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Abstract Background Precision-cut lung slices (PCLS) are ex vivo models with preserved lung cell populations and maintained tissue architecture. PCLS are, therefore, a powerful tool in respiratory research to study molecular mechanisms that closely reflect whole tissue biology. High-quality RNA and protein extraction from PCLS is, however, challenging as agarose significantly interferes with the yield and purity of extracted material. The present study aimed to optimize QIAzol-based isolation protocol for high-yield and quality RNA, miRNA, and protein extraction from PCLS. Materials and methods PCLS were prepared from 10 to 15-week-old Wistar rats and cultured for 7 days in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) supplemented with 0.1% FBS, penicillin, and streptomycin. LDH release to PCLS culture media was measured to determine cellular cytotoxicity. To select the optimal miRNA/RNA isolation protocol, we tested two different times (10 min, 2 h) and temperatures (room temperature, 4 °C, and −20 °C) of precipitation with isopropanol. Finally, we also assessed isolation with GHCL (guanidinium hydrochloride) extraction buffer. To select the optimal protein isolation protocol, we tested protein precipitation for 10 min at room temperatures (21 ± 1 °C) with 1.5 volumes of isopropanol and 3 volumes of acetone per 1 volume of phenol-ethanol supernatant. Additionally, we also tested protein precipitation for 3 h at −20 °C with 3, 5, and 7 acetone volumes per 1 volume of phenol-ethanol supernatant. We also validated protein precipitation with back extraction buffer instead of 100% ethanol. To measure the general efficiency of the optimized QIAZ-4 protocol, we used native rat lungs. PCLS for the ex vivo model of allergic inflammation were treated with IL-13 at a concentration of 80 ng/ml. Results Standard QIAzol isolation protocol provided RNA, miRNA, and protein with low yield and poor quality. We found that 2-h isopropanol precipitation at 4 °C with a high concentration of salts significantly increased the yield and quality of extracted RNA and miRNA and provided acceptable qPCR efficiency (between 90 and 110%). Surprisingly, 2-h isopropanol precipitation at −20 °C significantly increased qPCR efficiency above the acceptable range (average efficiency: 120.4%). As for protein extraction, we found that 3-h acetone precipitation at −20 °C provided the highest yield with linear protein detection on Westen Blot. Optimized QIAZ-4 provided significantly higher miRNA and RNA yield compared to standard QIAzol protocols. We also found a significantly increased expression of Eotaxin-1 in PCLS treated with IL-13 as compared to the untreated controls. Conclusions In our study, we described a simple QIAzol-based method for the simultaneous isolation of RNA, miRNA, and protein from PCLS.

IntroductionNormosmic isolated hypogonadotropic hypogonadism (nIHH) is a clinically and genetically heterogeneous disorder. Deleterious variants in over 50 genes have been implicated in the etiology of IHH, which also indicates a possible role of digenicity and oligogenicity. Both classes of genes controlling GnRH neuron migration/development and hypothalamic/pituitary signaling and development are strongly implicated in nIHH pathogenesis. The study aimed to investigate the genetic background of nIHH and further expand the genotype–phenotype correlation.MethodsA total of 67 patients with nIHH were enrolled in the study. NGS technology and a 38-gene panel were applied.ResultsCausative defects regarded as at least one pathogenic/likely pathogenic (P/LP) variant were found in 23 patients (34%). For another 30 individuals, variants of unknown significance (VUS) or benign (B) were evidenced (45%). The most frequently mutated genes presenting P/LP alterations were GNRHR (n = 5), TACR3 (n = 3), and CHD7, FGFR1, NSMF, BMP4, and NROB1 (n = 2 each). Monogenic variants with solid clinical significance (P/LP) were observed in 15% of subjects, whereas oligogenic defects were detected in 19% of patients. Regarding recurrence, 17 novel pathogenic variants affecting 10 genes were identified for 17 patients. The most recurrent pathogenic change was GNRHR:p.Arg139His, detected in four unrelated subjects. Another interesting observation is that P/LP defects were found more often in genes related to hypothalamic–pituitary pathways than those related to GnRH.ConclusionsThe growing importance of the neuroendocrine pathway and related genes is drawing increasing attention to nIHH. However, the underestimated potential of VUS variants in IHH etiology, particularly those presenting recurrence, should be further elucidated.

Abstract. This study is an attempt to build a theological view on the social activism of Pentecostals in Indonesia. Pentecostals in Indonesia are actually very familiar with social activities in the community. Otherwise, there would not be many Pentecostal Churches in various regions, which indicates the acceptance of the local community. However, such social activities are often understood separately from the understanding of the work of the Holy Spirit, or often understood to achieve the goal of proselytization. This study is conducted by re-reading the narratives of Joseph and Daniel in the Old Testament from the perspective of the Pneumatic work. The results of this study showed that the Spirit of God is the power that moves believers to work for the transformation of society. Abstrak. Kajian ini adalah sebagai upaya untuk membangun suatu pandangan teologis atas aktivisme sosial kaum Pentakostal di Indonesia. Kaum Pentakostal di Indonesia sebenarnya sudah sangat akrab dengan aktivitas sosial di tengah masyarakat. Jika tidak demikian, tidak mungkin banyak berdiri Gereja Pentakostal di berbagai daerah, yang mengindikasikan penerimaan masyarakat setempat. Namun, aktivitas sosial tersebut sering dipahami terpisah dengan pemahaman akan karya Roh Kudus, atau sering dipahami sebagai sarana mencapai tujuan proselitisasi. Kajian ini dikerjakan dengan membaca ulang narasi Yusuf dan Daniel di Perjanjian Lama dalam perspektif karya Pneumatologis. Hasil kajian ini menunjukkan bahwa Roh Tuhan adalah daya yang menggerakkan orang yang beriman untuk berkarya bagi transformasi masyarakat.

