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Blending coal before it enters the power plant boiler unit is necessary to adjust the coal categories according to the boiler unit specifications. The power plant must also comply with the regulations regarding coal-biomass co-firing through blending. Applying a Genetic Algorithm that only considers the composition and fitness based on the blend's quality leads to accumulation issues, decreasing coal quality. This research proposes redefining chromosomes, fitness functions, mutation rules, population determination, and output as the best chromosome used in the Genetic Algorithm. Testing uses various compositions of coal inputs from the barge, coal yard, and biomass to simulate different conditions. The test results demonstrate that the developed algorithm can provide all possible alternative blends between the coal in the barge and at the coal yard. Under specific conditions, operators can choose a blend composition that involves coal stored in the coal yard for an extended period.

More and more people are growing sweet potatoes in Hungary, but no one has yet tested how long they can be grown successfully on the same soil type. This means the quantitative and qualitative variables of the yield. From 2016 to 2022, I planted sweet potato slips in the same area and after four years, the yields decrease significantly. To reduce problems caused by soilborne pathogens, a three to five year rotation of crops is recommended. In Deszk, we set up the experiment on a 300 m2 area for all seven years. Based on clay loam soil. The area was fallowed for 5 years before 2016 and then planted with sweet potatoes for seven years. We always used the same field, so sweet potatoes were grown in monoculture. We also used the practices of growers in the area for soil preparation and the application of soil disinfectant. There was a significant difference in yields between seven years (p<0.05), with the highest yield (41.01 t/ha) in 2018, followed by 2016 (37.19 t/ha) and the lowest yield in 2022 (10.01 t/ha) (p<0.05). The total rainfall and average temperature of the 2017 growing season were not favourable, which was also reflected in the yield to a significant extent. The year 2019 was 30 t/ha, which can be considered as good results. From 2020 onwards, we have seen a continuous decrease in yields, which can be attributed to the effect of monoculture. The studies show that sweet potatoes can be grown successfully on clay loam soil for four years. In the following years the yield significantly decreased. Thus, it is advisable to set the field aside after four years. I recommend the use of ridges techology because using rides technology the harvesting is much easier, there will be fewer damaged tubers and the tubers have more room to grow in the ridges. Soil-dwelling pests need to be controlled, as soil-dwelling pests can cause serious problems in crop quality. It is advisable to change soil disinfectants every two years so that soil-dwelling pests do not become resistant to the active ingredient. [ABSTRACT FROM AUTHOR]

Despite the construction industry’s detrimental effects on environmental sustainability, the concept of green procurement (GP) is still relatively new in the developing world. The barriers hindering GP adoption need detailed investigation as studies in the context of developing countries are still limited and the underlying dimensions remain a known-unknown. This study appraised the critical barriers to GP adoption in the Malaysian construction industry. Twenty (20) barriers were identified from the detailed literature review and a field survey was conducted with 150 professionals. The ranking analysis results indicated the top five barriers are: high initial cost; lack of expertise and regulations; lack of government regulations; lack of top management commitment and lack of awareness among construction stakeholders. A factor analysis further identified five major underlying dimensions. The study showed that both government and organisational commitments are crucial in promoting GP adoption. This study adds to incremental knowledge in GP literature by analysing the critical barriers and the associated underlying dimensions to GP adoption in a developing country, which could help practitioners and policymakers in evaluating the organisation’s or industry’s readiness to adopt GP practices along with devising apt measures to mitigate the barriers to shift to sustainable and environmentally friendly construction.

Various accounting scandals involving the accounting profession are indicated because of the dominance of reason in accounting education. So, this research aims to understand and examine the perceptions of accounting educators about integrating Islamic values into accounting education. An interpretive perspective is used in this qualitative research. Data collection techniques through interviews. In-depth interviews were conducted with three accounting lecturers with open questions. Furthermore, the results of the interviews were transcribed and analyzed using the Miles and Huberman model, namely data reduction, data presentation, and conclusion. Islamic values are essential to be integrated into accounting education. Integrating Islamic values in accounting education includes praying together before learning, reading Quran verses, and practising "olah rasa" in class. The research results prove that Islamic values must be integrated into accounting education.

