bluuebunny/crossref_metadata_2025 huggingface dataset:
Mass Propagation of Nauclea diderrichii (de Willd. & Th. Dur) Merr. Seedlings through Tissue Culture Technique
Authors: Oyediran RI , Afolabi JO , Olomola DB , Akanni FO
Published: June 2021

Nauclea diderrichii is a tree species of economic importance. However, its plantation establishment is limited by inadequate seedling production. Hence, there is ample scope of tissue culture for its mass propagation. Its in vitro plantlets development as affected by media strengths indicated that 100 % seed germination was obtained in full MS basal medium while the least (3.35 %) was from quarter-strength at 8 Weeks after inoculation (WAI). The effects of BAP and NAA assessed on the growth of its sub-cultured plantlets showed that highest number of leaves (17) and adventitious shoots (3) were obtained from MS basal medium supplemented with 0.1 mg/l BAP only. Whereas, highest shoot length (3.61 cm) and average number of roots (5/plantlet) were obtained from the same medium without hormone(s) at 8 WAI. Further sub-culturing into MS with 0.05 mg/l NAA resulted into plantlets having optimum shoot and massive root growth ready for acclimatization in 6 WAI. The plantlets were successfully acclimatized using coconuthusk/ topsoil mixture with 90 % survival.

Plant Tissue Cult. & Biotech. 31(1): 51-60, 2021 (June)
The beginning phases of the early childhood caries (circular caries) - clinical evaluation of remineralization
Authors: Kokoceva-Ivanovska Olga
Published: June 2021

Early childhood caries (circular caries) can occur at an early age, with the eruption of deciduous teeth of the age of 1 to 1.5 years old. This is a multifactorial dental disease that has become a worldwide problem with high prevalence in recent decades. The purpose of the study was clinical evaluation of the remineralizing effect of the topical treatment, in patients with starting stages of circular caries.
This study was conducted at the Clinic for Pediatric and Preventive Dentistry, Skopje, and included 117 patients of both sexes, with the starting stages of circular caries. They were aged of 1.5 to 3.5 years old, with a fully formed primary tooth. It was selected 61 patients with initial lesion – white spot and 56 patients with superficial form. From both groups 30 patients were treated with topical fluoride preparation, for a period of six months, applied once a week. The remaining patients had regular check-ups and instructions, but they did not undergo fluoride treatment.
After the fluoride treatment, we followed the remineralizing effect, i.e. which initial lesions were reversed or where the intracrystal spaces were filled and the crystals were repaired after six months of topical application. In some patients who were able to satisfy other preventative factors, we were able to obtain a clinical picture of complete remineralization on initial lesion (restitution ad integrum) in vivo, approximately similar to a healthy tooth. How successful the remineralization will be depends on the predominant, protective, or pathogenic factors.

Keywords: circular caries, early childhood caries, remineralization, initial lesion, white spot
Concurrent genetic and standard screening test for hearing reduction
Authors: Davcheva Chakar Marina , Bozhinovski Gjorgji , Shukarova Stefanovska Emilija , Trajkov Dejan
Published: June 2021

Reduction of hearing is the most common sensory impairment among newborns with an incidence of 1-3 per 1000 births. Introduction of an Auditory Newborn screening program allows early identification of hearing impairment. Mainly, congenital hearing loss in early childhood is a result of genetic changes. Due to high frequency of GJB2 pathogenic variants, its molecular characterization among sensorineural hearing reduction cases is already conducted as a routine analysis in many countries. The aim of this study is to show our initial results in the effort to determine whether genetic screening along with the standard hearing screening in newborns is justified. Otoacoustic emission (OAE) method was conducted in 223 newborns at risk of hearing impairment. Among them, 7 did not pass the test in both ears while 9 exhibited one-sided hearing loss. In all 7 children with indication of profound bilateral deafness, the diagnosis was confirmed using auditory brainstem response. Genetic screening of GJB2 gene was performed in 6 of them. Genetic analysis of GJB2 revealed homozygous state of the most common pathogenic variant 35delG in 3 (50%) of the analyzed infants. In the remaining 3 no pathogenic variant was determined. The results indicate that performing auditory OAE together with genetic screening is justified. In newborns who have not passed the hearing screening test and have profound hearing loss, without other syndrome traits, screening for mutations of GJB2 gene should be conducted. Genetic screening enables establishment of early definite diagnosis for deafness and helps in conducting adequate therapy providing timely rehabilitation and social inclusion of deaf child.

Key words: hearing loss, genetic screening, auditory screening, GJB2 gene
Social media marketing of pharmacies across the Republic of North Macedonia
Authors: Pop-Nikolov Goran , Kapedanovska Nestorovska Aleksandra , Naumovska Zorica , Suturkova Ljubica , Sterjev Zoran
Published: June 2021

Internet marketing or digital marketing, is the fastest growing type of promotion for brands and companies to connect with potential customers. There are as many specializations within digital marketing as there are ways of interacting using digital media. This study evaluates and describes the usage of social media marketing as most common type of digital marketing used for self-promotion of the pharmacy sector in the Republic of North Macedonia. Social media marketing is raising brand awareness by engaging people in discussion online. The most popular platforms for social media marketing are Facebook and Instagram. For the purpose of the analysis, data from social network profiles Facebook and Instagram and from the websites of the 1000 pharmacies is used. This data includes: profile content, frequency of posts of health related articles and products, presence of relevant information and links to sites from which the content is shared, frequency of posting offers and sales at the pharmacy. The conducted research shows that the use of Internet as a digital marketing tool in Republic of North Macedonia is not sufficiently developed, especially when websites are used as a marketing tool for promotion of the pharmacy business and their services. The results show that Facebook as social media is used most frequently for posting health advices, promotion of new products and answering customer’s questions. Promotion of the professional pharmaceutical services is lower than the promotion of products. This leads to insufficient data for the implementation of the principles of evidence-based medicine in daily practice.

