Medical Biotechnology

Human pluripotent stem cells (hPSCs) have the potential to revolutionize cell-replacement therapies because of their ability to self renew and differentiate into nearly every cell type in the body. However, safety concerns have delayed the clinical translation of this technology. One cause for this is the capacity that hPSCs have to generate tumors after transplantation. Because of the challenges associated with achieving complete differentiation into clinically relevant cell types, the development of safe and efficient strategies for purifying committed cells is essential for advancing hPSC-based therapies. Several purification strategies have now succeeded in generating non-tumorigenic and homogeneous cell-populations. These techniques typically enrich for cells by either depleting early committed populations from teratoma-initiating hPSCs or by positively selecting cells after differentiation. Here we review the working principles behind separation methods that have facilitated t...

Background: Excessive continuous NO release from inducible NO synthase over prolonged periods under pathological conditions, such as endotoxemia, contributes significantly to circulatory failure, hypotension, and septic shock. This NO production during endotoxemia is accompanied by superoxide release, which contributes to the fast decay of NO. Therefore, the amount of NO that diffuses to target sites may be much lower than the total amount released under pathological conditions. Methods: We performed in vivo and ex vivo measurements of NO (electrochemical) and ex vivo in situ measurements of superoxide, peroxynitrite (chemiluminescence), and nitrite and nitrate (ultraviolet-visible spectroscopy). We determined the effect of lipopolysaccharide administration (20 mg/kg) on diffusible NO, total NO (diffusible plus consumed in chemical reactions), and superoxide and peroxynitrite release in the pulmonary arteries of rats. Results: An increase in diffusible NO generated by constitutive N...

As health care centres have becomes popular, daily monitoring of health-status related parameters is becoming important. An easy, comfortable and patient friendly solution for acquisition, processing and remotely transmitting the information from patient to the centre is therefore an important issue. Phonocardiogram (PCG) is a physiological signal reflecting the cardiovascular status. This paper deals with a Signal Processing Module for the computer-aided analysis of the condition of heart. The module has three main blocks: Data Acquisition, Signal Processing & Remote Monitoring of heart sounds. Data acquired includes the heart sounds. The system integrates embedded internet technology and wireless technology. As the data is being send by internet, it realizes real-time recording and monitoring of physiological parameter of patients at low cost and both at home and in hospital. The analysis can be carried out using computer initially and further by doctor. The tele-monitoring system...

In order to improve the quality of medical student training, the clinical case-based learning (CBL) teaching mode is analyzed in the aspect of the rate of achievement, practical ability, and psychological effect. A total of 86 medical students who came to our hospital from March 2020 to December 2021 are selected and divided into the CBL group and the control group according to the random number table method. The psychological quality of medical students is evaluated by observing their movements and expressions during their internship. In addition, the questionnaire is used to evaluate the scores of a medical student in future planning, occupational health, and teamwork spirit. The compliance evaluation results show that CBL teaching mode can significantly improve the compliance rate of medical students in training and promote the confidence of future role change to a certain extent.

Carbonic anhydrase VI (CA VI) is a secretory isoenzyme that, by analogy to α-amylase, is produced in the salivary glands and delivered into saliva. To determine whether CA VI is transferred into the circulation and is detectable in human serum, we collected blood samples from four healthy subjects at 3-h intervals throughout a 24-h period and measured concentrations of CA VI by a specific time-resolved immunofluorometric assay. All serum samples contained CA VI, the concentrations being ∼22 times lower in serum than in the corresponding saliva samples. The presence of CA VI in serum was confirmed by Western blotting, which under reducing conditions identified a 42-kDa polypeptide band corresponding to the monomeric CA VI. The described time-resolved immunofluorometric assay for CA VI might be useful to identify or exclude diseases of the salivary glands in the differential diagnosis of patients whose serum amylase concentrations are increased.

In this HPLC–diode-array detection method for toxicological drug screening, a mixed-mode solid-phase extraction procedure is optimized for isolation of a broad range of drugs from serum and urine. Basic, neutral, and weakly acidic drugs are uniformly recovered. The extract from the solid-phase cartridge is readily injected to a reversed-phase HPLC column for separation by gradient elution. Unknown drugs and metabolites in urine and serum samples from acute drug poisoning cases are rapidly identified by matching their retention times and ultraviolet spectra with hundreds of reference compounds in the library. Urine metabolites of common toxicants from various medications and drugs of abuse are recorded, with their changes of retention times and ultraviolet spectra as related to their metabolic transformations. Glucuronide conjugates of common benzodiazepines, tricyclic antidepressants, and beta-blockers are examined directly without chemical or enzymatic hydrolysis. The system is rel...