Xue Ting Tan, Nurliyana binti Mohd Shuhairi, Stephanie Jane Ginsapu, Surianti Binti Shukor, Fairuz Amran Bacteriology Unit, Infectious Diseases Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Selangor, MalaysiaCorrespondence: Xue Ting Tan, Bacteriology unit, Infectious Diseases Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Selangor, Malaysia, Tel +60 333628968, Email tanxt@moh.gov.myPurpose: This study was aimed to determine minimum inhibitory concentration (MIC) differences between yeast and mold forms of T. marneffei in Malaysia.Patients and Methods: Ninety-seven clinical strains of T. marneffei were received from various Malaysian hospitals from the year 2020 until 2022. Their identities were determined using microscopic, macroscopic and molecular methods. Next, the susceptibility of yeast and mold forms of each isolate against amphotericin B, itraconazole, voriconazole, posaconazole, ketoconazole, isavuconazole, terbinafine, caspofungin and micafungin were tested according to the broth microdilution according to the Clinical and Laboratory Standards Institute (CLSI) M38 and M27 guidelines. The geometric means of minimal inhibitory concentration (GM MIC), MIC50, and MIC90 were determined for each antifungal. Additionally, Wilcoxon signed-rank test was used to compare the significant difference of GM MICs for each antifungal, GM MIC, MIC50 and MIC90 for the combined nine antifungals against different growth forms of T. marneffei. The significance was set at p< 0.05.Results: Micafungin had the highest GM MIC, MIC50 and MIC90 for mold form of T. marneffei. For yeast form, amphotericin B achieved the highest GM MIC and MIC50 while micafungin achieved the highest MIC90. However, the GM MIC, MIC50 and MIC90 of terbinafine and azole antifungals on T. marneffei were similar to each other, namely between 0.03 and 0.60μg/mL. The difference of GM MIC of all tested antifungals except caspofungin and micafungin was insignificant. Overall, GM MIC, MIC50 and MIC90 of the combined nine antifungals against two growth forms were insignificant.Conclusion: The findings suggested either yeast or mold form can be used in the susceptibility testing of T. marneffei against amphotericin B, itraconazole, voriconazole, posaconazole, ketoconazole, isavuconazole and terbinafine.Keywords: Talaromyces marneffei, susceptibility, yeast, mold

Breast cancers are becoming harder to treat due to the acquisition of chemo-drug resistance. In this study, silver nanoparticles (AgNPs) were synthesized using reducing agent NaBH4, where resulting nanoparticles were characterized using UV-vis spectroscopy, FTIR spectroscopy, SEM, and DLS. Cytotoxicity of synthesized AgNPs was evaluated against MCF-7, MCF-7-CR, and MDA-MB-231 using MTT assays. NaBH4-reduced AgNPs were unstable as a colloidal system, with zeta potential noted to be around -21 mV and a polydispersity index of around 15%, making them highly prone to aggregation. However, AgNPs significantly reduced the cell viability of MCF-7 breast cancer cell lines, while slight toxicity was seen in multi-drug resistant breast cancer cells MCF-7-CR and MDA-MB-231 at 10 µM.

Cancer treatments have become less effective in recent years, with cancer cells mutating into more non-sensitive against a plethora of chemotherapeutics. While gene therapy can lead to cancer cell sensitization and effective treatment, gene-related drugs are prone to degradation and highly unstable. Nanoparticles can thus be used for the delivery of genes as well as other therapeutic compounds into cancer cells. Gold nanoparticles (AuNPs) have been used in cancer therapy especially for drug delivery due to their ease of synthesis and modification. AuNPs were also considered versatile nanoparticles as their characteristics could be easily customized and modulated for therapy, monitoring, or delivery. Another important characteristic that made AuNPs viable for cancer therapy is surface plasmonic resonance (SPR) which enables hyperthermia-based therapy and imaging. Therefore, utilizing AuNPs novel approaches in multi-modal therapies is achievable, where targeted gene and chemotherapy coupled with other modalities such as hyperthermia-based therapy could be performed. This article reviews several reported cases of AuNPs-based gene therapy as well as notable theranostic therapy that includes SPR characteristic utilization for therapy as well as cancer imaging.