Keywords: Internet marketing, digital marketing, social media marketing, promotion, pharmacies, Facebook, Instagram, website
ULUSAL VE DIŞ BASINDA BAŞVEKİL İSMET PAŞA’NIN SOVYET RUSYA SEYAHATİ (26 NİSAN-9 MAYIS 1932)
Authors: ARSLAN Gürbüz
Published: June 2021

26 Nisan-9 Mayıs 1932 tarihleri arasında, Sovyet Rusya’da resmî temaslarda bulunan Başvekil İsmet Paşa, çeşitli kurumları ziyaret etmiş, yetkililerle görüşmeler yapmış, toplantı ve gezilere katılmıştır. Seyahat süresince, bir taraftan iki ülkenin rejiminin farklı olduğunu vurgulamış, diğer taraftan da bu durumun ikili ilişkilerin gelişmesine engel olmaması gerektiği üzerinde durmuştur. Buna karşılık Sovyet Rusya da benzer bir politika gütmüştür. Nitekim Sovyet Rusya’nın da İsmet Paşa’nın bu temasları esnasında, yakın ilişkiler içerisinde bulunduğu Türkiye’nin kendisinden uzaklaşmasını önlemek ve Batı’ya ihtiyacı olmadığını göstermek yönünde bir siyaset izlediği gözlemlenmiştir. Bu meyanda Moskova yönetimi, Türk yetkililere bütün kurumlarının kapılarını açmış, takip ettikleri ekonomik programı ve yöntemlerini göstermiştir. Yine bu bağlamda Sovyet Rusya’nın, Türkiye’nin sanayisini güçlendirmesi için ihtiyaç duyduğu krediyi ve dış politikada atacağı adımlara müdahale etmeyeceği teminatını verdiği görülmektedir.
Türkiye ile Sovyet Rusya arasındaki ilişkilerin işbirliği, dostluk ve güven çerçevesinde ilerlemesine vesile olan bu seyahat, yerli ve yabancı basın tarafından yakından takip edilmiştir. Bu çalışmada, İsmet Paşa’nın gerçekleştirdiği Sovyet Rusya ziyaretine yönelik yerli ve yabancı basındaki yansımaların, mukayese usulüyle bir değerlendirmesi yapılmıştır. Ayrıca çalışmada, TC Cumhurbaşkanlığı Devlet Arşivleri Başkanlığı Cumhuriyet Arşivi’ndeki Sofya ve Atina Büyükelçilerinin, seyahatle ilgili görev yaptıkları ülkelerin basın bültenlerinde yer alan haberlere yönelik göndermiş oldukları raporlardan da istifade edilmiştir.
İLK DEVİR TÜRK RESSAMLARINDAN BİR KESİT VE RESİM SANATINA KATKILARI
Authors: ÖZYİĞİT Halil
Published: June 2021

Osmanlı Devleti’nin son yüzyılı, kaybedilen savaşların etkisiyle hızlı ve farklı oluşumların yaşandığı bir döneme işaret etmektedir. Askerî ve eğitim gibi alanlarda bu yenilikler en güçlü şekilde karşılık bulur. Kültürel ve sanatsal aktivitelerde de bu değişim en üst seviyede hissedilir. Yaşanan bu değişim sürecini en yakından takip etmemizi sağlayan ise süreli yayınlardır. Osmanlı yazılı basının bu süreçte son derece etkili olduğunu, topluma yön vericiliğinin arttığını söyleyebiliriz. Bu bağlamda dönemin süreli yayınları gündemi tüm sıcaklığını okuyucularına yansıtan birincil kaynaklara dönüşmektedir. Osmanlı İmparatorluğu’nun bu döneminde Batı ile kurulan yakın temaslar, sadece siyasî ve ekonomik yapıyı değil, kültür ve sanat dünyasını da etkileyerek, bir değişim ivmesi yaratır. Bu değişimin ikinci aşaması ise Cumhuriyet’in kuruluşu ve gerçekleştirilen devrimlerle siyasi, sosyal, ekonomik ve kültürel yaşamın yeniden düzenlenmesidir. Bu makale, Batılı anlamda Türk resminin Osmanlı’dan Cumhuriyet’e uzanan süreçte anılan ilk devir Türk ressamlarından bir seçkiyi kapsamaktadır. Bu bağlamda Nami Dede, İbrahim Paşa, Hüsnü Yusuf Bey, Osman Nuri Paşa, Yusuf Ziya Paşa, Hüseyin Zekai Paşa ve Süleyman Seyyid Bey makalemizin ana karakterlerini oluşturmaktadır. Bu sanatçıların hem hayat hikâyeleri hem de sanatsal yönleri tek tek ele alınmıştır. Özellikle yaşam çizgilerine yakın tarihlerde yayınlanmış gazete, mecmua ve kitapların ışığında, dönemin ressamları ve üretimlerinin resim sanatımıza nasıl yön verdiğinin tespitine çalışılmıştır. Bu isimlerin Türk resim sanatına teknik, konu ve üslup olarak katkıları irdelenmeye çalışılacaktır. Türk resminin mihenk taşları arasında gösterebileceğimiz bu isimlerin büyük çabalarla açtığı yolda, Cumhuriyet’in feyz verdiği yeni ressamlar yürümeye başlayacaktır. Hiç şüphesiz Cumhuriyetin genç ve dinamik çocukları, seleflerinin kendilerine teslim ettiği sanat meşalesini çok daha ileriye taşıyacaklardır.
Characteristics of hematopoiesis in primary and recurrent ovarian cancer
Authors: Chulkova S. V. , Kozhonalieva A. M. , Stylidi I. S. , Poddubnaya I. V. , Palladina A. D. , Kolbatskaya O. P. , Zhordania K. I. , Kupryshina N. A. , Bokin I. I. , Artamonova E. V. , Egorova A. V. , Lepkova N. V. , Tupitsyn N. N.
Published: June 2021

