Heterologous Expression of Interferon α-2b in Lactococcus lactis and its Biological Activity against Colorectal Cancer Cells

( Lita Meilina ),( Sri Budiarti ),( Apon Zaenal Mustopa ),( Huda Shalahudin Darusman ),( Lita Triratna ),( Muhammad Ajietuta Nugraha ),( Muhammad Sabiq Bilhaq ),( Ratih Asmana Ningrum ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 한국미생물·생명공학회지 Vol.49 No.1

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SPU_sp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 μg/ml and 127.2 μg/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 μg/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 μg/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

Developmental and Transfer Issue of L2 Pragmatic Knowledge

Lita M. de Guzman 한국중원언어학회 2018 언어학연구 Vol.0 No.49

The present study investigates the development of request realization of Tagalog learners of English and the transfer of learners’ L1 pragmatic knowledge to the target language when learners make requests. The data were collected through written discourse completion task from three different English proficiency levels in six situations and compared against native speakers of English and native speakers of Tagalog. Statistical analyses reveal that as L2 learners become more proficient at L2, they demonstrate more positive transfer in their use of external modifiers and negative transfer in internal modifiers. L2 learners were found to differ from English native speakers with respect to perception of situational variations such as power and social familiarity. However, it was also found that all learner groups and control groups were equally sensitive to situational variations in the sense that they varied their uses of internal and external modifications as the situations varied. With respect to developmental stages in pragmatic development, learners’ overall proficiency cannot reliably predict interlanguage pragmatic development, which implies that there is a mismatch between a learner’s linguistic competence and pragmatic competence.

Ceramide’s Role and Biosynthesis: A Brief Review

Lita Amalia,Shen-Long Tsai 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.3

The utilization of ceramides, which are members of the sphingolipid family, has been widely acknowledged in the cosmetic and pharmaceutical industries, along with various other applications as therapeutic agents. Most ceramides currently available on the market are synthetic ceramides created through chemical reactions with precursors resembling the natural precursor of sphingolipid production by living organisms. In fact, many organisms ranging from microbes to higher-order mammals natively use metabolism to produce sphingolipids, including ceramides and their derivatives, to support cell molecular functions. Sphingolipids, for instance, are present in the cell membranes of mammals, plants, and yeast to maintain membrane morphology. As a green alternative to the chemical synthesis method, many studies have been carried out to reveal the diversity and characteristics of biologically produced ceramide derivatives. In this review, we summarized the most important aspects of ceramide biosynthesis in general and provide a quick overview of the common organisms producing ceramides. In addition, a brief discussion regarding the role of ceramides in cells and their risks was included. As the biosynthesis of ceramides is an attractive alternative to current commercial methods, the advances reviewed herein demonstrate the untapped potential for the further development of synthetic pathways to enhance biobased-ceramide production.

On the rate of convergence of uniform approximations for sequences of distribution functions

João Lita da Silva,Luís Pedro Ramos 한국통계학회 2014 Journal of the Korean Statistical Society Vol.43 No.1

In this paper, we develop uniform bounds for the sequence of distribution functions ofg(Vn + μn), where g is some smooth function, {Vn, n ≥ 1} is a sequence of identicallydistributed random variables with common distribution having a bounded derivative and{μn} are constants such that μn → ∞. These bounds allow us to identify a suitablesequence of random variables which is asymptotically of the same type of g(Vn + μn)showing that the rate of convergence for these uniform approximations depends on theratio of the second derivative to the first derivative of g. The corresponding generalizationto the multivariate case is also analyzed. An application of our results to the STATIS-ACTmethod is provided in the final section.