This study aims to explain and construct the concept of cost beyond the reasoning in mainstream accounting. This construction is carried out by redefning the cost based on the “periphery” case. I reference to the Nusantara paradigm, the four principles (kaidah) in building Siraman Gong Kiai Pradah methodology have been fulflled. Therefore, the method used has been out of the ordinary research methods. First, it prepares a “periphery” case to be investigated for the costs. Second, it draws the position closer to God by doing penances so that they are given instructions in reducing and classifying the data. Third, doing the charity. Fourth, it includes the “value” of the “periphery” case in the new cost concept. Finally, this article proposes a change to the defnition of cost. Through the “periphery” case, the concept of cost transcends materialism. Cost is a number of expenses incurred, based on sincerity aimed at helping others in order to obtain the benefts both for the world and the hereafter by prioritizing the principle of mutual help.

This study aims to analyze the most important stakeholders in Indonesian companies. One of the main reporting principles in GRI 101 is stakeholder inclusiveness. Content analysis was used to identify stakeholders in 38 companies that participated in the 2017 Sustainability Reporting Awards. The first step taken is coding the stakeholders. The next step is to record the symbols and interpret the results according to the typology of stakeholder Mitchell et al (1997). The findings of the study show that shareholders and government are the most important stakeholders, regardless of industry differences in the sample. Abstrak Penelitian ini bertujuan untuk menganalisis stakeholder yang paling penting di perusahaan Indonesia. Salah satu prinsip pelaporan utama dalam GRI 101 adalah inklusivitas stakeholder. Analisis konten digunakan untuk mengidentifikasi stakeholder di 38 perusahaan yang mengikuti Sustainability Reporting Awards 2017. Langkah pertama yang dilakukan adalah melakukan coding terhadap stakeholder. Langkah selanjutnya adalah merekam simbol dan menafsirkan hasilnya sesuai kerangka typology stakeholder Mitchell et al (1997). Temuan penelitian menunjukkan bahwa pemegang saham dan pemerintah menjadi pemangku kepentingan yang paling penting, terlepas dari perbedaan industri dalam sampel.

A conformational epitope is a part of a protein-based vaccine. It is challenging to identify using an experiment. A computational model is developed to support identification. However, the imbalance class is one of the constraints to achieving optimal performance on the conformational epitope B cell prediction. In this paper, we compare several conformational epitope B cell prediction models from non-ensemble and ensemble approaches. A sampling method from Random undersampling, SMOTE, and cluster-based undersampling is combined with a decision tree or SVM to build a non-ensemble model. A random forest model and several variants of the bagging method is used to construct the ensemble model. A 10-fold cross-validation method is used to validate the model. The experiment results show that the combination of the cluster-based under-sampling and decision tree outperformed the other sampling method when combined with the non-ensemble and the ensemble method. This study provides a baseline to improve existing models for dealing with the class imbalance in the conformational epitope prediction.

PT. XYZ is a company that processes vaname shrimp into frozen shrimp products. Frozen shrimp is a product that serves for export, so that it must meet predetermined quality standards. Quality measurement is determined to monitor a process under controlled or uncontrolled conditions. The method of measuring is the six sigma method. The purpose of this research is an alarm that is responsive, alert, and alert, attentive, alert and responds to frozen products using the six sigma method at PT. XYZ. The frozen peeled shrimp production process stages start from receiving raw materials, cutting shrimp heads, quality sorting and size separation, peeling the skin and taking the intestines, soaking, freezing shrimp, packaging, metal detection, and product packaging. The results showed that 1,904 frozen peeled shrimp with 43 defective products had a sigma level of 3,22 with possible damage of 40.363,38 for one million products (DPMO) and the proportion of wrong causes of defects, namely 44,2%. The proposed improvements to be carried out are to provide direction and advice to employees to be careful and thorough, and the company offers a unique tool for cleavage of shrimp.

The COVID-19 pandemic has an impact on the economy of Padang City. To revive the economy, especially in the tourism sector, the government is trying to improve services to visitors so that the number of tourist visits increases again. As one of the efforts is to find out the opinion of tourists on the beach tourism area visited. This research aims to assist the Padang City government in knowing the positive or negative responses of tourists through the sentiment analysis process to the beach tourism they visit so that The Government of Padang City can determine the policies to be taken in connection with the reviews given by beach tourism visitors. By using reviews on Google Maps on the attractions of Air Manis Beach, Padang Beach, Pasir Jambak Beach, Nirwana Beach, and Pasir Putih Beach, clustering is carried out with the Naive Bayes classification algorithm. Based on the results of the analysis that has been done, 2 of the 5 beaches get negative reviews, namely Pasir Jambak Beach and Pasir Putih Beach which get negative values of 0.550 and 0.650.