Introduction. An important aspect of cancer treatment today is the concept of immuno-targeted therapy, which requires a deep understanding of the characteristics of immune reactions in the body of a cancer patient. Bone marrow is the central organ of immunopoiesis, therefore, along with the study of tumor characteristics, attention is paid to the bone marrow. The study of the cellular composition of the bone marrow in some types of cancer revealed a number of features of hematopoiesis, which requires further deeper analysis.Purpose. To study the parameters of hematopoiesis in patients with primary and recurrent ovarian cancer.Materials and methods. The paper presents data from 68 patients with a verified diagnosis of primary (n = 43) and recurrent (n = 25) ovarian cancer. The study was dominated by serous adenocarcinoma of high grade. Stage I was established in 13.2 % of cases, II – in 5.9 %, III – in 60.3 %, IV – in 20.6 %. The bone marrow was harvested by puncture of the posterior iliac spine (spina iliaca posterior superior). Parameter assessment and myelogram calculation were performed by two physicians, morphologists. The content of myelokaryocytes, indicators of granulocyte, erythroid lineage, the content of lymphocytes, monocytes were analyzed, and myelogram indices were assessed. In all patients microscopy of bone marrow samples excluded metastatic lesions. Statistical data processing was performed using the SPSS Statistics v. 21 package.Results. The analysis of bone marrow samples from patients with ovarian cancer revealed differences with the norm for both primary and recurrent ovarian cancer. Most punctates were normocellular, but average myelokaryocyte count in both groups was below normal. With recurrent ovarian cancer, a reduced content of promyelocytes and metamyelocytes was noted, whereas with primary cancer, all young forms of neutrophils was below normal. An increase in segmented neutrophils without a change in the percentage of cells of the granulocytic lineage was observed in primary ovarian cancer, and in one third of the samples of bone marrow an increased number lymphocytes and monocytes were noted. With recurrent ovarian cancer lymphocytes were increased in 2/3 of samples, monocyte – in 48 %. A change in the proportion of cells of the erythroid lineage was observed in both recurrent and primary cancers: depletion of the pool of basophilic normoblasts and polychromatophils with an unchanged number of erythrokaryocytes, an increase in oxyphilic forms were noted.Conclusion. The revealed changes in hematopoiesis in ovarian cancer reflect the aggressive course of high-grade tumors, which can be considered as a result of the systemic influence of the tumor, and the obtained data can serve as the basis for a detailed immunophenotypic study of bone marrow in ovarian cancer.
Assessing the effect of decopmression surgery performed due to malignant ischemic stroke on the quality of life
Authors: ŞİMŞEK Fatma , BİLGE Nuray , KOÇAK Mehmet Nuri , CEYLAN Mustafa , EREN Alper , YEVGİ Recep , KARAAVCI Nuh Çağrı
Published: June 2021

Aim: Decompressive craniectomy is one of the treatment methods that reduces mortality, when used in eligible patients for the treatment of malignant ischemic stroke (MIS). In this study, weaimed to assess the effect of age and the timing of surgery on the quality of life and on the prognosis of patients that underwent decompressive craniectomy.Material and Method: The files of 45 patients between the ages of 24-98, who were recommended decompressive craniectomy in connection with malignant ischemic stroke, were reviewed within the scope of the study. Result: 27 (60%) patients, who agreed to undergoing surgery, and 18 (40%) patients, who did not agree to undergoing surgery, were taken under review. A significant improvement (between 0 and 3) was observed in the modified Rankin Scale Scores of the two patients that underwent surgery. All the patients (n = 4) from within the group of patients that did not agree to a surgery, who lived for more than 6 months, were found to have a modified Rankin Scale Score of 5 and needed support to fulfill their daily life activities. Conclusion: It has been observed that performing a decompressive craniectomy after 72 hours following clinical deterioration and radiological shift did not have a significant effect on mortality and morbidity. Studies with larger patient groups are needed to be carried out in order to determine the applicability criteria of surgical treatment.
ANALISIS KUALITAS FISIK DAN DAYA TERIMA KONSUMEN TERHADAP PENGARUH PENGGUNAAN AMPAS KELAPA SEBAGAI PENGGANTI TEPUNG PANIR PADA NUGGET AYAM
Authors: Meitavani Meitavani , Mariani Mariani , Mahdiyah Mahdiyah
Published: June 2021