Abstract Normal and heavy water are solvents most commonly used to study the isotope effect. The isotope effect of a solvent significantly influences the behavior of a single molecule in a solution, especially when there are interactions between the solvent and the solute. The influence of the isotope effect becomes more significant in D2O/H2O since the hydrogen bond in H2O is slightly weaker than its counterpart (deuterium bond) in D2O. Herein, we characterize the isotope effect in a mixture of normal and heavy water on the solvation of a HOD molecule. We show that the HOD molecule affects the proximal solvent molecules, and these disturbances are much more significant in heavy water than in normal water. Moreover, in D2O, we observe the formation of low-density structures indicative of an ordering of the solvent around the HOD molecule. The qualitative differences between HOD interaction with D2O and H2O were consistently confirmed with Raman spectroscopy and NMR diffusometry.

Independence is very important for a child because independence fosters a sense of responsibility for children, independence bears joy, is closely related to children's emotional social intelligence, and independence contributes well to their future success. Judging from the position, closeness, and authority they have, fathers are considered the most appropriate in teaching this independence. Therefore, the active involvement of fathers is needed and expected. The research of this article is intended to explore the role that fathers can give in order to maximally teach independence to their children. By choosing a qualitative method with a description approach and support from a literature review, it is hoped that it can provide a maximum picture and be able to be accountable academically regarding independence in the biblical picture, the urgency of independence for children both now and in their future, as well as the contribution of fathers in teaching independence. It is concluded that fathers can contribute maximally in teaching independence to children when making it a habit for children, positioning themselves as companions in the learning process, being carried out with sensitivity and being a role model in terms of independence. Abstrak Kemandirian sangat penting bagi seorang anak karena kemandirian menumbuhkan rasa tanggung jawab anak, kemandirian berbuah kegembiraan, terkait erat dengan kecerdasan sosial emosional anak, serta kemandirian bersumbangsih baik pada kesuksesannya di masa depan nanti. Menilik posisi, kedekatan,dan otoritas yang dimilikinya, para ayah dirasa paling tepat dalam mengajarkan kemandirian ini. Maka dari itu keterlibatan aktif para ayah sangat dibutuhkan dan diharapkan. Penelitian artikel ini dimaksudkan ingin menelisik peran yang para ayah bisa berikan agar maksimal mengajarkan kemandirian pada anak. Dengan memilih metode kualitatif dengan pendekatan deskripsi dan sokongan dari kajian literatur diharapkan bisa memberikan gambaran yang maksimal serta mampu dipertanggung jawabkan secara akademik terkait kemandirian dalam gambaran Alkitab, urgensi kemandirian bagi anak baik itu di masa kini ataupun di masa depan mereka, juga kontribusi ayah dalam mengajarkan kemandirian. Disimpulkan bahwa para ayah dapat berkontribusi maksimal dalam mengajarkan kemandirian pada anak ketika menjadikannya sebagai pembiasaan bagi anak, memposisikan dirinya sebagai pendamping dalam proses belajar, dilakukan dengan kepekaan dan menjadi role model dalam hal kemandirian.

Abstract. This study was to explore the meaning of money in the Charismatic Pentecostal perspective. The method used was descriptive analysis through literature study. Through this study it was concluded that the Charismatic Pentecostals see the money is not merely needed to meet the daily life needs, it’s also as means of service to God. Service to God itself is not limited only in the form of ecclesiastical ministry, but also includes every work to promote a good life in this world. Abstrak. Penelitian ini berusaha mencermati pemaknaan uang dalam prespektif Pentakosta Karismatik. Metode yang digunakan adalah deskriptif analisis melalui kajian literatur. Berdasarkan kajian penelitian ini kaum Pentakosta Karismatik berpandangan bahwa uang selain dibutuhkan untuk mencukupi kebutuhan hidup sehari-hari, uang juga sebagai sarana pelayanan kepada Tuhan. Pelayanan kepada Tuhan itu sendiri tidak dibatasi hanya dalam bentuk pelayanan gerejawi, tetapi mencakup juga setiap karya untuk mewujudkan kehidupan yang baik di dunia ini.

Abstract: The Bible places the fear of God as the main foundation of believers' faith and provides examples of how the heroes of faith in the Bible carry out their daily lives in fear of God. This research explains how pantecostal theological reflection of the fear of God in people's lives is based on Psalm 25: 12-14. This article examines the Bible's view of the fear of God which is landed with real examples in the lives of several servants of God such as Abraham, Joseph, Job, David, Daniel and his three friends. This article also discusses the fear of God based on Psalm 25: 12-15 and Pentecostal theological reflection on the fear of God. Based on the results of the discussion, it was revealed that in the reflection of the Pentecostal theology of fearing God, an attitude that respects God, a choice taken by believers to live in God's blessings, the efforts made by believers to tie themselves to God's promises and finally believers to live in the way of the Lord. Abstrak: Alkitab meletakan takut akan Tuhan sebagai fondasi utama iman orang percaya dan memberikan contoh bagaimana para pahlawan iman di Alkitab menjalankan keseharian hidup mereka dalam takut akan Tuhan. Penelitian ini bermaksud menjelaskan bagaimana refleksi teologi pentakosta terkait sikap takut akan Tuhan dalam kehidupan orang percaya berdasarkan Mazmur 25: 12-14. Artikel ini mengulas tentang pandangan Alkitab tentang takut akan Tuhan yang didaratkan dengan contoh-contoh nyata dalam kehidupan beberapa hamba Tuhan seperti Abraham, Yusuf, Ayub, Daud, Daniel dan ketiga sahabatnya. Artikel ini juga membahas tentang karakteristik takut akan Tuhan berdasarkan Mazmur 25: 12-15 dan refleksi teologi Pentakosta tentang takut akan Tuhan. Berdasarkan hasil pembahasan disimpulkan bahwa dalam refleksi teologi Pentakosta takut akan Tuhan merupakan sikap yang menghormati Tuhan, sebuah pilihan yang diambil oleh orang percaya untuk hidup dalam berkat Tuhan, usaha yang dilakukan oleh orang percaya untuk mengikatkan diri pada janji Tuhan dan akhirnya menempatkan orang percaya untuk hidup dalam jalan-jalan Tuhan.

