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Heterologous expression of proteins often pursues high expression levels, but it can easily result in misfolding and loss of biological function. L-α-glycerophosphate oxidase (GlpO) is a flavin adenine dinucleotide (FAD)-dependent oxidase which is widely used in the clinical determination of triglycerides. We found that the total enzymatic activity of GlpO expressed in Escherichia coli (E. coli) was extremely low, probably due to the absence of FAD cofactors and the misfolding of GlpO at a high synthesis rate. Therefore, decreasing the expression rate was used to improve the activity of GlpO. The specific activity of GlpO expressed on the pUC19 vector with lac promotor was approximately 30 times higher than that expressed on the pET28a vector with T7 promotor, but the expression levels of GlpO on the two vectors were completely opposite. It indicated that the specific activity of GlpO was increased as the expression level decreased. However, too low expression greatly influences the total amount and activity of the functional enzyme. In order to resolve this problem, two new plasmids, GlpO-CG4 and GlpO-CG6, were constructed by inserting 4 or 6 nucleotides, respectively, between the ribosome binding site (RBS) and the start code (ATG) on pET28a. Compared with the expression on the GlpO-pET vector, the expression rates of GlpO on the GlpO-CG4 and GlpO-CG6 were dramatically decreased. The total activity of GlpO expressed on GlpO-CG6 was 11 times and 1.5 times higher than that expressed on the GlpO-pET and GlpO-pUC, respectively. Results suggest that the activity of GlpO can be improved by decreasing the expression rate. 

Calcinosis cutis (CC) [1] is an unusual disorder characterized by calcium-phosphate deposition into cutaneous and subcutaneous tissues. There are five subtypes: dystrophic, metastatic, idiopathic, iatrogenic and calciphylaxis.Calciphylaxis or calcifying panniculitis is defined as small vessel calcification mainly affecting blood vessels of the dermis and subcutaneous fat. Despite the predominance of cases in patients with ESRD, calciphylaxis can also be found in patients with normal renal function and normal levels of calcium and phosphate. These cases are often referred to as nonuremic calciphylaxis (NUC), a heterogeneous category with several associations. Literature reveals an association with hyperparathyroidism (28%), malignancy (22%), alcoholic liver disease (17%) and connective tissue diseases (11%) while obesity, liver disease, high-serum calcium (Ca) × phosphorus (P) levels, combined therapies of calcium salts with vitamin D, warfarin and corticosteroids have been observed to increase the likelihood of this disease [2]. The lesions in both nonuremic and uremic calciphylaxis tend to be indistinguishable from each other, initially presenting as tender subcutaneous plaques that progress into nonhealing ulcers with overlying black eschar. Skin changes often begin with a livedo reticularis pattern that can progress to livedo racemes and ultimately retiform purpura.In our clinical case, we describe a patient with multiple risk factors for calciphylaxis, intense widespread calcification (vessels, tendons, joints) and cutaneous calcific stone of calcium and phosphate oxalate not elsewhere described before.

Clinical applications of Artificial Intelligence (AI) in healthcare are relatively rare. The high expectations in relation to data analysis influencing general healthcare have not materialized, with few exceptions, and then predominantly in the field of rare diseases, oncology and pathology, and interpretation of laboratory results. While electronic health records, introduced over the last decade or so in the UK have increased access to medical and treatment histories of patients, diagnoses, medications, treatment plans, immunization dates, allergies, radiology images, laboratory and test results, these have potential for evidence-based tools that providers can use to make decisions about a patient’s care, as well as streamline workflow. In the following text, we review the advances achieved using machine learning and deep learning technology, as well as robot use and telemedicine in the healthcare of older people. Key points: 1. Artificial Intelligence use is extensively explored in prevention, diagnosis, novel drug designs and after-care. 2. AI studies on older adults include a small number of patients and lack reproducibility needed for their wider clinical use in different clinical settings and larger populations. 3. Telemedicine and robot assisted technology are well received by older service users. 4. Ethical concerns need to be resolved prior to wider AI use in routine clinical setting.

The study on the remote effect of fascial manipulation on knee pain presents a case report of a patient with knee pain who underwent a single session of fascial manipulation to reduce pain and improve daily activities. The report explains the anatomy of the knee joint and the potential causes of knee pain, including bony and soft tissue involvement. Fascial manipulation is a technique that involves applying mechanical force to areas of fascial dysfunction to stimulate the body’s natural inflammatory response. The therapy is based on applying physical friction to the densified connective tissue with an elbow or knuckle to raise the temperature and change the consistency of loose connective tissue rich in hyaluronic acid. The locations picked were proximal and distal to the problematic area, and no point was treated over the painful region, which is consistent with the notion of pain being caused by aberrant fascial tensions. Manipulation of the connective tissue is thought to cause mechanical stress and heat, resulting in less macromolecular crowding and defragmentation of the Hyaluronic Acid (HA) polymers and normalised fascial gliding across the connective tissue. Reduced discomfort and enhanced range of motion are two more regularly reported effects of Fascial Manipulation. The patient was urged to resume his everyday activities after each treatment session to favour typical physiological motions that would align collagen fibres along normal lines of force. The report concludes that fascial manipulation can be a beneficial technique for managing knee pain, and it can be complemented with exercises and stretches to improve outcomes.