Background A conformational B-cell epitope is one of the main components of vaccine design. It contains separate segments in its sequence, which are spatially close in the antigen chain. The availability of Ag-Ab complex data on the Protein Data Bank allows for the development predictive methods. Several epitope prediction models also have been developed, including learning-based methods. However, the performance of the model is still not optimum. The main problem in learning-based prediction models is class imbalance. Methods This study proposes CluSMOTE, which is a combination of a cluster-based undersampling method and Synthetic Minority Oversampling Technique. The approach is used to generate other sample data to ensure that the dataset of the conformational epitope is balanced. The Hierarchical DBSCAN algorithm is performed to identify the cluster in the majority class. Some of the randomly selected data is taken from each cluster, considering the oversampling degree, and combined with the minority class data. The balance data is utilized as the training dataset to develop a conformational epitope prediction. Furthermore, two binary classification methods, Support Vector Machine and Decision Tree, are separately used to develop model prediction and to evaluate the performance of CluSMOTE in predicting conformational B-cell epitope. The experiment is focused on determining the best parameter for optimal CluSMOTE. Two independent datasets are used to compare the proposed prediction model with state of the art methods. The first and the second datasets represent the general protein and the glycoprotein antigens respectively. Result The experimental result shows that CluSMOTE Decision Tree outperformed the Support Vector Machine in terms of AUC and Gmean as performance measurements. The mean AUC of CluSMOTE Decision Tree in the Kringelum and the SEPPA 3 test sets are 0.83 and 0.766, respectively. This shows that CluSMOTE Decision Tree is better than other methods in the general protein antigen, though comparable with SEPPA 3 in the glycoprotein antigen.

Abstract Background In Malaysia, leptospirosis is considered an endemic disease, with sporadic outbreaks following rainy or flood seasons. The objective of this study was to develop and validate a new knowledge, attitude, belief and practice (KABP) questionnaire on leptospirosis for use in urban and rural populations in Malaysia. Methods The questionnaire comprised development and validation stages. The development phase encompassed a literature review, expert panel review, focus-group testing, and evaluation. The validation phase consisted of exploratory and confirmatory parts to verify the psychometric properties of the questionnaire. A total of 214 and 759 participants were recruited from two Malaysian states, Kelantan and Selangor respectively, for the validation phase. The participants comprised urban and rural communities with a high reported incidence of leptospirosis. The knowledge section of the validation phase utilized item response theory (IRT) analysis. The attitude and belief sections utilized exploratory factor analysis (EFA) and confirmatory factor analysis (CFA). Results The development phase resulted in a questionnaire that included four main sections: knowledge, attitude, belief, and practice. In the exploratory phase, as shown by the IRT analysis of knowledge about leptospirosis, the difficulty and discrimination values of the items were acceptable, with the exception of two items. Based on the EFA, the psychometric properties of the attitude, belief, and practice sections were poor. Thus, these sections were revised, and no further factor analysis of the practice section was conducted. In the confirmatory stage, the difficulty and discrimination values of the items in the knowledge section remained within the acceptable range. The CFA of the attitude section resulted in a good-fitting two-factor model. The CFA of the belief section retained low number of items, although the analysis resulted in a good fit in the final three-factor model. Conclusions Based on the IRT analysis and factor analytic evidence, the knowledge and attitude sections of the KABP questionnaire on leptospirosis were psychometrically valid. However, the psychometric properties of the belief section were unsatisfactory, despite being revised after the initial validation study. Further development of this section is warranted in future studies.