Penelitian ini bertujuan untuk mengetahui pengaruh penggunaan tepung panir ampas kelapa dengan tingkat kekasaran kasar, sedang, dan halus pada nugget ayam terhadap kualitas fisik dan daya terima konsumen. Metode yang digunakan adalah eksperimen dengan menggunakan uji organoleptik dan uji fisik. Hasil analisis uji daya terima konsumen pada 15 orang panelis agak terlatih menggunakan uji Friedman, didapatkan hasil terdapat pengaruh pada aspek warna, rasa, tekstur kulit, dan tekstur dalam, namun tidak terdapat pengaruh pada aspek aroma. Hasil uji lanjut Tuckey (α = 0,05) menunjukan bahwa terdapat perbedaan yang signifikan, namun terdapat kencenderungan nilai tertinggi, dimana perlakuan nugget ayam dengan tepung panir ampas kelapa halus merupakan perlakuan yang direkomendasikan pada aspek warna, rasa, tekstur kulit, dan tekstur dalam. Hasil uji fisik daya lekat menggunakan Anova menunjukan bahwa terdapat pengaruh tepung panir ampas kelapa pada daya lekat nugget ayam. Hasil uji lanjut dengan Duncan (α = 0,05) didapatkan bahwa nugget ayam dengan tepung panir ampas kelapa halus merupakan perlakuan terbaik pada aspek daya lekat. Uji fisik rendemen menggunakan Anova, didapatkan hasil terdapat pengaruh pada rendemen tepung panir ampas kelapa. Hasil uji Duncan menunjukan tepung panir ampas kelapa kasar merupakan perlakuan terbaik pada parameter rendemen. Produk nugget ayam dengan tepung panir ampas kelapa halus direkomendasikan sebagai perlakuan terbaik berdasarkan pertimbangan dari hasil kualitas fisik dan daya terima konsumen, serta tujuan utama untuk memanfaatkan dan meningkatkan nilai ekonomis ampas kelapa.
Daya Terima Konsumen Dan Sifat Fisik Roti Soft Roll Substitusi Tepung Hanjeli (C. Lacryma-jobi L.)
Authors: Irwan Irwan , Dahlia Mutiara , Ruslianti Rusilanti
Published: June 2021

ABSTRACT The aim of this research was to analyze the effect of job’s tear flour substitution in bread making on the consumer’s acceptance based on external (volume, crust’s color, form’s symmetry, bake’s evenness, crust characteristic) internal (grain, crumb’s color, aroma, flavor, texture) characteristics and physical characteristics (volume, growth power ratio). This research used experimental method. The sample of this research was soft roll with job’s tear substitution 10%, 20%, and 30% amount of percentages. Data collection technique used was acceptance test hedonic scale by 25 unskill panelists. The result of hedonic test indicated that soft roll with 10% job’s tear flour substitution was the most preferred on internal characteristics and soft roll with 20% job’s tear flour substitution was the most preferred on external characteristics. Friedman test with significant level α=0.05 showed that job’s tear flour substitution had effects on volume, crust’s color, crust characteristic, grain, aroma, flavor, and texture on soft roll. Tuckey’s test result showed that job’s tear flour substitution as much 10% was the most preferred and was recommended to be developed. Physical characteristics valuation showed that soft roll with 10% job’s tear flour substitution had bigger volume and growth power ratio than the other samples. ABSTRAKPenelitian ini dilakukan untuk mengetahui pengaruh substitusi tepung hanjeli dalam pembuatan roti soft roll terhadap daya terima konsumen pada aspek eksternal (volume, warna kulit roti, bentuk, keseragaman pemanggangan, karakter kulit roti) aspek internal (pori-pori roti, warna crumb, aroma, rasa, tekstur) dan sifat fisik roti (volume roti, rasio pengembangan). Penelitian ini menggunakan metode ekperimen. Sampel dalam penelitian ini adalah roti soft roll substitusi tepung hanjeli dengan persentase 10%, 20%, dan 30%. Teknik pengumpulan data yang digunakan adalah tes kesukaan oleh 25 panelis tidak terlatih. Hasil tes kesukaan menunjukan roti soft roll substitusi tepung hanjeli 10% merupakan produk yang paling disukai pada aspek internal dengan rentan nilai 4,00 hingga 4,24 dan roti soft roll substitusi tepung hanjeli 20% merupakan produk yang paling disukai pada hampir semua aspek eksternal dengan rentan nilai 4,24 hingga 4,40. Berdasarkan hasil uji Friedman dengan perbedaan signifikan α=0,05 menyatakan terdapat pengaruh substitusi tepung pada aspek volume, warna kulit roti, karakter kulit roti, pori-pori roti, aroma, rasa hanjeli, dan tekstur. Hasil uji Tuckey menunjukan bahwa roti soft roll substitusi tepung hanjeli 10% adalah yang disukai konsumen, sehingga direkomendasikan untuk dikembangkan. Penilaian fisik menunjukkan roti soft roll substitusi tepung hanjeli 10% memiliki volume dan rasio pengembangan terbesar dibandingkan sampel lainnya.
Healthcare Innovations and Ecosystems: COVID-19 and Beyond
Authors: Mukherjee Kanchan
Published: June 2021

This presentation gives an overview of the healthcare sector innovations and ecosystems in the Indian context. It introduces the concept of health, healthcare and healthcare innovation which is followed by the concept of innovation life cycle and ecosystem. The presentation shares the pre-COVID-19 and current situation in India in the context of healthcare innovations and ecosystems. It provides insights on the role of Health Technology Assessment (HTA) in health innovation ecosystems and the contribution of academic research institutes like the Tata Institute of Social Sciences (TISS) in HTA for healthcare innovation. The presentation concludes by acknowledging that the COVID-19 innovation sprint has now become a marathon, and hence healthcare innovations addressing systemic long term implications like equity and sustainability of healthcare systems become critical in the 'new normal' emerging from this pandemic. COVID-19 although a 'destructive creation' provides an opportunity for 'creative destruction' of voids for a sustainable inclusive future.
Sistem Pemantauan Parkir Liar Menggunakan Sensor Infrared Berbasis Android (Studi Kasus Politeknik Negeri Malang)
Authors: Wasian Nur Ahmad , Kusumawardani Mila , Perdana Ridho Hendra Yoga
Published: June 2021