Health risk assessment of pest control and disinfection workers before and after the COVID-19 outbreak in South Korea

Yun-Hee Choi(Yun-Hee Choi),Lita Kim(Lita Kim),Sang ji Lee(Sang ji Lee),Da-An Huh(Da-An Huh),Kyong Whan Moon(Kyong Whan Moon) 한국환경보건학회 2022 한국환경보건학회 학술대회 자료집 Vol.2022 No.5

Spatial ability and anatomy learning performance among dental students

Sarilita Erli,Lita Yurika Ambar,Firman Dani Rizali,Wilkinson Tracey,Susilawati Sri,Saptarini Risti,Aripin Dudi,Sjamsudin Endang 한국의학교육학회 2022 Korean journal of medical education Vol.34 No.4

Purpose: Spatial perception is an essential skill for professional dentists. The objective of this study was to observe the spatial ability, as well as anatomy module grades, of dental students at a dental education center in Indonesia and relate these to gender and cohort.Methods: A cross-sectional study was carried out where dental students in years (cohorts) 1, 2, 4–6 were invited to take the Revised Purdue Spatial Visualization Test (PSVT-R) and the redrawn Vandenberg and Kuse Mental Rotation Test (MRT) in order to assess spatial ability. In addition, the 1st- and 2nd-year dental students carried out gross anatomy assessments. Spatial ability test results were compared using an independent t-test to detect gender differences, one-way analysis of variance to inspect cohort differences, and correlation relative to anatomy module scores.Results: A total of 326 dental students voluntarily participated. Statistically significant gender differences were found in both spatial ability tests in the overall sample (PSVT-R: p<0.001; MRT: p=0.001). When the 1st- and 2nd-year dental students were pooled, significant gender differences were detected, in which males scored higher than females in both spatial ability tests (PSVT-R: p<0.001; MRT: p=0.003). In anatomy, however, females scored higher than the males (p=0.005). In addition, there were weak to moderate, but significant correlations between spatial ability tests and anatomy scores.Conclusion: This study indicated that spatial ability may not be the only factor predicting the academic performance of dental students. However, dental students with low spatial ability scores may need supplementary educational techniques when learning specific spatial tasks.

Volumetric growth analysis of maxillary sinus using computed tomography scan segmentation: a pilot study of Indonesian population

Erli Sarilita,Yurika Ambar Lita,Harry Galuh Nugraha,Nani Murniati,Harmas Yazid Yusuf 대한해부학회 2021 Anatomy & Cell Biology Vol.54 No.4

The aim of this study is to investigate the volumetric measurements of the maxillary sinus among Indonesian population through computed tomography (CT) scan semi-automated segmentation. This project collected 802 retrospective head CT scan archives from Department of Radiology, Hasan Sadikin Hospital, Bandung, Indonesia between 2019-2020. Patients with craniofacial anomalies/pathology fracture in proximity of the maxillary sinuses, and mediocre image quality were excluded from this study resulting only 97 CT scan archives (194 maxillary sinuses; 52 males; 45 females; age range 0-25 years old). Three-dimensional craniofacial structures were reconstructed and volumetric measurements of the maxillary sinus were computed through semi-automated segmentation using ITK-SNAP. This study recorded the initial phase of maxillary sinus pneumatization during infancy. The maxillary sinus developed until reaching the maximum of average maxillary sinus volume at 13,278.73 mm3 in 16 to 20 years old group in which afterwards fell to 12,325.21 mm3. There was no difference found between right and left maxillary sinus volume. This study revealed that the pneumatization of maxillary sinus begin during infancy and climb until reaching the second decade of life, in which after that slowly decrease. Moreover, no difference between right and left maxillary sinus volume was detected. The volumetric dimension of maxillary sinus presented in this study may serve as the basis knowledge surgical intervention of maxillary sinus and its related structures.

Work-Rest Schedules for Hand Tractor Field Operation with Hand-Transmitted Vibration Exposure Limitation

Ubaidillah Ubaidillah,Lutfi Musthofa,Darmanto Darmanto,Perdana Lita Puspita R. 한국농업기계학회 2023 바이오시스템공학 Vol.48 No.4