Informal settlements are often not included in the development plan of cities. Urban planning has increasingly been less concerned about provision of basic infrastructure and services to vulnerable communities living and working in informal conditions. Increasing official apathy pose major challenges to achieving the globally accepted goal of inclusive urbanization as envisaged in Sustainable Development Goals / SDGs.. Goal 11 that aims at creating Safe, resilient, inclusive and sustainable cities will remain distant until we factor in the informal living and working conditions into our current urban planning narrative. GazdharBandh is a Notified Slum by Slum Redevelopment Authority (SRA) of MMRDA (Maharashtra Metropolitan Region Development Authority). The slum pocket is located in western suburb of Santacruz west, Mumbai. Gazdhar Bandh shows high density informal housing within the “no development zone” as per the development plan 2034, in estuary conditions that is vulnerable to recurrent floods. Slum upgradation faces multiple challenges with the complex interplay of local diversities, relocation and livelihood concerns. Using the case study of Gazdar Bandh slum, we examine how community responses to housing and other design strategies like public spaces, streetscapes, and makeshift spaces as immediate responses can go a long way to build long term resilience and factored into the broader city level planning narrative. The findings broadly examine the question: How can urban planning and design embrace uncertainty and concludes with an operational framework drawing on participatory planning and good urban place engaging multiple stakeholders.

This study is designed to solve the classroom problem dealing with the improvement of listening skill of the students of one private Islamic college that also study at pesantren in which they have lack of exposure in learning English. To cater a new way to overcome this issue, a classroom action research focusing on the implementation of interactive video viewing technique was employed. The findings exemplify that the students show better listening achievement after the implementation of second research cycle.

Pada penelitian ini diterapkan normalisasi dan pembobotan pada teknik pencampuran dengan metode Poisson sehingga dapat diperoleh citra hasil pencampuran. Dengan teknik ini, daerah batas antara dua citra yang dicampur dapat menyatu dengan sempurna tanpa harus melakukan penyuntingan gambar secara manual. Citra asal yang akan dicampur ke dalam citra target disegmentasi terlebih dahulu menggunakan metode ambang batas untuk membuat topeng. Metode Poisson diterapkan pada citra masukan dan target untuk mendapatkan nilai yang setelah dinormalisasi ke dalam [0 1] akan menjadi bobot dalam perhitungan nilai piksel baru pada daerah perbatasan antara citra asal dan target. Percobaan yang dilakukan pada sejumlah citra menunjukkan bahwa penggunaan bobot ternormalisasi hasil perhitungan Poisson memberikan efek keabuan yang makin meningkat mendekati batas luar citra asal. Hal ini menunjukkan bahwa prosentase nilai piksel pada batas terluar citra asal lebih besar dibandingkan dengan daerah batas bagian dalam. Perhitungan Kloning Mulus yang dilakukan dengan pembobotan tersebut menghasilkan gambar asal yang menyatu secara sempurna dengan citra target.

Geographical Information System (GIS) is one of the recent innovation to ease users in locating places. GIS is a system designed to capture, store, manipulate, analyze, manage, and present all types of spatial or geographical data. It can also be used by customers in locating business premises. Research related to application of GIS in locating SME business premises especially in Malacca as a tourist state is still very limited. This study used a survey method to produce more generalizable findings. Quantitative procedures for data collection and analysis were rigorously applied to ensure reliability and validity of the findings. A total of 160 usable responses was collected from 400 distributed questionnaires. This study found all variables significant and therefore, all hypotheses were accepted. In the future, researchers may extend this study to other areas to understand more about current usage patterns and reasons for its adoption. Studies which cover non-Malaysian respondents and different education levels can further document different perspectives of GIS adoption.

In this work, Al2O3 doped with Sr, Li and Ge was prepared by combustion synthesis techniques for thermoluminescent (TL) ionizing radiation dosimetry applications. Dopants sample concentration were varied from 0.1 mol% to 5 mol% in order to study the effect of the concentration on the TL response. The optimum concentration was found to be 0.3 mol% for Sr and Li and 3.0 mol% for Ge. Based from the comparison of this doped material, the highest sensitivity was found for Ge doped and linear response for doses studied range of 10-80 Gy. X-Ray diffraction (XRD) patterns of the samples were recorded to confirm the formation of the sample. The sharp peaks present in the XRD confirms the sample is crystalline. The TL response of these samples for Co -60 gamma radiation were evaluated. It was observed a TL glow peak around 175̊C for doped sample which is suitable for radiation dosimetry.