Parkir liar yang berada di Politeknik Negeri Malang masih belum terpantau dengan baik. Petugas Pengamanan Dalam(Pamdal) yang bertugas untuk menertibkan warga Polinema memarkirkan kendaraan tidak dapat memantau kendaraan secara berkala, sehingga masih banyak yang melakukan parkir liar. Berdasarkan hal tersebut diperlukan sistem otomatis yang dapat digunakan Pamdal dalam memantau parkir liar di Polinema. Oleh karena itu dibuat “Sistem Informasi Pemantauan Parkir Liar Menggunakan Sensor Infrared Berbasis Android”. Sistem yang dibangun menggunakan beberapa komponen, diantaranya Arduino Nano, ESP32 Cam, Sensor Infrared, dan motor servo. Dengan dibuatnya perangkat dan penelitian ini diharapkan mampu membantu pendisiplinan warga polinema untuk tertib parkir melalui pemantauan dari sistem yang dibangun. Perangkat dibuat sebanyak tiga buah dan diletakkan di depan Gedung AH, di depan Gedung AI, dan di samping Gedung AO. hasil dari percobaan untuk notifikasi dari ketiga node memiliki delay yang berbeda pengiriman datanya dari node ke smartphone pada sisi user. Untuk Delay di node dua memiliki delay paling besar yakni sebesar 193.0708 ms. Sedangkan untuk node tiga memiliki performansi sangat baik dengan nilai rata-rata delay sebesar 107.0723 ms.
Lessons in Innate and Allergic Immunity From Dust Mite Feces and Tick Bites
Authors: Keshavarz Behnam , Erickson Loren D. , Platts-Mills Thomas A. E. , Wilson Jeffrey M.
Published: June 2021

Allergic diseases represent a major cause of morbidity in modern industrialized and developing countries. The origins and development of allergic immune responses have proven difficult to unravel and remain an important scientific objective. House dust mites (HDM) and ticks represent two important causes of allergic disease. Investigations into HDM fecal particles and tick bites have revealed insights which have and will continue to shape our understanding of allergic immunity. In the present review, focus is given to the role of innate immunity in shaping the respective responses to HDM and ticks. The HDM fecal particle represents a rich milieu of molecules that can be recognized by pathogen-recognition receptors of the innate immune system. Factors in tick saliva and/or tissue damage resultant from tick feeding are thought to activate innate immune signaling that promotes allergic pathways. Recent evidence indicates that innate sensing involves not only the direct recognition of allergenic agents/organisms, but also indirect sensing of epithelial barrier disruption. Although fecal particles from HDM and bites from ticks represent two distinct causes of sensitization, both involve a complex array of molecules that contribute to an innate response. Identification of specific molecules will inform our understanding of the mechanisms that contribute to allergic immunity, however the key may lie in the combination of molecules delivered to specific sites in the body.
Selective Anti-melanoma Effect of Phosphothioated Aptamer Encapsulated by Neutral Cytidinyl/Cationic Lipids
Authors: Wu Jing , Wang Shuhe , Li Xiang , Zhang Qi , Yang Jie , Ma Yuan , Guan Zhu , Yang Zhenjun
Published: June 2021

BC15-31 is a DNA aptamer that targets heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), which plays a crucial role in the process of pre-RNA maturation and is also essential for the rapid proliferation of tumor cells. In this research, we modified BC15-31 with a phosphorothioate (PS) backbone, LNA, and 2-O-MOE to enhance its stability and target affinity. In addition, a neutral cytidinyl lipid (DNCA) and a cationic lipid (CLD) were mixed to encapsulate modified aptamers with the aim of improving their cell permeability with low toxicity. Under the DNCA/CLD package, aptamers are mainly distributed in the nucleus. A modified sequence WW-24 showed an excellent selective anti-melanoma (A375 cells, ∼25 nM, 80%) activity, targeted to both hnRNP A1 and hnRNP A2/B1 found by the BLI experiment, and induced more early and late apoptosis in vitro, which also showed stronger antitumor effect and longer accumulation time in vivo. These results provide a new strategy for further clinical applications.
Tributyltin Oxide Exposure During in vitro Maturation Disrupts Oocyte Maturation and Subsequent Embryonic Developmental Competence in Pigs
Authors: Xiao Yue , Yuan Bao , Hu Weiyi , Qi Jiajia , Jiang Hao , Sun Boxing , Zhang Jiabao , Liang Shuang
Published: June 2021

Tributyltin oxide (TBTO), an organotin compound, has been demonstrated to have toxic effects on several cell types. Previous research has shown that TBTO impairs mouse denuded oocyte maturation. However, limited information is available on the effects of TBTO exposure on livestock reproductive systems, especially on porcine oocytes in the presence of dense cumulus cells. In the present research, we evaluated the effects of TBTO exposure on porcine oocyte maturation and the possible underlying mechanisms. Porcine cumulus-oocyte complexes were cultured in maturation medium with or without TBTO for 42 h. We found that TBTO exposure during oocyte maturation prevented polar body extrusion, inhibited cumulus expansion and impaired subsequent blastocyst formation after parthenogenetic activation. Further analysis revealed that TBTO exposure not only induced intracellular reactive oxygen species (ROS) accumulation but also caused a loss of mitochondrial membrane potential and reduced intracellular ATP generation. In addition, TBTO exposure impaired porcine oocyte quality by disrupting cellular iron homeostasis. Taken together, these results demonstrate that TBTO exposure impairs the porcine oocyte maturation process by inducing intracellular ROS accumulation, causing mitochondrial dysfunction, and disrupting cellular iron homeostasis, thus decreasing the quality and impairing the subsequent embryonic developmental competence of porcine oocytes.
In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
Authors: Afonin Sergii , Koniev Serhii , Préau Laetitia , Takamiya Masanari , Strizhak Alexander V. , Babii Oleg , Hrebonkin Andrii , Pivovarenko Vasyl G. , Dathe Margitta , le Noble Ferdinand , Rastegar Sepand , Strähle Uwe , Ulrich Anne S. , Komarov Igor V.
Published: June 2021