Purpose The hand tractors produce hand-transmitted vibration, a significant occupational hazard for hand tractor operators. The present study aims to characterize the hand-transmitted vibration of hand tractors under various engine rotational speeds (1400 rpm, 1700 rpm, and 2000 rpm) and soil implements (moldboard plow, rotary harrow, and leveler) and develop work-rest schedules for hand tractor operations. Methods The study involved field experiments in soil wet preparation and soil dry preparation operations to measure the vibration acceleration at the handle grip and the operator’s physiological cost of work. The data was then analyzed to obtain the maximum allowable time based on the American Conference of Governmental Industrial Hygienists (ACGIH) limits and the National Institute for Occupational Safety and Health (NIOSH) calculations. Results The results showed that vibration acceleration was significantly affected by engine rotational speed and the soil implement used. The vibration acceleration was highest on the x-axis, followed by the z- and y-axis. The maximum allowable time was 3 h/day/operator and 2.5 h/day/operator and divided into four and five work bouts with 45 min and 30 min each for wet and dry soil preparation, respectively. The heart rate data indicated that at least 15 min of rest should be given in between work bouts, with an additional 30 min and 15 min of free time for each wet and dry soil operation, respectively. Conclusions Based on this research, a work-rest schedule was designed for two hand tractor operators who work alternately to minimize hand-arm vibration syndrome.

The Expression of Codon Optimised Hepatitis B Core Antigen (HBcAg) of Subgenotype B3 Open Reading Frame in Lactococcus lactis

( Apon Zaenal Mustopa ),( Sri Kartika Wijaya ),( Ratih Asmana Ningrum ),( Dian Fitria Agustiyanti ),( Lita Triratna ),( Wida Nurul Alfisyahrin ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 한국미생물·생명공학회지 Vol.47 No.3

Hepatitis B treatments using immune therapy are gaining interest because of the improvements in dendritic cell performance for antigen presentation, which induces an appropriate immune response and raises patient survival rates. This research aims to produce a significant amount of the HBcAg antigen, which can induce an immune response and have a curative effect on HBV infection. In this study, the HBV subgenotype B3 of the HBcAg gene was used, which is dominant in Indonesia. Further, Lactococcus lactis bacteria was used as the host because of its safety and tightly regulated protein expression. The codon usage for the HBcAg gene was optimized to improve protein expression in L. lactis, which is important because a codon is not random between species. The HBcAg gene is attached to a pNZ8148 plasmid and transformed into the L. lactis NZ3900 expression host. The results confirm that a positive protein band (21 kDa) in two fractions of purified HBcAg was recognized by both western blotting and dot blot hybridization, even if the HBcAg optimized codon has higher GC contents than that suggested for L. lactis expression. Overall, this research strengthens the broad use of L. lactis bacteria for any protein expression, including higher protein expression of codon optimized HBcAg gene compared to non-optimized genes. Furthermore, the improvement in the codon optimization of the HBcAg gene significantly increases the total protein expression by 10-20%, and the expression level of the codon optimized HBcAg increases 1.5 to 3.2-times that of the native HBcAg.

Volumetric analysis of normal condyles and those with disc displacement with reduction in the Indonesian population: A CBCT study

Nawawi Azkya Patria,Rikmasari Rasmi,Kurnikasari Erna,Oscandar Fahmi,Lita Yurika Ambar 대한영상치의학회 2022 Imaging Science in Dentistry Vol.52 No.1

Purpose: Disc displacement can cause resorption of the head of the condyle and affect its volume. This study analysed the volume of normal condyles and those with disc displacement with reduction (DDR) in cone-beam computed tomography (CBCT) scans from the Indonesian population. Materials and Methods: This study analysed 56 condyles (26 normal and 30 with DDR) from patients who visited the Oral and Maxillofacial Radiology Unit after being referred from the Prosthodontics Unit at Dental Hospital Universitas Padjadjaran from December 2020 to February 2021. Samples were divided into 2 groups (normal and DDR left and right-side condyles) based on the DC/TMD Axis 1 form through the clinical examination results. Both sample groups were exposed to CBCT radiation. The CBCT imaging results in the Digital Imaging and Communications in Medicine format were exported to the open-source ITK-SNAP format to determine condyle volume. Volumetric data from the cortical and trabecular areas of the right or left side condyles were arranged by sex. The independent t-test was used to determine the significance of differences with IBM SPSS version 21.0. Intra- and inter-observer reliability and validity were tested before determining the volume of the condyles. Results: Normal condyles and DDR condyles showed significant differences in volume (P<0.05). Significant differences were also seen in cortical (P = 0.0007) and trabecular (P = 0.0045) volumes. There was a significant difference in condylar volume based on sex. Conclusion: The normal condyle volume was significantly different from the DDR condyle volume in both sexes.