We wish to identify genes associated with disease. To do so, we look for novel genes whose expression patterns mimic those of known disease-associated genes, a method we call Guilt-by-Association (GBA). GBA uses a combinatoric measure of association that provides superior results to those from correlation measures used in previous expression analyses. Using GBA, we have examined the expression of 40,000 human genes in 522 cDNA libraries, and have identified several hundred genes associated with known cancer, inflammation, steroid-synthesis, insulin-synthesis, neurotransmitter processing, matrix remodeling and other disease genes. The majority of the genes thus discovered show no significant sequence similarity to known genes, and thus could not have been identified by homology searches. We present here an example of the discovery of five genes associated with schizophrenia and Parkinson's disease. Of the 40,000 most-abundant human genes, these five genes are the most closely lin...

BACKGROUND Recent studies show that the haptoglobin phenotype in individuals with diabetes mellitus is an important factor for predicting the risk of myocardial infarction, cardiovascular death, and stroke. Current methods for haptoglobin phenotyping include PCR and gel electrophoresis. A need exists for a reliable method for high-throughput clinical applications. Mass spectrometry (MS) can in principle provide fast phenotyping because haptoglobin α 1 and α 2, which define the phenotype, have different molecular masses. Because of the complexity of the serum matrix, an efficient and fast enrichment technique is necessary for an MS-based assay. METHODS MALDI plates were functionalized by ambient ion landing of electrosprayed antihaptoglobin antibody. The array was deposited on standard indium tin oxide slides. Fast immunoaffinity enrichment was performed in situ on the plate, which was further analyzed by MALDI-TOF MS. The haptoglobin phenotype was determined from the spectra by embe...

Bionanotechnology aims to impart new properties to materials from unique functionalities present in biomolecules. However, the promise of bionanotechnology has not materialized beyond the biomedical field due in large part to issues of scalability, purity, and cost of manufacturing. In this work we demonstrate an approach to co‐engineer production and system functionality into a single polypeptide. We designed a system to anchor particles onto hair via a multifunctional polypeptide composed of two domains, one with affinity to hair and the other capable of strong interactions with the particle surface. These strong interactions, exemplified by resistance to anionic surfactants, stem from the ability to self‐assemble into higher order structures, which were observed by atomic force microscopy. At the same time, the controlled solubility properties of the particle binding domain permit the scalable production in Escherichia coli via inclusion bodies and cost effective purification. We...

Objective: To collect data on the practices of molecular genetic testing (MGT) laboratories for the development of national and international policies for quality assurance (QA). Methods: A web-based survey of MGT laboratory directors (n = 827; response rate 63%) in 18 countries on 3 continents. QA and reporting indices were developed and calculated for each responding laboratory. Results: Laboratory setting varied among and within countries, as did qualifications of the directors. Respondents in every country indicated that their laboratory receives specimens from outside their national borders (64%, n = 529). Pair-wise comparisons of the QA index revealed a significant association with the director having formal training in molecular genetics (p < 0.005), affiliation with a genetics unit (p = 0.003), accreditation of the laboratory (p < 0.005) and participation in proficiency testing (p < 0.005). Research labs had a lower mean report score compared to all other settings (...

BackgroundTo date, the usage of Galaxy, an open-source bioinformatics platform, has been reported primarily in research. We report 5 years’ experience (2015 to 2020) with Galaxy in our hospital, as part of the “Assistance Publique–Hôpitaux de Paris” (AP-HP), to demonstrate its suitability for high-throughput sequencing (HTS) data analysis in a clinical laboratory setting.MethodsOur Galaxy instance has been running since July 2015 and is used daily to study inherited diseases, cancer, and microbiology. For the molecular diagnosis of hereditary diseases, 6970 patients were analyzed with Galaxy (corresponding to a total of 7029 analyses).ResultsUsing Galaxy, the time to process a batch of 23 samples—equivalent to a targeted DNA sequencing MiSeq run—from raw data to an annotated variant call file was generally less than 2 h for panels between 1 and 500 kb. Over 5 years, we only restarted the server twice for hardware maintenance and did not experience any significant troubles, demonstra...

Aim: To investigate the photodynamic therapeutic potential of ferromagnetic iron oxide nanorods (FIONs), using Trigonella foenum-graecum as a reducing agent, against Leishmania tropica. Materials & methods: FIONs were characterized using ultraviolet visible spectroscopy, x-ray diffraction and scanning electron microscopy. Results: FIONs showed excellent activity against L. tropica promastigotes and amastigotes (IC50 0.036 ± 0.003 and 0.072 ± 0.001 μg/ml, respectively) upon 15 min pre-incubation light-emitting diode light (84 lm/W) exposure, resulting in reactive oxygen species generation and induction of cell death via apoptosis. FIONs were found to be highly biocompatible with human erythrocytes (LD50 779 ± 21 μg/ml) and significantly selective (selectivity index >1000) against murine peritoneal macrophages (CC50 102.7 ± 2.9 μg/ml). Conclusion: Due to their noteworthy in vitro antileishmanial properties, FIONs should be further investigated in an in vivo model of the disease.

