Various Types Of Biological Materials Research Articles

Zinc, Copper, and Iron in Selected Skin Diseases.

Trace elements are essential for maintaining the body's homeostasis, and their special role has been demonstrated in skin physiology. Among the most important trace elements are zinc, copper, and iron. A deficiency or excess of trace elements can be associated with an increased risk of skin diseases, so increasing their supplementation or limiting intake can be helpful in dermatological treatment. In addition, determinations of their levels in various types of biological material can be useful as additional tests in dermatological treatment. This paper describes the role of these elements in skin physiology and summarizes data on zinc, copper, and iron in the course of selected, following skin diseases: psoriasis, pemphigus vulgaris, atopic dermatitis, acne vulgaris and seborrheic dermatitis. In addition, this work identifies the potential of trace elements as auxiliary tests in dermatology. According to preliminary studies, abnormal levels of zinc, copper, and iron are observed in many skin diseases and their determinations in serum or hair can be used as auxiliary and prognostic tests in the course of various dermatoses. However, since data for some conditions are conflicting, clearly defining the potential of trace elements as auxiliary tests or elements requiring restriction/supplement requires further research.

Evaluation of engraftment and growth dynamics of orthotopic and heterotopic in vivo models of human breast cancer

Purpose of the study. This work was to assess the engraftment and growth dynamics of breast cancer xenografts during orthotopic and subcutaneous injection using various types of biological material, as well as to develop an adequate model of breast cancer for further research.Materials and methods. We used a disaggregated fragment of a tumor obtained from the patient, a certified breast cancer cell line VT20 – human breast carcinoma; a primary human breast carcinoma cell line. Female immunodeficient mice of the Balb/c Nude line in the amount of 36 animals were used as recipient animals. The subcutaneous and orthotopic models of breast cancer were developed in this project. Tumor growth was observed for 28 days from the moment of injection and tumor nodes were measured 2 times a week until the end of the experiment. Results were assessed using medians and percentiles. The nonparametric Mann-Whitney test was used to assess the significance of differences.Results. The dynamics of the growth of tumor cells when injected into various sites was determined in the process of this work. The most successful in terms of a subcutaneous injection was the injection of tumor cells of the certified VT20 line. By the end of the experiment, the median tumor node of this group was 100.32 mm³. The analysis revealed tumor dynamics with orthotopic injection of tumor material, and the median volume of the tumor node in the group with the passport culture cell VT20 and the primary culture cell reached the same value – 149.22 and 148.25. mm³. It was found that both the cell line and the cell suspension were injected into tumor nodes that reached a significantly larger volume when injected orthotopically.Conclusion. We have obtained a tumor model of breast cancer using various methods of material implantation and with the possibility of further use in testing new pharmacological substances.

Gene-centric coverage of the human liver transcriptome: QPCR, Illumina, and Oxford Nanopore RNA-Seq.

It has been shown that the best coverage of the HepG2 cell line transcriptome encoded by genes of a single chromosome, chromosome 18, is achieved by a combination of two sequencing platforms, Illumina RNA-Seq and Oxford Nanopore Technologies (ONT), using cut-off levels of FPKM > 0 and TPM > 0, respectively. In this study, we investigated the extent to which the combination of these transcriptomic analysis methods makes it possible to achieve a high coverage of the transcriptome encoded by the genes of other human chromosomes. A comparative analysis of transcriptome coverage for various types of biological material was carried out, and the HepG2 cell line transcriptome was compared with the transcriptome of liver tissue cells. In addition, the contribution of variability in the coverage of expressed genes in human transcriptomes to the creation of a draft human transcriptome was evaluated. For human liver tissues, ONT makes an extremely insignificant contribution to the overall coverage of the transcriptome. Thus, to ensure maximum coverage of the liver tissue transcriptome, it is sufficient to apply only one technology: Illumina RNA-Seq (FPKM > 0).

The World of Pseudogenes: New Diagnostic and Therapeutic Targets in Cancers or Still Mystery Molecules?

Pseudogenes were once considered as “junk DNA”, due to loss of their functions as a result of the accumulation of mutations, such as frameshift and presence of premature stop-codons and relocation of genes to inactive heterochromatin regions of the genome. Pseudogenes are divided into two large groups, processed and unprocessed, according to their primary structure and origin. Only 10% of all pseudogenes are transcribed into RNAs and participate in the regulation of parental gene expression at both transcriptional and translational levels through senseRNA (sRNA) and antisense RNA (asRNA). In this review, about 150 pseudogenes in the different types of cancers were analyzed. Part of these pseudogenes seem to be useful in molecular diagnostics and can be detected in various types of biological material including tissue as well as biological fluids (liquid biopsy) using different detection methods. The number of pseudogenes, as well as their function in the human genome, is still unknown. However, thanks to the development of various technologies and bioinformatic tools, it was revealed so far that pseudogenes are involved in the development and progression of certain diseases, especially in cancer.