Children love languages is the language of expression and acceptance of the child. Through this love language, children express themselves and also measure the acceptance they get from the people around them. Every child has their own love language when communicating and parents are expected to understand their children's love language. The purpose of the research in this article is to understand the importance of using the language of love for children in Christian families. This research uses a descriptive method and literature review. The five languages of child love discussed in this article are physical touch, words of affirmation, quality time, acts of service, and gifts. Based on the results of this study, children's love language is important to be understood by parents because it is a medium for children's communication. By understanding the language of love for children, parents are expected to be able to prevent various crimes so that family harmony is maintained. Besides, understanding the children's love language will be able to increase the child's potential and become a family that other families can follow. Abstrak Bahasa cinta anak adalah bahasa ekspresi dan penerimaan anak. Melalui bahasa cinta ini anak-anak mengekspresikan dirinya sekaligus juga mengukur penerimaan yang dia dapat dari orang-orang disekitarnya. Setiap anak memiliki bahasa cintanya sendiri ketika berkomunikasi dan para orang tua diharapkan memahami bahasa cinta anak mereka. Adapun tujuan dari penelitian artikel ini adalah ingin memaknai arti penting penggunaan bahasa cinta anak dalam keluarga Kristen. Penelitian ini menggunakan metode deskripsi dan kajian literatur. Kelima bahasa cinta anak yang dibahas dalam artikel ini adalah sentuhan fisik, kata-kata peneguhan, waktu berkualitas, tindakan melayani dan hadiah. Berdasarkan hasil penelitian ini bahasa cinta anak penting untuk dipahami oleh para orang tua karena itu adalah media berkomunikasi anak. Dengan memahami bahasa cinta anak para orang tua diharapkan bisa mencegah berbagai kejahatan sehingga keharmonisan keluarga terjaga. Selain itu pemahaman bahasa cinta anak akan bisa meningkatkan potensi diri anak, dan menjadi keluarga yang bisa diteladani oleh keluarga lainnya.

Alkitab berasal dari Allah dan melibatkan tangan manusia dalam proses penulisannya. Alkitab umat Kristen sekarang ini terdiri dari dua bagian besar yaitu Perjanjian Lama dan Perjanjian Baru. Ketubim adalah bagian terakhir dari tiga bagian Alkitab Perjanjian Lama dalam kanon Ibrani. Artikel penelitian ini bermaksud memberikan penjelasan mengenai tujuan nubuat bagi umat pilihan Tuhan dalam Ketubim secara sistematik sehingga memudahkan untuk dipahami oleh para pembaca artikel ini. Metodologi yang digunakan oleh peneliti dalam artikel ini adalah metodologi kualitatif dengan pendekatan analisis teks dan kajian literatur. Artikel ini membahas secara mendalam mengenai nubuat Raja Daud tentang tuan, nubuat tujuh puluh kali tujuh masa Daniel, dan nubuat Mikha dalam peristiwa Raja Ahab memerangi Ramot-Gilead sebagai contoh-contoh nubuat dalam Ketubim disertai analisis berdasarkan bahasa asli serta didukung dengan literatur dari buku-buku dan jurnal ilmiah. Adapun berdasarkan hasil penelitian tujuan nubuat dalam Ketubim adalah memberikan pemahaman bahwa Yesus adalah Tuhan, menjadi suara kebenaran pada zamannya, menyatakan apa yang menjadi kehendak Tuhan bagi umat pilihan-Nya, dan penjelasan tentang Akhir Zaman.

Most neuroendocrine neoplasms (NEN) are characterized by the presence of somatostatin receptors (SSTR) which we use in location diagnostics and treatment. The aim of this study was to evaluate the expression of somatostatin receptors by immunohistochemistry in tissue obtained after surgery of the primary focus in the small intestine. The group of patients consisted of 41 people, in 18 cases the primary tumor was in the jejunum and in 23 in the ileum. The immunohistochemical method was used to visualize the receptors, using polyclonal antibodies in a two-stage peroxidase method. In patients with NEN of the small intestine, the SSTR2a and SSTR5 receptors are most commonly expressed, followed by SSTR2b and 3. In statistical analysis, it was shown that the expression of somatostatin receptors was not dependent on the primary site of the tumor (p > 0.05). The dependence of SSTR expression on histological maturity is evident. SSTR1, SSTR2b, SSTR3 and SSTR5 are more common in tumors with grading G1 (p

Abstract. Church growth can be achieved through church’s member migration, biological growth, and also evangelistic missions. For certain churches, evangelistic missions are not only one way to increase the number of congregations, but are seen as part of the fulfillment of the Great Commission. The purpose of this study was to describe the role of missionary evangelism in church growth. The research method used is a qualitative method with a descriptive approach. Through this research, it was obtained that the mission of evangelism is an effective means of increasing church growth. The mission of evangelism could be carried out with various strategies that are appropriate to the characteristics of the community around the church to be able to deliver the gospel effectively. Abstract. Pertumbuhan gereja dapat dicapai melalui perpindahan jemaat, pertambahan secara biologis, maupun misi penginjilan. Bagi gereja tertentu misi penginjilan bukan hanya sebagai salah satu cara untuk menambah jumlah jemaat, namun dipandang sebagai bagian penemuhan Amanat Agung. Tujuan penelitian ini untuk memberikan gambaran peranan misi penginjilan terhadap pertumbuhan gereja. Metode penelitian yang digunakan adalah metode kualitatif dengan pendekatan deskriptif. Melalui penelitian ini diperoleh gambaran bahawa misi penginjilan adalah sarana yang efektif untuk meningkatkan pertumbuhan gereja. Misi penginjilan tersebut dapat dilakukan dengan berbagai strategi yang sesuai dengan karakteristik masyarakat di sekitar gereja untuk dapat menyeberangkan Injil secara efektif.