Protein A affinity chromatography is a core unit operation in antibody manufacturing. Nevertheless, there is not enough understanding of in‐column antibody adsorption in the Protein A capture step. This work aims to investigate in situ the establishment of an antibody (trastuzumab) layer during Protein A chromatography both in terms of energetic contributions and uptake kinetics. Flow microcalorimetry is employed as a technique with an in situ operating detector, which provides an understanding of the thermodynamics of the adsorption process. In addition, the antibody uptake rate is also investigated in order to establish a correlation between its diffusion on the stationary phase and the associated thermodynamics. Two resins with different particle size, intraparticle porosity, and a Protein A ligand structure are studied: the synthetically engineered B‐domain tetrameric MabSelect SuRe and the synthetically engineered C‐domain hexameric TOYOPEARL AF‐rProtein A HC. The uptake rate f...

Information systems are currently developing very rapidly, and this is inseparable from the role of internet of things (IoT) technology, especially in the world of telemedicine. MitApp is an open-source application that can be used to monitor electrocardiogram (ECG) signals in real-time. The aim of this study is to develop an IoT-based ECG signal monitoring system that utilizes the MitApp application to detect abnormal ECG signals that are characterized by symptoms of cardiac arrhythmias. To process ECG signal data obtained from lead electrode results, the research method utilizes Arduino Uno as a microcontroller. The result is then displayed on the thin film transistor (TFT) layer using the Nextion module. The ESP32 module is used as a Wi-Fi module to send data to the MitApp app on a smartphone. The results showed that the results of the comparison test of ECG signal module data with ECG simulator tools with beats per minute values of 60, 80, 100, 120, and 140 obtained an error rate of <0.05. Based on these results, there is potential to develop this feature and integrate the system with the patient management system to improve the effectiveness of remote monitoring.

Background: Bacteriocins (Bac1, Bac2, and Bac3) from Weissella confusa MBF8-1, weissellicin- MBF, have been reported as potential alternative substances as well as complements to the existing antibiotics against many antimicrobial-resistant pathogens. Previously, the genes encoded in the large plasmid, pWcMBF8-1, and the spermicidal activity of their synthetic peptides, originally discovered Indonesia, have been studied. Three synthetic bacteriocins peptides of this weissellicin-MBF have been reported for their potential activities, i.e. antibacterial and spermicidal. Objective: The aim of this study was to construct the recombinant Bacteriocin (r-Bac) genes, as well as to investigate the gene expressions and their functional analysis. Method: Here, the recombinant Bacteriocin (r-Bac) genes were constructed and the recombinant peptides (r-Bac1, r-Bac2, and r-Bac3) in B. subtilis DB403 cells were produced on a large scale. After purification, using the His-tag affinity column, their ...

Background: Urinary excretion of d-xylose by suckling rats after ingestion of a mixture of 4-, 3-, and 2-galactosylxyloses reflects lactase activity in vivo. We aimed to select the most convenient of these disaccharides for detecting changes of the enzyme activity in vivo and to optimize the method. Methods: 4-, 3-, and 2-Galactosylxyloses were synthesized and purified, then orally administered to suckling rats of different ages. d-Xylose was measured colorimetrically by the phloroglucinol reaction in urine and plasma. Lactase activity was determined in extracts of small intestine mucosa with lactose, galactosylxyloses, and phlorizin as substrates. Results: d-Xylose appeared in the urine in a dose-dependent manner after ingestion of any of the 3 galactosylxylose disaccharides. Correlation between d-xylose elimination and intestinal lactase activity was highest with 4-galactosylxylose (r = 0.97; n = 24), lower with 2-galactosylxylose (r = 0.89; n = 24), and lowest with 3-galactosylxy...

Background: A nosocomial infection, is an infection acquired by a patient during a hospital visit (or among hospital staff) that becomes clinically evident after 48 hours of hospitalization. Objective: To assess the knowledge and practices of health care professionals about nosocomial infections. Methodology: A cross-sectional study with a convenient sample of 250 health care providers was conducted in two tertiary care hospital of Lahore (Sir Ganga Ram and mayo hospital) from November 2015 to January 2016. Out of 250 respondents, 244 replied and participation was voluntary. SPSS version 20 was used for data entry and analysis. Means ± SD were calculated for continuous data and frequency and percentage was used for categorical data. Results: Overall response rate was 97.6%,with majority (89.3%) of the respondents being females. Physicians comprised, 79.50% of the respondents while 20.5% were nurses. Among the physicians more than half were house officers (60.7%).Most of the responde...