Labeling biomolecules with fluorescent labels is an established tool for structural, biochemical, and biophysical studies; however, it remains underused for small peptides. In this work, an amino acid bearing a 3-hydroxychromone fluorophore, 2-amino-3-(2-(furan-2-yl)-3-hydroxy-4-oxo-4H-chromen-6-yl)propanoic acid (FHC), was incorporated in a known hexameric antimicrobial peptide, cyclo[RRRWFW] (cWFW), in place of aromatic residues. Circular dichroism spectropolarimetry and antibacterial activity measurements demonstrated that the FHC residue perturbs the peptide structure depending on labeling position but does not modify the activity of cWFW significantly. FHC thus can be considered an adequate label for studies of the parent peptide. Several analytical and imaging techniques were used to establish the activity of the obtained labeled cWFW analogues toward animal cells and to study the behavior of the peptides in a multicellular organism. The 3-hydroxychromone fluorophore can undergo excited-state intramolecular proton transfer (ESIPT), resulting in double-band emission from its two tautomeric forms. This feature allowed us to get insights into conformational equilibria of the labeled peptides, localize the cWFW analogues in human cells (HeLa and HEK293) and zebrafish embryos, and assess the polarity of the local environment around the label by confocal fluorescence microscopy. We found that the labeled peptides efficiently penetrated cancerous cells and localized mainly in lipid-containing and/or other nonpolar subcellular compartments. In the zebrafish embryo, the peptides remained in the bloodstream upon injection into the cardinal vein, presumably adhering to lipoproteins and/or microvesicles. They did not diffuse into any tissue to a significant extent during the first 3 h after administration. This study demonstrated the utility of fluorescent labeling by double-emission labels to evaluate biologically active peptides as potential drug candidates in vivo.
Congenital Asplenia Interrupts Immune Homeostasis and Leads to Excessive Systemic Inflammation in Zebrafish
Authors: Xie Lang , Chen Zheyu , Guo Hui , Tao Yixi , Miao Xiaomin , Wu Ronghua , Li Yun
Published: June 2021

Splenectomy or congenital asplenia in humans increases susceptibility to infections. We have previously reported that congenital asplenia in zebrafish reduces resistance to Aeromonas hydrophila infection. However, the molecular mechanism of systemic immune response in congenitally asplenic individuals is largely unexplored. In this study, we found that pro-inflammatory cytokines were more highly induced in congenitally asplenic zebrafish than wild-type after pathogenic A. hydrophila infection and lipopolysaccharide exposure. In addition, a higher aggregation of apoptotic cells was observed in congenitally asplenic zebrafish than that in wild-type. Next, we examined the transcriptome profiles of whole kidneys from wild-type and congenitally asplenic zebrafish to investigate the effects of congenital asplenia on innate and adaptive immune responses induced by the inactivated A. hydrophila. Congenital asplenia inactivated the splenic anti-inflammatory reflex, disrupted immune homeostasis, and induced excessive inflammation as evidenced by the highly induced stress response–related biological processes, inflammatory and apoptosis-associated pathways, and pro-inflammatory cytokines/chemokines in congenitally asplenic zebrafish compared with wild-type after vaccination. In addition, complement component genes (c3a.1, c3a.6, c4, c6, and c9) and several important immune-related genes (tabp.1, tap1, hamp, prg4b, nfil3, defbl1, psmb9a, tfr1a, and sae1) were downregulated in congenitally asplenic zebrafish. Furthermore, congenital asplenia impaired adaptive immunity as demonstrated by downregulation of biological processes and signaling pathways involved in adaptive immune response after vaccination in congenitally asplenic zebrafish. The expression of MHCII/IgM was also significantly reduced in the congenitally asplenic zebrafish when compared with wild-type. Together, our study provides an in-depth understanding of spleen function in controlling immune homeostasis and may offer insight into the pathological response in splenectomized or congenitally asplenic patients after infections.
Effects of the Water-Sediment Regulation Scheme on the Expansion of Spartina alterniflora at the Yellow River Estuary, China
Authors: Fu Shuai , Zheng Shaoyan , Gao Weilun , Wang Andong , Ma Xu , Sun Limin , Sun Tao , Shao Dongdong
Published: June 2021

In recent decades, the invasion of saltmarsh plant Spartina alterniflora (S. alterniflora) over a large part of coastal wetlands in China, including the Yellow River Estuary (YRE) as a regional economic hub and global ecosystem services hotspot, has caused increasing concern because of its serious threats to native ecosystems. During the same period, local authorities have implemented a Water-Sediment Regulation Scheme (WSRS) in the Yellow River for flood mitigation and delta restoration purposes. The altered hydrological regime has resulted in unintended changes to estuarine ecosystem. However, the direct consequence of the WSRS on the expansion of S. alterniflora remains unclear. In this study, quantitative relationship between the inter- and intra-annual expansion patterns of S. alterniflora represented by relevant landscape metrics and indicators that quantify the concurrent variations of river and sediment discharges as the proxy of the WSRS impacts were analysed over the period of Year 2011–2018, and the analyses were performed on the YRE as a whole and on five different zones subdivided based on the invasion sequence. The results showed that there was no significant difference in the inter-annual area variation of S. alterniflora between the years with and without WSRS. Compared with the years without WSRS (2016–2017), the intra-annual (monthly) increment of the various landscape metrics [i.e., NP (number of patches), CA (class area), LPI (largest patch index) and AI (aggregation index] were found to be significantly higher in the initial stage of peak growing season (June-July) than in the mid- and late stages (July-September) in the years with WSRS (2011–2015, 2018) in the subregion located close to the south bank of YRE as the most prominent impact zone. In addition, F (mean flow), Ff (number of high flow pulses), Tf (Julian date of maximum flow) and D (duration of WSRS) were identified as the explanatory variables for the intra-annual vegetation landscape pattern changes, and their relative contributions to resultant changes were also assessed. Our results broaden the understanding of estuarine hydrological disturbance as a potential driver regulating the saltmarsh vegetation, and also have implications for S. alterniflora invasion control at estuaries under changing environment.
The Impacts of Two Revisions of the China’s Environmental Protection Fee on Firm Performance: Evidence From Stock Markets
Authors: Zheng Huan , He Yu
Published: June 2021