Abstract. The purpose of this study was to evaluate the phenomenon of prophecy in Charismatic churches. The method used in writing this atricle was combining the analysis of the prophetic texts of Paul's writings and the study of phenomenology. Through this research it was revealed that the practice of prophecy is widely practiced among Charismatic churches in the city of Semarang. The results showed that the pastors in the Charismatic churches need to have a correct understanding of the practice of prophecy, especially understanding Paul's teaching about prophecy, in order to avoid the adverse effects for the church caused by the practice of prophecy. Abstrak. Tujuan penelitian ini adalah untuk mengevaluasi fenomena nubuatan yang marak di gereja Karismatik. Ada pun metode yang digunakan dalam penulisan atrikel ini adalah memadukan analisis teks-teks nubuatan tulisan Paulus dan studi fenomenologi. Melalui penelitian ini terungkap bahwa praktik bernubuat banyak dipraktikan di kalangan gereja beraliran Karismatik di kota Semarang. Hasil penelitian menunjukkan bahwa pendeta-pendeta di kalangan gereja Karismatik perlu memiliki pemahaman yang benar tentang praktik bernubuat, khususnya memahami dengan benar pengajaran Paulus tentang nubuatan, supaya terhindar dari dampak yang merugikan gereja yang disebabkan oleh praktek nubuatan tersebut.

Penelitian ini bermaksud mengkaji strategi orang tua dalam menumbuh kembangkan disiplin anak di keluarga Kristen. Alkitab meletakan tanggung jawab utama untuk mendidik anak ada pada orang tua. Anak-anak memerlukan disiplin untuk membentuk karakter mereka. Selain itu disiplin juga penting bagi anak dalam kaitan dengan kehidupan sosial dan disiplin merupakan fondasi yang sangat dibutuhkan anak untuk meraih masa depan mereka. Metode yang digunakan adalah deksriptif dan kajian literatur. Berdasarkan hasil pembahasan artikel ini tersimpulkan bahwa para orang tua berperan dalam membangun disiplin anak mereka yaitu melalui membangun komunikasi dengan menggunakan bahasa cinta dari anak mereka, dengan menjadikan dirinya teladan yang bisa dicontoh oleh anaknya, memberikan ruang untuk anak bertumbuh serta mengeksplore dirinya, dan melalui tindakan tegas dalam kasih.

Artikel ini bermaksud memberi pemaknaan terhadap kemarahan Allah dalam sudut pandang teologi Pentakosta. Metodologi yang digunakan yaitu analisis teks dan kajian literatur. Ada empat contoh kemarahan Allah yang diteliti yaitu kemarahan Allah kepada Adam dan Hawa, kemarahan Allah kepada bangsa Israel, kemarahan Tuhan Yesus kepada para murid, dan kemarahan Tuhan Yesus kepada para pedagang di Bait Allah. Hasil penelitian menunjukkan bahwa Allah tidak pernah marah tanpa alasan yang jelas dan ada dampak dari setiap kemarahan Allah. Penyebab kemarahan Allah adalah ketidaktaatan, menggantikan Allah dengan ilah lain, ketidakadilan, dan memperjualbelikan pelayanan. Adapun dampak kemarahan Allah adalah keterpisahan dengan Allah, kehilangan komunitas ilahi, mendatangkan penghukuman Allah, dan kematian.

Diabetes insipidus is a disorder resulting from insufficient action of vasopressin (ADH) characterized by excretion of highly diluted urine in large amounts. Idiopathic diabetes insipidus is associated with the presence of both autoantibodies against ADH-secreting neurons and pituitary autoantibodies. The aim of the present study was to evaluate the occurrence of autoantibodies against the pituitary microsomal fraction. The study included 33 sera of diabetes insipidus patients and 10 control sera obtained from 10 healthy persons. In all patients the secretion of pituitary hormones and thyroid autoantibodies was assessed. Human pituitaries were obtained during autopsy and homogenized in 0.01 mol/l pH 7.4 phosphate buffer. In addition, for the autoantibody evaluation, the electrophoretic method of separation in polyacrylamide gel and western blot were employed. Among the 33 subjects, in 23 patients the presence of autoantibodies against the pituitary was shown. Sera of 15 patients reacted with the pituitary microsomal fraction protein of 55 kDa. In other cases, 10 sera reacted with the pituitary antigen of 67 kDa. In addition, 5 sera reacted with the 60 kDa antigen, 5 sera with 52 kDa protein, 3 sera with 105 kDa protein, 3 sera with the 97 kDa antigen and 2 sera with pituitary antigen of 92 kDa weight. In our study, based on the immunoblotting method, we observed that pituitary autoantibodies against 55, 60 and 67 kDa antigens occurred frequently.

Introduction Problems related to eating disorders such as anorexia, bulimia and obesity turns out to be very common in the LGBT group. The paper presents a review of research in this field showing the relationship between the influence of the environment, homophobia and lack of tolerance to the problem presented in the topic and a scale of this problem occurring in the presented minority group. Aim The aim of the work is to draw attention to the scale of the problem and the specificity of the LGBT group in the field of eating disorders. Presenting factors which are a predisposition to such disorders, may be useful to medical staff, psychologists, educators and therapists dealing with the problem of eating disorders. Another aim is to create a scientific publication on the topic of sexual minorities, which is still lacking in comparison with foreign magazines in our country. Brief description of the state of knowledge LGBT group turns out to be a group with an increased risk of eating disorders. Gays are a group exposed to anorexia and bulimia. However, the group of lesbians has been defined with an increased risk of overweight and obesity. According to a review of contemporary research in this area, the problems related to nutrition are influenced by homophobia, lack of social acceptance, heterosexual and episodes of sexual abuse in childhood. Conclusions There is a huge need of knowledge for specialists dealing with eating disorders about the specificity of the LGBT group. Their greater competences can positively influence the prevalence of eating disorders in this group and positively influence the course of treatment of such people.

Hemagglutinin (HA), the class I influenza A virus protein is responsible for the attachment of virus particles to the cell by binding to glycan receptors, subsequent virion internalization, and cell entry. Consequently, the importance of HA makes it a primary target for the development of anti-influenza drugs. The natural oligoribonucleotides (ORNs) as well as their derivatives functionalized with D-mannitol (ORNs-D-M) possess anti-influenza properties in vitro and in vivo due to interaction with HA receptor sites. This activity suppresses the viral infection in host cells. In the present work, the complexes of ORNs and ORNs-D-M with HA protein were studied by agglutination assay, fluorescence spectroscopy, as well as molecular docking simulations. Acquired experimental data exhibited a decrease in HA titer by 32 times after incubation with the ORNs-D-M for 0.5–24 h. Quenching fluorescence intensity of the HA suggests that titration by ORNs and ORNs-D-M probably leads to changes in the HA structure. Detailed structural data were obtained with the molecular docking simulations performed for ORNs and ORNs-D-M ligands containing three and six oligoribonucleotides. The results reveal that a majority of the ORNs and ORNs-D-M bind in a non-specific way to the receptor-binding domain of the HA protein. The ligand’s affinity to the hemagglutinin was estimated at the micromolar level. Presented experimental data confirmed that both natural ORNs and functionalized ORNs-D-M inhibit the interactions between HA and glycan receptors and demonstrate anti-influenza activity.

