing current and future water disputes. This article discusses characteristics of eight water agreements that have occurred in the Southwestern United States, applying five economic criteria to evaluate success. The evaluation suggests that newer agreements perform better on the economic criteria, implying that learning about effective agreements is occurring over time. As threats to water quality and quantity intensify worldwide, understanding what contributes to successful regional water agreements becomes increasingly urgent.

Early state socialism and eugenics: Premarital medical certificates in Czechoslovakia, East Germany, and Poland in the aftermath of World War II

The article discusses the immediate post-war persistence and subsequent rejection of eugenics in East-Central European socialist states, exploring the case of premarital medical certificates. Building our analysis on published and archival sources, we show that immediately after the war, policies formulated at the governmental level were informed by eugenic ideas in medical expertise. Premarital medical certificates were aimed at combatting contagious diseases and thus securing a healthy population. Their legal status varied: in Poland, they were formally introduced; in the Soviet Occupied Zone and East Germany, Nazi law was abolished, but local officials still advocated for the introduction of certificates; and in Czechoslovakia, medical certificates were planned but not introduced. Despite these differences, after a short period between 1949 and 1950, the attempts to put a measure into practice ended in all three countries. We argue that the communist takeover and consolidation of power played a decisive role. After 1950, premarital medical certificates were not discussed again in Czechoslovakia or East Germany; in Poland, the debate re-emerged in 1959–60 during political liberalization.

Salivary Biomarker© In Biddable Hypertension Or Early Hyperglycaemic State Associated Ischemic Dilated Cardiomyopathy Related Myocardial Vessel Constriction

Salivary Biomarker© In Biddable Hypertension Or Early Hyperglycaemic State Associated Ischemic Dilated Cardiomyopathy Related Myocardial Vessel Constriction

Metabolic syndrome is linked to most cancers incidence.

Since many people die of either cancers or cardiovascular diseases worldwide, it is important to find the clinical pitfall that provokes cardiovascular diseases and cancer overall. Since metabolic syndrome (MetS) is largely linked to cardiovascular diseases, we have come to consider that MetS, even in its early state, may prime the occurrence of cancers overall. Indeed, the importance of MetS in causing pancreatic cancer has been proved using our large medical database. We analyzed Japanese healthcare and clinical data in 2005, who were followed up until 2020 and we examined the incidence of major cancers. At the enrollment, we examined the presence or absence of MetS judged by either Japanese criteria or NCEP/ATPIII. Of 2.7 million subjects without missing data, 102,930; 200,231; 237,420; 63,435; 76,172; and 2,422 subjects suffered lung, stomach, colon, liver and prostate cancer, respectively, and myelogenous leukemia during follow-up. MetS, defined by Japanese criteria, increased (p < 0.005 each) the incidence of cancer with a hazard ratio (HR) of 1.03-1.47 for lung, stomach, colon, liver, prostate cancers, and myelogenous leukemia. According to Japanese criteria, cancer incidence in the pre-stage MetS group was comparable to the MetS group. The results were almost identical when we defined MetS using NCEP ATP III. Taken together, we conclude that MetS is linked to majority of cancers.

State and law of the Chaghadai Ulus and early Timurid state as they were seen by foreign contemporaries