Fourier Transform Infrared Spectroscopy as a Cancer Screening and Diagnostic Tool: A Review and Prospects.

Infrared spectroscopy has long been used to characterize chemical compounds, but the applicability of this technique to the analysis of biological materials containing highly complex chemical components is arguable. However, recent advances in the development of infrared spectroscopy have significantly enhanced the capacity of this technique in analyzing various types of biological specimens. Consequently, there is an increased number of studies investigating the application of infrared spectroscopy in screening and diagnosis of various diseases. The lack of highly sensitive and specific methods for early detection of cancer has warranted the search for novel approaches. Being more simple, rapid, accurate, inexpensive, non-destructive and suitable for automation compared to existing screening, diagnosis, management and monitoring methods, Fourier transform infrared spectroscopy can potentially improve clinical decision-making and patient outcomes by detecting biochemical changes in cancer patients at the molecular level. Besides the commonly analyzed blood and tissue samples, extracellular vesicle-based method has been gaining popularity as a non-invasive approach. Therefore, infrared spectroscopic analysis of extracellular vesicles could be a useful technique in the future for biomedical applications. In this review, we discuss the potential clinical applications of Fourier transform infrared spectroscopic analysis using various types of biological materials for cancer. Additionally, the rationale and advantages of using extracellular vesicles in the spectroscopic analysis for cancer diagnostics are discussed. Furthermore, we highlight the challenges and future directions of clinical translation of the technique for cancer.

Flotation of Biological Materials

Flotation constitutes a gravity separation process, which originated from the minerals processing field. However, it has, nowadays, found several other applications, as for example in the wastewater treatment field. Concerning the necessary bubble generation method, typically dispersed-air or dissolved-air flotation was mainly used. Various types of biological materials were tested and floated efficiently, such as bacteria, fungi, yeasts, activated sludge, grape stalks, etc. Innovative processes have been studied in our Laboratory, particularly for metal ions removal, involving the initial abstraction of heavy metal ions onto a sorbent (including a biosorbent): in the first, the application of a flotation stage followed for the efficient downstream separation of metal-laden particles. The ability of microorganisms to remove metal ions from dilute aqueous solutions (as most wastewaters are) is a well-known property. The second separation process, also applied effectively, was a new hybrid cell of microfiltration combined with flotation. Sustainability in this field and its significance for the chemical and process industry is commented.

COMPARATIVE STUDY BETWEEN TOPICAL APPLICATIONS OF LIPOSOMALLY ENTRAPPED DNA REPAIR ENZYMES AND THYMIDINE DINUCLEOTIDES AS RADIOPROTECTORS

The delivery of active agents to the skin by liposome carriers is of great interest during the last three decades, based on t heir pote ntial to enclose various types of biological materials and to deliver them to diverse cell types. Purpose: is to compare between topical application of DNA repair enzymes and t hymidine dinucleotide which is a photo protective agent against non - ionizing radi ation through induction of DNA repair. Materials and Methods: thirty six Albino rats were treated by either thymidine dinucleotides or liposomally entrapped DNA repair enzymes topically, 24 hours after they were irradiated with Cobalt 60(60Co) gamma radiat ion with different doses (0.5 and 3Gy), and they were e valuat ed w ith histolopathologically by H&E stain and c omputerized image analyzer using Masson’s trichrome stain. Results: Gamma radiation produced epidermal thinning and dermal inflammatory cells toget her with collagen fragmentation and clumping in a dose - like manner. T hymidine dinucleotide s gave a better response with both (0.5& 3 Gy) than liposomally entrapped DNA repair enzymes , t he epidermis was preserved of with n o inflammatory cells and also it ma intained the normal architecture of collagen bundles. C onclusion : T he effects of gamma radiation on the skin could be minimized by the use of certain dinucleotides .

Liposomal systems as drug delivery vehicles for dermal and transdermal applications

Enhancement strategies are necessary to improve the dermal/transdermal bioavailability of drugs applied to the skin due to its amazing barrier, the stratum corneum. Strategies to overcome this barrier, thus improving drug release to the skin include the use of penetration enhancers, specific delivery systems, supersaturated solutions and physical methods (iontophoresis, electroporation and ultrasound). Delivery of active agents to the skin by liposomal carriers has improved topical therapy in the field of dermatology. The interest in these carriers is based on their potential to enclose various types of biological materials and to deliver them to diverse cell types. Particularly, in recent years liposomes have been shown to be a promising drug-delivery system to the skin. Their use may produce several-fold higher drug concentrations in the epidermis and dermis and lower systemic concentrations when compared to conventional dosage forms. On the other hand, special characteristic vesicles like ethosomes, transfersomes and niosomes may be potential transdermal delivery systems for ionic molecules and polypeptides.