Target of achieving exclusive breastfeeding is 33.6%, still low (IDHS 2010). Exclusive breastfeeding is intended to guarantee the fulfillment of the right of infants to 6 months old, with attention to growth and development, so that every mother would give exclusive breastfeeding to baby (PP no.23, 2012), the problem is that the milk does not come out in the first day of birth. It should be anticipated since pregnancies through lactation counseling, but the dissemination of information among the officers have not been optimized, so it was needed an alternative ways of stimulating milk production (Wiwin W, 2014). This study aims was to determine the SPEOS methods to increase milk production and baby weight.This research design used Quasi Experiment. Sample were primiparous postpartum mothers who were given the intervention SPEOS methods on the first day postpartum, were began 1-6 hours until the fourth week, then milk production measured before and after the intervention to see milk production and an increase in baby weight. The data analysed univariat analysis, Wilcoxon test bivariate analysis, multivariate analysis, and ANCOVA analysis to control confounding factors. This results showed the method SPEOS effect on milk production, average of 131.87 (p=0.00) and increased infant weight, average of 483.30 g (p 0.00). Age and the food consumed by the mother during the study (maternal nutrition) did not affect milk production, whereas IMD effect on milk production with 0.389 square r (p 0.04). SPEOS methods effect on milk production and an increase in baby weight on postpartum mother in Bengkulu City in BPM, 2016. Expected to all health workers can use the SPEOS method as a standard procedure in stimulate milk production to increase exclusive breastfeeding targets.

INTRODUCTION:Autoimmune reactions in Graves' disease (GD) occur not only in the thyroid gland, but also in the orbital connective tissue, eyelids, extraocular muscles. The occurrence of orbitopathy in the course of GD is influenced by environmental factors, e.g. cigarette smoking. OBJECTIVES:The aim of the study was to analyze the effect of cigarette smoking on the efficacy of activity of radioiodine(131I) therapy in patients with GD. We also studied the influence of cigarette smoking and the efficacy of prednisone prophylaxis on the risk of thyroid-associated ophthalmopathy (TAO) development after radioiodine therapy (RIT) during two years of follow-up. PATIENTS AND METHODS:Medical records of hyperthyroid patients treated with radioiodine had been included. Patients were scheduled to visit outpatient clinics at baseline and 1, 3, 6, 9, 12, 18, and 24 months after RIT. RESULTS:The studied group consisted of 336 patients (274 women, 62 men) diagnosed with GD and treated with RIT; 130 patients received second therapeutic dose of 131I due to recurrent hyperthyroidism. Among all studied patients, 220 (65.5%) were smokers and 116 (34.5%) non-smokers. In the group of smokers 115 (52.2%) of patients received single RIT, 105 (47.8%) received second dose of RAI due to recurrent hyperthyroidism. In non-smokers 91 (78.6%) received single activity of RAI, while 25 (21.4%) patients required second RIT due to recurrent hyperthyroidism. The ophthalmic symptoms in the group of smokers after RIT were less frequent, if the patient received preventative treatment in the form of oral prednisone (P = 0.0088). CONCLUSIONS:The results of our study suggest that cigarette smoking reduces the efficacy of treatment with 131I in patients with GD. The study also confirmed the effectiveness of steroid prophylaxis against TAO development or exacerbation after RIT.

The issue of increased risk of benign and malignant neoplasms in patients with acromegaly remains the topic of debate from many years and was addressed by numerous studies. Many of them have shown increase in the cancer incidence. Among particular types of malignancies, thyroid, colorectal, and breast cancer are most commonly indicated as associated with acromegaly. Single reports denoted increase in prevalence of neoplasms of other organs such as kidney, bone or central nervous system. Cardiovascular and respiratory tract disorders were traditionally consider as main causes of mortality in acromegalic patients, accounting for about 60 and 25% of deaths, respectively. However, according to a number of studies published over the current decade cancer became the most important cause of deaths. Aim of the current article was to review the literature concerning the risk of malignant neoplasms in acromegaly and its clinical implications.

Gastroenteropancreatic neuroendocrine tumors (GEP/NET) are unusual and relatively rare neoplasms that present many clinical challenges. They characteristically synthesize, store and secrete a variety of peptides and neuroamines which can lead to the development of distinct clinical syndromes, including the carcinoid syndrome, insulinoma, glucagonoma or VIPoma, although many are clinically silent until late presentation with mass effects. Management strategies include surgery and cytoreduction with chemotherapy and the use of somatostatin analogues to control symptoms from peptide neurotransmitter release. Somatostatin have a broad range of biological actions that include inhibition of exocrine and endocrine secretions, gut motility, cell proliferation, cell survival and angiogenesis. Five G-protein-coupled somatostatin receptors (SSTR1-SSTR5) have been cloned and characterized. Clinically used somatostatin analogues including octreotide and lanreotide are effective medical tools in treatment and show selectivity for SSTR2 and SSTR5. During treatment, disappearance of flushing, normalization of bowel movements and reduction of serotonin and 5-hydroxyindole acetic acid (5-HIAA) secretion are observed. The addition of interferon (INF) to somatostatin analogues may improve the antiproliferative effects. Patients who received the combination therapy had a significantly lower risk of progressive disease. On the other hand, the major toxicity of INF including temporary flu-like symptoms, tiredness, anorexia and leukopenia should be considered. Telotristat etiprate is an orally bioavailable, tryptophan hydroxylase (TPH) inhibitor with potential antiserotoninergic activity. Telotristat represents a novel approach by specifically inhibiting serotonin synthesis and, as such, is a promising potential new treatment for patients with carcinoid syndrome. Pancreatic functioning neuroendocrine tumors include insulinoma, gastrinoma, glucagonoma and VIPoma. Medical management in patients with insulinoma and severe hypoglycemic attacks includes diazoxide, which suppresses insulin release. Also mTOR inhibitors may inhibit insulin secretion. Treatment of gastrinoma includes both proton pump inhibitors (PPIs) and histamine H2-receptor antagonists. Octreotide and lanreotide can also control acid hypersecretion. In patients with glucagonomas hyperglycemia can be controlled using insulin and oral blood glucose lowering drugs. In malignant glucagonomas somatostatin analogues are effective in controlling necrolytic migratory erythema. Severe cases of the VIPoma syndrome require supplementation of fluid losses. Octreotide reduces tumoral VIP secretion and controls secretory diarrhea. Further control of diarrhea can be achieved with loperamide, opiates and glucocorticoids.