Research objectives: An analysis of official documents and notes of foreign contemporaries to clarify some aspects of political and legal life of the Chaghadai Ulus and early Timurid state in the 13th– beginning of 15th centuries. Materials and methods of research: The basic materials are official documents (diplomatic correspondence), travelers’ reports and notes, diaries and memoirs of travelers from different countries of Europe and Asia who visited the Chaghadai Ulus with different missions in the analyzed period as well as some official documents (diplomatic correspondence) between foreign and Mongol rulers. The methods of research are formal and historical legal research of historical documents, comparative legal analysis as well as historical approach in general. Scientific novelty: The state structure and legal relations in the Chaghadai Ulus and early Timurid state still have not been objects of special research. The author proposes to use works of foreign contemporaries as a tool to clarify some questions of political and legal life in the Chaghadaid state, finding similarities with other Chinggisid states and specific features of the region, its traditions, etc. Results of the research: The information of the analyzed historical documents, naturally, doesn’t allow one to reconstruct all aspects of political structure of Chaghadai Ulus and early Timurid state and their legal system in general, but adds valuable details to the information of other sources on history of this state – that is, historical chronicles (mainly of Timurid age) and legal monuments (khans’ yarliks and other official documents of the ulus). At that, we cannot ignore the fact that the Chaghadai Ulus before Timur was not considered as a full and authoritative member of international relations which is why contemporaries paid more attention to other Chinggisid states: The Golden Horde, Ilkhanate, Yuan Empire. However, the analysis of contemporaries’ notes could stimulate scholars to study some specific aspects of the Chaghadai Ulus’ history which were not researched before.

Research on the Spatial–Temporal Distribution and Morphological Characteristics of Ancient Settlements in the Sichuan Basin

An investigation into the spatial–temporal distribution characteristics of ancient settlements and their relationship with the environment in the Sichuan Basin can enhance our understanding of the evolution of the regional human–land relationship. Utilizing kernel density analysis by GIS, this paper obtained the spatial distribution characteristics of 1100 settlements in the Neolithic, Shang–Zhou, and Qin to Southern Dynasties periods. Subsequently, a GIS spatial analysis was performed to examine the distribution of these settlements in relation to the natural environment, focusing on four aspects: elevation, slope, aspect, and water buffer zone. The results demonstrate that: (1) The distribution of settlements from the Neolithic to the Southern Dynasties has expanded from the Chengdu Plain in the west and the Xiajiang Area in the east to the hilly areas in the center. (2) The settlements in the Neolithic and Shang–Zhou periods are predominantly located at elevations between 300–600 m, slopes between 0–3°, aspects between 135–225°, distances to rivers between 0–1000 m, and soil choice on Luvisols, Cambisols, Regosols, and Anthrosols. The influence of aspect on settlements during the Qin–Southern Dynasties period was notably reduced. The settlements mainly grow in the Chengdu Plain and the smaller plains along the river from the Neolithic to the Southern Dynasties in the Sichuan Basin. (3) The emergence and advancement of primitive agriculture, the construction of early water conservancy facilities, commercial and trade exchanges, and the establishment of regimes led to a shift from a double core to a more dispersed distribution in the Sichuan Basin from the Neolithic to the Southern Dynasties periods. The evolution of settlements reflects the adaption and competition of the residents to the natural environment in the Sichuan Basin. Furthermore, it provides insights into the complex social development in the region, ranging from the formation of early states to the establishment of the Qin and Han Empires.

Inhibition of PI3Kγ/δ signaling promotes an early memory state in CAR T cells and enhances their in vivo persistence and efficacy

Inhibition of PI3Kγ/δ signaling promotes an early memory state in CAR T cells and enhances their in vivo persistence and efficacy

The Assessment of Early Warning for Insurance Company Using Machine Learning Methods

Developing an early warning indicator is essential to strengthen the financial structure and take necessary precautions in a non-life insurance company. This paper aims to implement machine learning techniques on financial ratios to estimate the capital requirement ratio and investigate the solvency of an insurance company. For this purpose, the historical ratios of insurance companies in an emerging market are considered in accordance with the regulator’s solvency requirements. The ratios collected based on the performances of insurance companies in Türkiye are studied in two cases based on the number of features to be included in to four machine learning algorithms. “Full” data set with 69 and “Boruta” implemented data set with 33 ratios are employed to depict the efficiency of methods in predicting the early warning state of the company in terms of their capital requirement ratio predictions. Additionally, the assessment of these predictions to be utilized as an early warning indicator is performed. The findings illustrate that proposed early warning model predicts well the capital requirement ratio one year in advance. Moreover, among four ML methods, XGBoost achieves a prediction accuracy of 85% for estimating the state of the solvency in an insurance company compared to the other algorithms.