The conserved Caenorhabditis elegans protein kinases NEKL-2 and NEKL-3 regulate membrane trafficking and are required for larval molting. Through a forward genetic screen we identified a mutation in catp-1 as a suppressor of molting defects in synthetically lethal nekl-2; nekl-3 double mutants. catp-1 encodes a membrane-associated P4-type ATPase involved in Na+-K+ exchange. A previous study found that wild-type worms exposed to the nicotinic agonist dimethylphenylpiperazinium (DMPP) exhibited larval arrest and molting-associated defects, which were suppressed by inhibition of catp-1. By testing a spectrum catp-1 alleles, we found that resistance to DMPP toxicity and the suppression of nekl defects did not strongly correlate, suggesting key differences in the mechanism of catp-1-mediated suppression. Through whole genome sequencing of additional nekl-2; nekl-3 suppressor strains, we identified two additional coding-altering mutations in catp-1. However, neither mutation, when introduced into nekl-2; nekl-3 mutants using CRISPR, was sufficient to elicit robust suppression of molting defects, suggesting the involvement of other loci. Endogenously tagged CATP-1 was primarily expressed in epidermal cells within punctate structures located near the apical plasma membrane, consistent with a role in regulating cellular processes within the epidermis. Based on previous studies, we tested the hypothesis that catp-1 inhibition induces entry into the pre-dauer L2d stage, potentially accounting for the ability of catp-1 mutants to suppress nekl molting defects. However, we found no evidence that loss of catp-1 leads to entry into L2d. As such, loss of catp-1 may suppress nekl-associated and DMPP-induced defects by altering electrochemical gradients within membrane-bound compartments., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Genetics Society of America.)

Protein tyrosine phosphatases non-receptor type (PTPNs) have been studied extensively in the context of the adaptive immune system; however, their roles beyond immunoregulation are less well explored. Here we identify novel functions for the conserved C. elegans phosphatase PTPN-22, establishing its role in nematode molting, cell adhesion, and cytoskeletal regulation. Through a non-biased genetic screen, we found that loss of PTPN-22 phosphatase activity suppressed molting defects caused by loss-of-function mutations in the conserved NIMA-related kinases NEKL-2 (human NEK8/NEK9) and NEKL-3 (human NEK6/NEK7), which act at the interface of membrane trafficking and actin regulation. To better understand the functions of PTPN-22, we carried out proximity labeling studies to identify candidate interactors of PTPN-22 during development. Through this approach we identified the CDC42 guanine-nucleotide exchange factor DNBP-1 (human DNMBP) as an in vivo partner of PTPN-22. Consistent with this interaction, loss of DNBP-1 also suppressed nekl-associated molting defects. Genetic analysis, co-localization studies, and proximity labeling revealed roles for PTPN-22 in several epidermal adhesion complexes, including C. elegans hemidesmosomes, suggesting that PTPN-22 plays a broad role in maintaining the structural integrity of tissues. Localization and proximity labeling also implicated PTPN-22 in functions connected to nucleocytoplasmic transport and mRNA regulation, particularly within the germline, as nearly one-third of proteins identified by PTPN-22 proximity labeling are known P granule components. Collectively, these studies highlight the utility of combined genetic and proteomic approaches for identifying novel gene functions., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Binti et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Human NIMA-related kinases have primarily been studied for their roles in cell cycle progression (NEK1/2/6/7/9), checkpoint-DNA-damage control (NEK1/2/4/5/10/11), and ciliogenesis (NEK1/4/8). We previously showed that Caenorhabditis elegans NEKL-2 (NEK8/9 homolog) and NEKL-3 (NEK6/7 homolog) regulate apical clathrin-mediated endocytosis (CME) in the worm epidermis and are essential for molting. Here we show that NEKL-2 and NEKL-3 also have distinct roles in controlling endosome function and morphology. Specifically, loss of NEKL-2 led to enlarged early endosomes with long tubular extensions but showed minimal effects on other compartments. In contrast, NEKL-3 depletion caused pronounced defects in early, late, and recycling endosomes. Consistently, NEKL-2 was strongly localized to early endosomes, whereas NEKL-3 was localized to multiple endosomal compartments. Loss of NEKLs also led to variable defects in the recycling of two resident cargoes of the trans-Golgi network (TGN), MIG-14/Wntless and TGN-38/TGN38, which were missorted to lysosomes after NEKL depletion. In addition, defects were observed in the uptake of clathrin-dependent (SMA-6/Type I BMP receptor) and independent cargoes (DAF-4/Type II BMP receptor) from the basolateral surface of epidermal cells after NEKL-2 or NEKL-3 depletion. Complementary studies in human cell lines further showed that siRNA knockdown of the NEKL-3 orthologs NEK6 and NEK7 led to missorting of the mannose 6-phosphate receptor from endosomes. Moreover, in multiple human cell types, depletion of NEK6 or NEK7 disrupted both early and recycling endosomal compartments, including the presence of excess tubulation within recycling endosomes, a defect also observed after NEKL-3 depletion in worms. Thus, NIMA family kinases carry out multiple functions during endocytosis in both worms and humans, consistent with our previous observation that human NEKL-3 orthologs can rescue molting and trafficking defects in C. elegans nekl-3 mutants. Our findings suggest that trafficking defects could underlie some of the proposed roles for NEK kinases in human disease., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Joseph et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Molting is a widespread developmental process in which the external extracellular matrix (ECM), the cuticle, is remodeled to allow for organismal growth and environmental adaptation. Studies in the nematode Caenorhabditis elegans have identified a diverse set of molting-associated factors including signaling molecules, intracellular trafficking regulators, ECM components, and ECM-modifying enzymes such as matrix metalloproteases. C. elegans NEKL-2 and NEKL-3, two conserved members of the NEK family of protein kinases, are essential for molting and promote the endocytosis of environmental steroid-hormone precursors by the epidermis. Steroids in turn drive the cyclic induction of many genes required for molting. Here we report a role for the sole C. elegans ADAM-meltrin metalloprotease family member, ADM-2, as a mediator of molting. Loss of adm-2, including mutations that disrupt the metalloprotease domain, led to the strong suppression of molting defects in partial loss-of-function nekl mutants. ADM-2 is expressed in the epidermis, and its trafficking through the endo-lysosomal network was disrupted after NEKL depletion. We identified the epidermally expressed low-density lipoprotein receptor-related protein, LRP-1, as a candidate target of ADM-2 regulation. Whereas loss of ADM-2 activity led to the upregulation of apical epidermal LRP-1, ADM-2 overexpression caused a reduction in LRP-1 levels. Consistent with this, several mammalian ADAMs, including the meltrin ADAM12, have been shown to regulate mammalian LRP1 via proteolysis. In contrast to mammalian homologs, however, the regulation of LRP-1 by ADM-2 does not appear to involve the metalloprotease function of ADM-2, nor is proteolytic processing of LRP-1 strongly affected in adm-2 mutants. Our findings suggest a noncanonical role for an ADAM family member in the regulation of a lipoprotein-like receptor and lead us to propose that endocytic trafficking may be important for both the internalization of factors that promote molting as well as the removal of proteins that can inhibit the process., Competing Interests: The authors have declared that no competing interests exist.