Background: Stillbirth (SB), either because of intrauterine or intrapartum fetal death, is amongst the most devastating pregnancy complications, hardest to predict, real challenge for health systems. India probably accounts for the highest SB numbers in the world.Objectives: A community-based prospective study was carried out to know the burden, and causes of SBs in rural women of remote region.Material methods: After approval of the ethics committee of the Institute in Maharashtra for broader research, which was service-oriented, the present study was conducted in 100 villages around the village with health facilities. These villages were included keeping in mind future services. After obtaining the consent of women, information was prospectively recorded on their predesigned tool (which was for broader work), by asking women and seeing their records of antenatal and intranatal care on a regular basis. The study subjects were selected after obtaining information from nurse midwives, Accredited Social Health Activists (ASHAs), and Aaganwadi workers. As the plan was to serve, there was excellent cooperation. The tool was made for broader work. Villages were visited 5 days a week. Study was community-based in villages in remote, hilly region with various local issues so any information the women gave conclusion and was in the record was used.Results: A total of 3905 births occurred over two years, 3635 (93%) term, and 270 (7%) preterm. Amongst 3635 term births, 3474 (95.6%) were live births, 161 (4.4%) SBs, amongst 270 preterm births, 239 (88.5%) were live births, 31 (11.5%) SBs. Of 192 SBs, 48 (7.2%) SBs were amongst 2690 women with anaemia, (16 (2.4%) SBs amongst women with very severe anaemia, 12 (1.8%) SBs were amongst 667 women with severe anaemia, 12 (1.8%) SBs were amongst 664 women with moderate anaemia, 8 (1.2%) SBs were amongst 680 women with mild anaemia). Overall of 192 (4.9%) SBs, 31 (18.1%) SBs were among 517 women with hypertensive disorders of pregnancy (HDsP) 8 (4.4%) SBs amongst 280 women with moderate HDsP, 12 (6.9%) SBs amongst 175 women with severe HDsP, 11 (6.8%) SBs in 62 women with eclampsia. Overall of 192 SBs, 6 (3.1%) babies had weight < 1 kg, 8 (4.1%) of ≥ 1 to < 1.5 kg, 50 (26.0%) ≥ 1.5 to < 2 kg, 64 (33.3%) ≥ 2 to < 2.5 kg, 64 (33.3%) 2.5 kg and more, obviously fewer babies more deaths in birth weight less than 1. 5 kg, more so less than 1 kg.Conclusion: Overall SBs were in women with disorders but 4.5% SBs did occur in women with no disorder. SBs were significantly higher amongst babies weighing less than 1 kg, beyond this weight, numbers were similar.

This paper is an attempt to enrich the literature about the role that can play some economic approaches, such as cost-effectiveness analysis, in order to help medical stuffs to decide about the treatment to adopt in case of a chronic disease such as Haemophilia. Data from morocco were gathered in order to explain the importance of such approaches.

Coronavirus disease 2019 (COVID-19) which first appeared in China spread gradually all over the world within three months [1]. China was the only country mainly affected by Covid-19 until February 2020, but from the beginning of March, the disease started to spread rapidly to South Korea. It reached Italy in the second week of March and the number of cases increased rapidly in Spain and other European countries in the third week of March then the virus crossed the Atlantic and entered into the United States and other countries in the Americas. WHO declared COVID-19 as a pandemic disease on 11th March 2020 [2]. As of 23rd April 2020, there have been 2,645,785 confirmed cases of COVID-19, with 185,121 deaths and 726,827 recoveries [3]. Slowly, Nepal is also into the scene of the COVID-19 affected countries.

We demonstrated intrinsic connections between left ventricular hypertrophy (LVH) and arterial hypertension (AHT) through the recently announced centralized aerobic-anaerobic energy balance compensation (CAAEBC) theory. CAAEBC has already demonstrated achievements in the treatment of AHT, diabetes myelitis (DM), and osteochondrosis. Such demonstration lifts the necessity to check the applicability of this theory to other non-communicable diseases (NCDs) and develop the proper way to model the main idea of CAAEBC.

Premature ovarian insufficiency (POI) is a rare disease, especially in children and adolescents. It was previously called premature ovarian failure (POF). It can be manifested as delayed puberty, primary or secondary amenorrhea that occurred before the age of 40 years with no less than two abnormal serum sex hormones (low estrogen and high gonadotropin). It is reported that the incidence rate is 1% at the age of 40 years and 0.01% at the age of 20 years. Although the disease usually occurs in middle-aged and elderly women, clinical practice in recent years has shown that it has also been found in adolescents and even children. It is generally believed that the etiology of POI includes genetic factors, immune factors, and iatrogenic factors. So far, several genetic mutations that may cause POI have been found clinically, but the etiology of 90% of POI is still unknown. In recent years, the incidence of POI in children and adolescents has increased, and there are more urgent requirements for its early diagnosis, treatment, and clinical management. Based on this, this article will mainly review the research progress of the etiology, treatment, and clinical management of POI in children and adolescents.

Background: School nurses possess an essential role in treating and helping children maintain health. However, the full scope of their role has not been identified by parents. Therefore, the purpose of this study was to explore parents’ perceptions of the role of the school nurse. Methods: Descriptive statistics were used to analyze the data. One hundred eighty parents participated in the study. The perception of the roles of school nurses was assessed by a 16-item questionnaire, which was adapted from a study by Kirchofer, et al. 2007. Results: The four most important school nurses’ roles perceived by the parents were providing first aid and emergency care to children, communicating with parents and health care providers in the presence of a problem, providing medical treatment, and preventing and controlling diseases. Conclusion: School nurses have many important roles, and while parents identified some essential roles, they did not recognize other vital roles as being very important. Increasing awareness of school nurses’ multifaceted roles among parents is essential so that they can utilize nurses’ expertise in maintaining their student’s health as well as tap into a key resource in the coordination of care for their child.