LineageVAE: reconstructing historical cell states and transcriptomes toward unobserved progenitors.

Single-cell RNA sequencing (scRNA-seq) enables comprehensive characterization of the cell state. However, its destructive nature prohibits measuring gene expression changes during dynamic processes such as embryogenesis or cell state divergence due to injury or disease. Although recent studies integrating scRNA-seq with lineage tracing have provided clonal insights between progenitor and mature cells, challenges remain. Because of their experimental nature, observations are sparse, and cells observed in the early state are not the exact progenitors of cells observed at later time points. To overcome these limitations, we developed LineageVAE, a novel computational methodology that utilizes deep learning based on the property that cells sharing barcodes have identical progenitors. LineageVAE is a deep generative model that transforms scRNA-seq observations with identical lineage barcodes into sequential trajectories toward a common progenitor in a latent cell state space. This method enables the reconstruction of unobservable cell state transitions, historical transcriptomes, and regulatory dynamics at a single-cell resolution. Applied to hematopoiesis and reprogrammed fibroblast datasets, LineageVAE demonstrated its ability to restore backward cell state transitions and infer progenitor heterogeneity and transcription factor activity along differentiation trajectories. The LineageVAE model was implemented in Python using the PyTorch deep learning library. The code is available on GitHub at https://github.com/LzrRacer/LineageVAE/.

The formation and development of concepts of access to court: a historical and legal context

The article is dedicated to the study of the elements of the formation and development of the judicial branch of power in specific historical periods in the context of outlining the principle of access to justice, particularly from the establishment of early states to modern times. In light of this, the article draws attention to the manifestations and signs of ensuring citizens' right of access to court from the perspective of studying the general historical progress of the judiciary. The analysis allowed the author to identify four main stages in the formation of the principle of access to justice in the world history of state and law: ancient, new, modern, and globalization. At the same time, the author emphasizes the different degrees of development and characteristic features of specific historical stages of the evolution of access to court and justice. It is established that already at the stage of formation and development of the ancient type of state and law (both Eastern and Western typologies), the formation of territoriality as one of the key components of the principle of access to justice began. Initially, judicial functions within the competence of the bodies and officials of the ancient states were combined with administrative and executive functions. The period of modern times is associated with the formulation of the political and legal idea of the separation of powers with the identification of an independent judiciary and the concept of due process, which is based on the modern notion of the ideal model of the organization and functioning of the judiciary in a democratic society. The rapid development and evolution of access to justice occurred in the modern stage, including the Florentine project of M. Cappelletti's "access to justice movement," which included a set of measures, aimed at ensuring real access to justice. The article concludes that the modern concept of access to court and justice involves its comprehension through empirical and pluralistic (multifaceted) approaches, taking into account not only obstacles and barriers to access to court and justice but also analytical data on the real state of affairs in the field of justice, considering various aspects of access to justice in the modern world

Association between Multi-Domain Lifestyle and Objective Cognitive Impairment in Elderly People with SCD and MCI in Chinese Communities.