Studies revolving around mechanisms responsible for the development of amyloid-based diseases lay the foundations for the recognition of molecular targets of future to-be-developed treatments. However, the vast number of peptides and proteins known to be responsible for fibril formation, combined with their complexity and complexity of their interactions with various cellular components, renders this task extremely difficult and time-consuming. One of these proteins, human cystatin C (hCC), is a well-known and studied cysteine-protease inhibitor. While being a monomer in physiological conditions, under the necessary stimulus—usually a mutation, it tends to form fibrils, which later participate in the disease development. This process can potentially be regulated (in several ways) by many cellular components and it is being hypothesized that the cell membrane might play a key role in the oligomerization pathway. Studies involving cell membranes pose several difficulties; therefore, an alternative in the form of membrane mimetics is a very attractive solution. Here, we would like to present the first study on hCC oligomerization under the influence of phospholipid liposomes, acting as a membrane mimetic. The protein–mimetic interactions are studied utilizing circular dichroism, nuclear magnetic resonance, and size exclusion chromatography.

Glioblastoma multiforme (GBM) is the most common type of malignant gliomas, characterized by genetic instability, intratumoral histopathological variability and unpredictable clinical behavior. Disappointing results in the treatment of gliomas with surgery, radiation and chemotherapy have fueled a search for new therapeutic targets and treatment modalities. Here we report new approach towards RNA interference therapy of glioblastoma multiforme based on the magnetic nanoparticles delivery of the double-stranded RNA (dsRNA) with homological sequences to mRNA of tenascin-C (TN-C), named ATN-RNA. The obtained nanocomposite consisted of polyethyleneimine (PEI) coated magnetic nanoparticles conjugated to the dsRNA show high efficiency in ATN-RNA delivery, resulting not only in significant TN-C expression level suppressesion, but also impairing the tumor cells migration. Moreover, synthesized nanomaterials show high contrast properties in magnetic resonance imaging (MRI) and low cytotoxicity combining with lack of induction of interferon response. We believe that the present work is a successful combination of effective, functional, non-immunostimulatory dsRNA delivery system based on magnetic nanoparticles with high potential for further application in GBM therapy.

Polydopamine (PDA)-coated magnetic nanoparticles functionalized with mono-6-thio-β-cyclodextrin (SH-βCD) were obtained and characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), Nuclear and Magnetic Resonance Imaging (NMR and MRI), and doxorubicin (DOXO)-loading experiments. The liver cancer cellular internalization of DOXO-loaded nanoparticles was investigated by confocal imaging microscopy. Synthesized nanomaterials bearing a chemotherapeutic drug and a layer of polydopamine capable of absorbing near-infrared light show high performance in the combined chemo- and photothermal therapy (CT-PTT) of liver cancer due to the synergistic effect of both modalities as demonstrated in vitro. Moreover, our material exhibits improved T2 contrast properties, which have been verified using Carr-Purcell-Meiboom-Gill pulse sequence and MRI Spin-Echo imaging of the nanoparticles dispersed in the agarose gel phantoms. Therefore, the presented results cast new light on the preparation of polydopamine-based magnetic theranostic nanomaterials, as well as on the proper methodology for investigation of magnetic nanoparticles in high field MRI experiments. The prepared material is a robust theranostic nanoasystem with great potential in nanomedicine.

Thyroid nodular goiter is one of the most common medical conditions affecting even over a half of adult population. The risk of malignancy is rather small but noticeable-estimated by numerous studies to be about 3-10%. The definite differentiation between benign and malignant ones is a vital issue in endocrine practice. The aim of the current study was to assess the expression of vascular endothelial growth factor A (VEGF-A) and VEGF-C on the mRNA level in FNAB washouts in case of benign and malignant thyroid nodules and to evaluate the diagnostic value of these markers of malignancy.Patients undergoing fine-needle aspiration biopsy (FNAB) in our department between January 2013 and May 2014 were included. In case of all patients who gave the written consent, after ultrasonography (US) and fine-needle aspiration biopsy (FNAB) performed as routine medical procedure the needle was flushed with RNA Later solution, the washouts were frozen in -80 Celsius degrees. Expression of VEGF-A and VEGF-C and GADPH (reference gene) was assessed in washouts on the mRNA level using the real-time PCR technique. Probes of patients who underwent subsequent thyroidectomy and were diagnosed with differentiated thyroid cancer (DTC; proved by post-surgical histopathology) were analyzed. Similar number of patients with benign cytology were randomly selected to be a control group.Thirty one DTCs and 28 benign thyroid lesions were analyzed. Expression of VEGF-A was insignificantly higher in patients with DTCs (p = 0.13). Expression of VEGF-C was significantly higher in patients with DTC. The relative expression of VEGF-C (in comparison with GAPDH) was 0.0049 for DTCs and 0.00070 for benign lesions, medians - 0.0036 and 0.000024 respectively (p