Molting is a widespread feature in the development of many invertebrates, including nematodes and arthropods. In Caenorhabditis elegans, the highly conserved protein kinases NEKL-2/NEK8/9 and NEKL-3/NEK6/7 (NEKLs) promote molting through their involvement in the uptake and intracellular trafficking of epidermal cargos. We found that the relative requirements for NEKL-2 and NEKL-3 differed at different life-cycle stages and under different environmental conditions. Most notably, the transition from the second to the third larval stage (L2→L3 molt) required a higher level of NEKL function than during several other life stages or when animals had experienced starvation at the L1 stage. Specifically, larvae that entered the pre-dauer L2d stage could escape molting defects when transiting to the (non-dauer) L3 stage. Consistent with this, mutations that promote entry into L2d suppressed nekl-associated molting defects, whereas mutations that inhibit L2d entry reduced starvation-mediated suppression. We further showed that loss or reduction of NEKL functions led to defects in the transcription of cyclically expressed molting genes, many of which are under the control of systemic steroid hormone regulation. Moreover, the timing and severity of these transcriptional defects correlated closely with the strength of nekl alleles and with their stage of arrest. Interestingly, transit through L2d rescued nekl-associated expression defects in suppressed worms, providing an example of how life-cycle decisions can impact subsequent developmental events. Given that NEKLs are implicated in the uptake of sterols by the epidermis, we propose that loss of NEKLs leads to a physiological reduction in steroid-hormone signaling and consequent defects in the transcription of genes required for molting., (Copyright © 2022 Elsevier Inc. All rights reserved.)