Controlling the lifestyle associated with dementia risk can delay the process of cognitive decline. Subjective cognitive decline (SCD) and mild cognitive impairment (MCI) are early states in the development of dementia and are also the window period for early intervention in dementia. The purpose of this study was to explore the association between multi-domain lifestyle and objective cognitive impairment in elderly people with SCD and MCI in Chinese communities and to provide reference for effective implementation of precise health management measures to reduce the risk of dementia. A total of 265 middle-aged and elderly volunteers recruited from the community were divided into SCD group (107 cases), MCI group (80 cases), and healthy control (HC) group (78 cases). All participants received clinical interview, examination, and cognitive assessments. The total Dementia Risk Reduction Lifestyle Scale (DRRLS) scores in the HC, SCD, and MCI groups [110.00 (11.25) vs. 101.00 (10.00) vs. 79.50 (20.75)] exhibited statistically significant differences among them. The total score of the DRRLS showed a significant negative correlation with the Trail-Making Test (TMT), and significant positive correlations with both the Verbal Fluency Test (VFT) and Auditory Verbal Learning Test (AVLT) scores (p < 0.05). After adjusting for confounding factors, such as age and years of education, multiple linear regression analysis revealed several points. In the SCD group, brain-strengthening exercise and interpersonal relationship scores were negatively correlated with TMT scores (β = -11.257, -15.077; all p < 0.05), while health responsibility, smoking control behavior, and interpersonal relationship scores were positively correlated with AVLT scores (β = 0.485, 0.344, and 0.406; all p < 0.05). In the MCI Group, brain-strengthening exercise, brain-healthy diet, and interpersonal relationship were negatively correlated with TMT (β = -22.011, -16.206, -11.696; all p < 0.01), whereas health responsibility, mental activity, smoking control behavior, interpersonal relationship, and stress management were positively correlated with AVLT (β = 0.450, 0.435, 0.308, 0.256, 0.607; all p < 0.05). In Chinese communities, the unhealthy lifestyle of elderly individuals with SCD and MCI is significantly associated with cognitive function impairment. The greater their unhealthy lifestyle habits, the more pronounced the scope and severity of cognitive function impairment becomes. Furthermore, different dimensions of lifestyle have varying impacts on cognitive domains.

Development, evaluation and application of propidium monoazide (PMA) based methodologies on viable cell quantification of Pediococcus acidilactici in rice noodles products

Pediococcus acidilactici provides a special taste during dough fermentation, but its improper presence will accelerate the spoilage of rice noodles products. In this study, propidium monoazide (PMA) based methodologies on viable cells quantification of P. acidilactici were developed, evaluated and applied in rice noodles products. Prior to qPCR and RT-qPCR detection, PMA treatment was included to effectively remove the residual DNA from membrane-compromised (dead) cells. Firstly, considering the influence of food component and PMA on qPCR result determination, 4 standard curves corresponding Ct value to P. acidilactici cell number were established in 4 different rice noodle products, respectively. The 4 standard curves were subsequently applied for viable cell quantification of P. acidilactici. Secondly, mimicking the storage conditions of rice noodles products and growth states of P. acidilactici, planktonic and biofilm (early, mediate, mature) cells in 4 types of rice noodles products stored at −20 °C and 4 °C were taken into consideration to evaluate the efficiency of PMA-qPCR for viable cell quantification. Thirdly, stress response gene expression in P. acidilactici was monitored by PMA-RT-qPCR and compared with culturable and viable cell number quantification. The PMA-qPCR method with specific standard curves for 4 types of rice noodle products was applicable for viable cell quantification of P. acidilactici in planktonic and biofilm (early, mediate, and mature) states. The planktonic and biofilm cells of P. acidilactici maintained high and stable in both culturable and viable cell number in different growth states, rice noodles products, and low temperature conditions, but acquired significantly different gene expression levels. The findings provided viable cells quantification methods and an alarm for P. acidilactici contamination risk in instant rice noodles products.

Structural transition of GP64 triggered by a pH-sensitive multi-histidine switch

The fusion of viruses with cellular membranes is a critical step in the life cycle of enveloped viruses. This process is facilitated by viral fusion proteins, many of which are conformationally pH-sensitive. The specifics of how changes in pH initiate this fusion have remained largely elusive. This study presents the cryo-electron microscopy (cryo-EM) structures of a prototype class III fusion protein, GP64, in its prefusion and early intermediate states, revealing the structural intermediates accompanying the membrane fusion process. The structures identify the involvement of a pH-sensitive switch, comprising H23, H245, and H304, in sensing the low pH that triggers the initial step of membrane fusion. The pH sensing role of this switch is corroborated by assays of cell-cell syncytium formation and dual dye-labeling. The findings demonstrate that coordination between multiple histidine residues acts as a pH sensor and activator. The involvement of a multi-histidine switch in viral fusion is applicable to fusogens of human-infecting thogotoviruses and other viruses, which could lead to strategies for developing anti-viral therapies and vaccines.