BACKGROUND: In 2014, germline signal transducer and activator of transcription (STAT) 3 gain-of-function (GOF) mutations were first described to cause a novel multisystem disease of early-onset lymphoproliferation and autoimmunity. OBJECTIVE: This pivotal cohort study defines the scope, natural history, treatment, and overall survival of a large global cohort of patients with pathogenic STAT3 GOF variants. METHODS: We identified 191 patients from 33 countries with 72 unique mutations. Inclusion criteria included symptoms of immune dysregulation and a biochemically confirmed germline heterozygous GOF variant in STAT3. RESULTS: Overall survival was 88%, median age at onset of symptoms was 2.3 years, and median age at diagnosis was 12 years. Immune dysregulatory features were present in all patients: lymphoproliferation was the most common manifestation (73%); increased frequencies of double-negative (CD4-CD8-) T cells were found in 83% of patients tested. Autoimmune cytopenias were the second most common clinical manifestation (67%), followed by growth delay, enteropathy, skin disease, pulmonary disease, endocrinopathy, arthritis, autoimmune hepatitis, neurologic disease, vasculopathy, renal disease, and malignancy. Infections were reported in 72% of the cohort. A cellular and humoral immunodeficiency was observed in 37% and 51% of patients, respectively. Clinical symptoms dramatically improved in patients treated with JAK inhibitors, while a variety of other immunomodulatory treatment modalities were less efficacious. Thus far, 23 patients have undergone bone marrow transplantation, with a 62% survival rate. CONCLUSION: STAT3 GOF patients present with a wide array of immune-mediated disease including lymphoproliferation, autoimmune cytopenias, and multisystem autoimmunity. Patient care tends to be siloed, without a clear treatment strategy. Thus, early identification and prompt treatment implementation are lifesaving for STAT3 GOF syndrome.

PurposeA polygenic risk score (PRS) consisting of 313 common genetic variants (PRS313) is associated with risk of breast cancer and contralateral breast cancer. This study aimed to evaluate the association of the PRS313 with clinicopathologic characteristics of, and survival following, breast cancer.MethodsWomen with invasive breast cancer were included, 98,397 of European ancestry and 12,920 of Asian ancestry, from the Breast Cancer Association Consortium (BCAC), and 683 women from the European MINDACT trial. Associations between PRS313 and clinicopathologic characteristics, including the 70-gene signature for MINDACT, were evaluated using logistic regression analyses. Associations of PRS313 (continuous, per standard deviation) with overall survival (OS) and breast cancer-specific survival (BCSS) were evaluated with Cox regression, adjusted for clinicopathologic characteristics and treatment.ResultsThe PRS313 was associated with more favorable tumor characteristics. In BCAC, increasing PRS313 was associated with lower grade, hormone receptor-positive status, and smaller tumor size. In MINDACT, PRS313 was associated with a low risk 70-gene signature. In European women from BCAC, higher PRS313 was associated with better OS and BCSS: hazard ratio (HR) 0.96 (95% CI, 0.94 to 0.97) and 0.96 (95% CI, 0.94 to 0.98), but the association disappeared after adjustment for clinicopathologic characteristics (and treatment): OS HR, 1.01 (95% CI, 0.98 to 1.05) and BCSS HR, 1.02 (95% CI, 0.98 to 1.07). The results in MINDACT and Asian women from BCAC were consistent.ConclusionAn increased PRS313 is associated with favorable tumor characteristics, but is not independently associated with prognosis. Thus, PRS313 has no role in the clinical management of primary breast cancer at the time of diagnosis. Nevertheless, breast cancer mortality rates will be higher for women with higher PRS313 as increasing PRS313 is associated with an increased risk of disease. This information is crucial for modeling effective stratified screening programs.

We present a 3D model of the four transmembrane (TM) helical regions of bilitranslocase (BTL), a structurally uncharacterized protein that transports organic anions across the cell membrane. The model was computed by considering helix-helix interactions as primary constraints, using Monte Carlo simulations. The interactions between the TM2 and TM3 segments have been confirmed by Förster resonance energy transfer (FRET) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy, increasing our confidence in the model. Several insights into the BTL transport mechanism were obtained by analyzing the model. For example, the observed cis-trans Leu-Pro peptide bond isomerization in the TM3 fragment may indicate a key conformational change during anion transport by BTL. Our structural model of BTL may facilitate further studies, including drug discovery.

BackgroundReproductive factors have been shown to be differentially associated with risk of estrogen receptor (ER)-positive and ER-negative breast cancer. However, their associations with intrinsic-like subtypes are less clear.MethodsAnalyses included up to 23 353 cases and 71 072 controls pooled from 31 population-based case-control or cohort studies in the Breast Cancer Association Consortium across 16 countries on 4 continents. Polytomous logistic regression was used to estimate the association between reproductive factors and risk of breast cancer by intrinsic-like subtypes (luminal A-like, luminal B-like, luminal B-HER2-like, HER2-enriched-like, and triple-negative breast cancer) and by invasiveness. All statistical tests were 2-sided.ResultsCompared with nulliparous women, parous women had a lower risk of luminal A-like, luminal B-like, luminal B-HER2-like, and HER2-enriched-like disease. This association was apparent only after approximately 10 years since last birth and became stronger with increasing time (odds ratio [OR] = 0.59, 95% confidence interval [CI] = 0.49 to 0.71; and OR = 0.36, 95% CI = 0.28 to 0.46 for multiparous women with luminal A-like tumors 20 to less than 25 years after last birth and 45 to less than 50 years after last birth, respectively). In contrast, parous women had a higher risk of triple-negative breast cancer right after their last birth (for multiparous women: OR = 3.12, 95% CI = 2.02 to 4.83) that was attenuated with time but persisted for decades (OR = 1.03, 95% CI = 0.79 to 1.34, for multiparous women 25 to less than 30 years after last birth). Older age at first birth (Pheterogeneity < .001 for triple-negative compared with luminal A-like breast cancer) and breastfeeding (Pheterogeneity < .001 for triple-negative compared with luminal A-like breast cancer) were associated with lower risk of triple-negative breast cancer but not with other disease subtypes. Younger age at menarche was associated with higher risk of all subtypes; older age at menopause was associated with higher risk of luminal A-like but not triple-negative breast cancer. Associations for in situ tumors were similar to luminal A-like.ConclusionsThis large and comprehensive study demonstrates a distinct reproductive risk factor profile for triple-negative breast cancer compared with other subtypes, with implications for the understanding of disease etiology and risk prediction.