This comparative study aims to investigate how China’s environmental protection fee affected firm performance in two separate sample periods, namely 2001–2006 and 2012–2017, as this policy was revised twice in 2003 and 2014. With the Difference-in-Differences (DID) estimation, we find that the second revision of environmental protection fee had a negative impact on heavy polluting firms, while the influence of the first revision seemed to be insignificant. Moreover, the environmental protection fee had a limited impact on other firms, implying that such a policy had achieved its designed effect by precisely governing heavy polluters. Besides, our additional tests show that the environmental protection fee had stronger impacts on non-connected firms and non-SOEs than politically connected firms and SOEs. Our results are robust to various potential endogeneity issues.
Real-Time PCR Assays for Rapid Identification of Common Aphanomyces astaci Genotypes
Authors: Di Domenico Marco , Curini Valentina , Caprioli Riccardo , Giansante Carla , Mrugała Agata , Mojžišová Michaela , Cammà Cesare , Petrusek Adam
Published: June 2021

The oomyceteAphanomyces astaciis the etiologic agent of crayfish plague, a disease that has seriously impacted the populations of European native crayfish species. The introduction of non-indigenous crayfish of North American origin and their wide distribution across Europe have largely contributed to spread of crayfish plague in areas populated by indigenous crayfish. TrackingA. astacigenotypes may thus be a useful tool for investigating the natural history of crayfish plague in its European range, as well as the sources and introduction pathways of the pathogen. In this study, we describe the development of real-time PCR TaqMan assays aiming to distinguish the five genotype groups ofA. astaci(A–E) previously defined by their distinct RAPD patterns. The method was evaluated using DNA extracts from pureA. astacicultures representing the known genotype groups, and fromA. astaci-positive crayfish clinical samples collected mostly during crayfish plague outbreaks that recently occurred in Central Italy and Czechia. The assays do not cross-react with each other, and those targeting genotype groups A, B, D, and E seem sufficiently specific to genotype the pathogen from infected crayfish in the areas invaded byA. astaci(particularly Europe). The unusualA. astacigenotype “SSR-Up” documented from crayfish plague outbreaks in Czechia and chronically infectedPontastacus leptodactylusin the Danube is detected by the group B real-time PCR. The assay originally developed to detect group C (one not yet documented from crayfish plague outbreaks) showed cross-reactivity withAphanomyces fennicus; theA. astacigenotype “rust1” described in the United States fromFaxonius rusticusis detected by that assay as well. Analyses of additional markers (such as sequencing of the nuclear internal transcribed spacer or mitochondrial ribosomal subunits) may complement such cases when the real-time PCR-based genotyping is not conclusive. Despite some limitations, the method is a robust tool for fast genotyping ofA. astacigenotype groups common in Europe, both during crayfish plague outbreaks and in latent infections.
A Disrupted Historical Fire Regime in Central British Columbia
Authors: Brookes Wesley , Daniels Lori D. , Copes-Gerbitz Kelsey , Baron Jennifer N. , Carroll Allan L.
Published: June 2021

In the 2017 and 2018, 2.55 million hectares burned across British Columbia, Canada, including unanticipated large and high-severity fires in many dry forests. To transform forest and fire management to achieve resilience to future megafires requires improved understanding historical fire frequency, severity, and spatial patterns. Our dendroecological reconstructions of 35 plots in a 161-hectare study area in a dry Douglas-fir forest revealed historical fires that burned at a wide range of frequencies and severities at both the plot- and study-area scales. The 23 fires between 1619 and 1943 burned at intervals of 10–30 years, primarily at low- to moderate-severity that scarred trees but generated few cohorts. In contrast, current fire-free intervals of 70–180 years exceed historical maximum intervals. Of the six widespread fires from 1790 to 1905, the 1863 fire affected 86% of plots and was moderate in severity with patches of higher severity that generated cohorts at fine scales only. These results indicate the severity of fires varied at fine spatial scales, and offer little support for the common assertion that periodic, high-severity, stand-initiating events were a component of the mixed-severity fire regime in these forest types. Many studies consider fires in the late 1800s relatively severe because they generated new cohorts of trees, and thus, emphasize the importance of high-severity fires in a mixed-severity fire regime. In our study area, the most widespread and severe fire was not a stand-initiating fire. Rather, the post-1863 cohorts persisted due disruption of the fire regime in the twentieth century when land-use shifted from Indigenous fire stewardship and early European settler fires to fire exclusion and suppression. In absence of low- to moderate-severity fires, contemporary forests are dense with closed canopies that are vulnerable to high-severity fire. Future management should reduce forest densities and to restore stand- and landscape-level heterogeneity and increase forest resilience. The timing and size of repeat treatments such as thinning of subcanopy trees and prescribed burning, including Indigenous fire stewardship, can be guided by our refined understanding of the mixed-severity fire regime that was historically dominated by low- to moderate-severity fires in this dry forest ecosystem.
Identification of Clinically Relevant Subgroups of Chronic Lymphocytic Leukemia Through Discovery of Abnormal Molecular Pathways
Authors: Taus Petr , Pospisilova Sarka , Plevova Karla
Published: June 2021

Chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia in the Western world with a highly variable clinical course. Its striking genetic heterogeneity is not yet fully understood. Although the CLL genetic landscape has been well-described, patient stratification based on mutation profiles remains elusive mainly due to the heterogeneity of data. Here we attempted to decrease the heterogeneity of somatic mutation data by mapping mutated genes in the respective biological processes. From the sequencing data gathered by the International Cancer Genome Consortium for 506 CLL patients, we generated pathway mutation scores, applied ensemble clustering on them, and extracted abnormal molecular pathways with a machine learning approach. We identified four clusters differing in pathway mutational profiles and time to first treatment. Interestingly, common CLL drivers such as ATM or TP53 were associated with particular subtypes, while others like NOTCH1 or SF3B1 were not. This study provides an important step in understanding mutational patterns in CLL.
Identification of a Novel Immune-Related CpG Methylation Signature to Predict Prognosis in Stage II/III Colorectal Cancer
Authors: Chen Feng , Pei Lijuan , Liu Siyao , Lin Yan , Han Xinyin , Meng Erhong , Wang Xintong , Hong Shuai , Wang Dongliang , Liu Feide , Fei Yang , Wang Guangda
Published: June 2021

With the increasing incidence of colorectal cancer (CRC) and continued difficulty in treating it using immunotherapy, there is an urgent need to identify an effective immune-related biomarker associated with the survival and prognosis of patients with this disease. DNA methylation plays an essential role in maintaining cellular function, and changes in methylation patterns may contribute to the development of autoimmunity, aging, and cancer. In this study, we aimed to identify a novel immune-related methylated signature to aid in predicting the prognosis of patients with CRC. We investigated DNA methylation patterns in patients with stage II/III CRC using datasets from The cancer genome atlas (TCGA). Overall, 182 patients were randomly divided into training (n = 127) and test groups (n = 55). In the training group, five immune-related methylated CG sites (cg11621464, cg13565656, cg18976437, cg20505223, and cg20528583) were identified, and CG site-based risk scores were calculated using univariate Cox proportional hazards regression in patients with stage II/III CRC. Multivariate Cox regression analysis indicated that methylated signature was independent of other clinical parameters. The Kaplan–Meier analysis results showed that CG site-based risk scores could significantly help distinguish between high- and low-risk patients in both the training (P = 0.000296) and test groups (P = 0.022). The area under the receiver operating characteristic curve in the training and test groups were estimated to be 0.771 and 0.724, respectively, for prognosis prediction. Finally, stratified analysis results suggested the remarkable prognostic value of CG site-based risk scores in CRC subtypes. We identified five methylated CG sites that could be used as an efficient overall survival (OS)-related biomarker for stage II/III CRC patients.
Investigating Different DNA Methylation Patterns at the Resolution of Methylation Haplotypes
Authors: Peng Xiaoqing , Li Yiming , Kong Xiangyan , Zhu Xiaoshu , Ding Xiaojun
Published: June 2021

Different DNA methylation patterns presented on different tissues or cell types are considered as one of the main reasons accounting for the tissue-specific gene expressions. In recent years, many methods have been proposed to identify differentially methylated regions (DMRs) based on the mixture of methylation signals from homologous chromosomes. To investigate the possible influence of homologous chromosomes on methylation analysis, this paper proposed a method (MHap) to construct methylation haplotypes for homologous chromosomes in CpG dense regions. Through comparing the methylation consistency between homologous chromosomes in different cell types, it can be found that majority of paired methylation haplotypes derived from homologous chromosomes are consistent, while a lower methylation consistency was observed in the breast cancer sample. It also can be observed that the hypomethylation consistency of differentiated cells is higher than that of the corresponding undifferentiated stem cells. Furthermore, based on the methylation haplotypes constructed on homologous chromosomes, a method (MHap_DMR) is developed to identify DMRs between differentiated cells and the corresponding undifferentiated stem cells, or between the breast cancer sample and the normal breast sample. Through comparing the methylation haplotype modes of DMRs in two cell types, the DNA methylation changing directions of homologous chromosomes in cell differentiation and cancerization can be revealed. The code is available at: https://github.com/xqpeng/MHap_DMR.
Nutraceutical Approach to Preventing Coronavirus Disease 2019 and Related Complications
Authors: Derosa Giuseppe , Maffioli Pamela , D’Angelo Angela , Di Pierro Francesco
Published: June 2021

Introduction

Several months ago, Chinese authorities identified an atypical pneumonia in Wuhan city, province of Hubei (China) caused by a novel coronavirus (2019-nCoV or SARS-CoV-2). The WHO announced this new disease was to be known as “COVID-19”.

Evidence Acquisition

Several approaches are currently underway for the treatment of this disease, but a specific cure remains to be established.

Evidence Synthesis

This review will describe how the use of selected nutraceuticals could be helpful, in addition to pharmacological therapy, in preventing some COVID-19-related complications in infected patients.

Conclusions

Even if a specific and effective cure for COVID-19 still has some way to go, selected nutraceuticals could be helpful, in addition to pharmacological therapy, in preventing some COVID-19-related complications in infected patients.