IntroductionIn the last decade, (18)F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET and PET/CT) has become one of the major diagnostic tools used in oncology. A significant number of patients who undergo this procedure, due to non-thyroidal reasons, present incidental uptake of (18F-FDG) in the thyroid. The aim of the study was to compare the SUVmax (standardized uptake value) of thyroid focal lesions, which were incidentally found on PET/CT, in relation to the results of thyroid fine-needle aspiration biopsy (FNAB) and/or histopathological evaluation.Materials and methodsPatients referred for PET/CT examination, due to non-thyroidal illness, presented focal 18F-FDG uptake in the thyroid and were advised to undergo ultrasonography (US), hormonal evaluation, FNAB and/or total thyroidectomy at our institution.Results6614 PET/CT examinations performed in 5520 patients were analyzed. Of the 122 patients with focal thyroid 18F-FDG activity, 82 patients (67.2%) underwent further thyroid evaluation using FNAB. Benign lesions were diagnosed in 46 patients, malignant - in 19 patients (confirmed by post-surgical histopathology), while 17 patients had inconclusive results of cytological assessment. Mean SUVmax of benign lesions was 3.2±2.8 (median = 2.4), while the mean SUVmax value for malignant lesions was 7.1±8.2 (median = 3.5). The risk of malignancy was 16.7% for lesions with a SUVmax under 3, 43.8% for lesions with a SUVmax between 3 and 6, and 54.6% for lesions with a SUVmax over 6. In the group of malignant lesions, a positive correlation between the lesion's diameter and SUVmax was observed (p = 0.03, r = 0.57).ConclusionsSubjects with incidental focal uptake of 18F-FDG in thyroid are at a high risk of thyroid malignancy. A high value of SUVmax further increases the risk of malignancy, indicating the necessity for further cytological or histological evaluation. However, as SUVmax correlated with the diameter of malignant lesions, small lesions with focal uptake of 18F-FDG should be interpreted cautiously.

INTRODUCTION: Acromegaly is a quite rare chronic disease caused by the increased secretion of growth hormone (GH) and subsequently insulin - like growth factor 1. Although cardiovascular diseases remains the most common cause of mortality among acromegalic patients, increased prevalence of malignant and benign neoplasms remains a matter of debate. The aim of this study is to evaluate the risk of thyroid nodular disease (TND) and thyroid cancer in patients with acromegaly. MATERIALS AND METHODS: PubMed, Cochrane Library, Scopus, Cinahl, Academic Search Complete, Web of Knowledge, PubMed Central, PubMed Central Canada and Clinical Key databases were searched to identify studies containing. Random-effects model was used to calculate pooled odds ratios and risk ratios of TND in acromegaly. Studies which not included control groups were systematically reviewed. RESULTS: TND was more frequent in acromegaly than in control groups (OR = 6.9, RR = 2.1). The pooled prevalence of TND was 59.2%. Also thyroid cancer (TC) proved to be more common in acromegalic patients (OR = 7.5, RR = 7.2), prevalence was 4.3%. The pooled rate of malignancy (calculated per patient) was equal to 8.7%. CONCLUSIONS: This study confirms that both TND and TC occur significantly more often in acromegalic patients than in general population. These results indicate that periodic thyroid ultrasound examination and careful evaluation of eventual lesions should be an important part of follow-up of patients with acromegaly.

IntroductionThyroid nodular disease (TND) is a very common disorder. However, since the rate of malignancy is reported to be 3-10%, only a minority of patients require aggressive surgical treatment. As a result, there is a need for diagnostic tools which would allow for a reliable differentiation between benign and malignant nodules. Although a number of conventional ultrasonographic (US) features are proved to be markers of malignancy, Shear Wave Elastography (SWE) is considered to be an improvement of conventional US. The aim of this study was to compare conventional US markers and SWE diagnostic values in the differentiation of benign and malignant thyroid nodules.Materials and methodsAll patients referred for thyroidectomy, irrespective of the indications, underwent a US thyroid examination prospectively. Patients with TND were included into the study. Results of the US and SWE examinations were compared with post-surgical histopathology.ResultsOne hundred and twenty two patients with 393 thyroid nodules were included into the study. Twenty two patients were diagnosed with cancer. SWE turned out to be a predictor of malignancy superior to any other conventional US markers (OR=54.5 using qualitative scales and 40.8 using quantitative data on maximal stiffness with a threshold of 50 kPa).ConclusionsAlthough most conventional US markers of malignancy prove to be significant, none of them are characterized by both high sensitivity and specificity. SWE seems to be an important step forward, allowing for a more reliable distinction of benign and malignant thyroid nodules. Our study, assessing SWE properties on the highest number of thyroid lesions at the time of publication, confirms the high diagnostic value of this technique. It also indicates that a quantitative evaluation of thyroid lesions is not superior to simpler qualitative methods.

Ethiopian mustard (Brassica carinata) is an ancient crop with remarkable stress resilience and a desirable seed fatty acid profile for biofuel uses. Brassica carinata is one of six Brassica species that share three major genomes from three diploid species (AA, BB, and CC) that spontaneously hybridized in a pairwise manner to form three allotetraploid species (AABB, AACC, and BBCC). Of the genomes of these species, that of B. carinata is the least understood. Here, we report a chromosome scale 1.31-Gbp genome assembly with 156.9-fold sequencing coverage for B. carinata, completing the reference genomes comprising the classic Triangle of U, a classical theory of the evolutionary relationships among these six species. Our assembly provides insights into the hybridization event that led to the current B. carinata genome and the genomic features that gave rise to the superior agronomic traits of B. carinata. Notably, we identified an expansion of transcription factor networks and agronomically important gene families. Completion of the Triangle of U comparative genomics platform has allowed us to examine the dynamics of polyploid evolution and the role of subgenome dominance in the domestication and continuing agronomic improvement of B. carinata and other Brassica species.

Germline copy number variants (CNVs) are pervasive in the human genome but potential disease associations with rare CNVs have not been comprehensively assessed in large datasets. We analysed rare CNVs in genes and non-coding regions for 86,788 breast cancer cases and 76,122 controls of European ancestry with genome-wide array data. Gene burden tests detected the strongest association for deletions in BRCA1 (P = 3.7E-18). Nine other genes were associated with a p-value