Automated extraction of DNA from reference samples from various types of biological materials on the Qiagen BioRobot EZ1 Workstation

We have validated and implemented a protocol for DNA extraction from various types of biological materials using a Qiagen BioRobot EZ1 Workstation. The sample materials included whole blood, blood from deceased, buccal cells on Omni swabs and FTA Cards, blood on FTA Cards and cotton swabs, and muscle biopsies. The DNA extraction was validated according to EN/ISO 17025 for the STR kits AmpFℓSTR ® Identifiler ® and AmpFℓSTR ® Yfiler ® (Applied Biosystems). Of 298 samples extracted, 11 (4%) did not yield acceptable results. In conclusion, we have demonstrated that extraction of DNA from various types of biological material can be performed quickly and without the use of hazardous chemicals, and that the DNA may be successfully STR typed according to the requirements of forensic genetic investigations accredited according to EN/ISO 17025.

Candida isolates in tertiary hospitals in northeastern Brazil

Candida is an opportunistic pathogen that affects high-risk patients who are either immunocompromised or critically ill and is associated with almost 80% of all nosocomial fungal infections, representing the major cause of fungemia with high mortality rates (40%). Candida albicans is the main cause of candidemia and among the non-albicans species C. parapsilosis, C. glabrata and C. tropicalis are the most frequent agents. The aim of this study was to evaluate the distribution of Candida species in two tertiary hospitals in Recife, Northeastern Brazil. It began by surveying all positive Candida cultures processed by the microbiology laboratory from September 2003 to September 2006. The cultures, originated from various types of biological material (blood, urine, tracheal, catheter and others), were processed by Vitec (®) system (Biomerieux SA, France). A total of 1.279 (hospital A: 837; hospital B: 442) sample isolates were positive for Candida. The most frequent species in both hospitals were: C. albicans (367), C. tropicalis (363), C. parapsilosis (147), C. glabrata (81), C. krusei (30) and C. guillermondii (14). The isolates were obtained from 746 hospitalized patients. A total of 221 positive hemocultures were detected in 166 different patients in both hospitals, and 113 (68.1%) of these patients with positive hemocultures presented Candida in other body sites. This study shows that Candida non-albicans was the main isolated agent and evidences the importante of C. tropicalis in nosocomial fungal infections.

Different types—Different rights

Drawing on a social construction theory of ownership in biological material this paper discusses which differences in biological material might motivate differences in treatment and ownership rights. The analysis covers both the perspective of the person from whom the material originates and that of the potential recipient. Seven components of bundles of rights, drawing on the analytical tradition of Tony Honoré, and their relationship to various types of biological material are investigated. To exemplify these categories the cases of a heart, a kidney, stem cells and hair are used.

High-performance liquid chromatography determination of pipecolic acid after precolumn ninhydrin derivatization using domestic microwave

A novel procedure to specifically quantify low amounts of pipecolic acid and structurally related compounds in several types of biological materials has been characterized. From crude extracts of various types of biological material, the first step was to clear all low-molecular-weight compounds containing primary amino groups by a treatment of nitrous acid. Using a microwave-assisted reaction, the remaining substances containing secondary amino groups were then derivatized with ninhydrin and made soluble in glacial acetic acid. The derivatives produced were resolved by reverse-phase HPLC and detected by spectrophotometry at 570 nm. This procedure allowed more rapid determination of pipecolic acid since microwave heating shortened the time needed for derivatization compared with heating at 95 °C in a water bath. The complete analysis of the chromogens for pipecolic acid and related substances was achieved in 20 min. Under such conditions, the detection threshold for pipecolic acid was about 20 pmol. The suitability of the technique was assessed in various biological matrices known to contain significant amounts of this amino acid. The data obtained are in accordance with those available in the literature. To our knowledge, this is the first method using the ninhydrin reaction in a precolumn, microwave-assisted derivatization procedure for detection and determination of heterocyclic α-amino acids.

Efficient and Reliable PCR-Based Detection of the ABO Blood Group Alleles: Genotyping on Stamps and Other Biological Evidence Samples

PCR-based ABO genotyping was established using restriction enzyme digestion followed by horizontal polyacrylamide gel electrophoresis and silver staining. The method described here is fast, with results obtained within hours, not days, it obviates the need for radioisotopes and can be performed with 1-2 ng of extracted genomic DNA. ABO blood group determination was successful in various types of biological materials of forensic interest such as bloodstains, vaginal swabs, cigarette butts, and hair roots. Moreover, after preincubation in distilled water, DNA (2-8 ng) was extracted from 12 up to 10-years-old stamps and was correctly typed at the ABO locus. The results presented here indicate that the PCR-based ABO genotyping is a fast, sensitive, reliable, and economic method providing blood group determination in DNA from a variety of different types of specimens. It can provide determination from specimens of limited amount and/or with partially degraded DNA as well. Therefore, it is very useful for first-step suspect screening as well as in forensic research for the analysis of biological evidence.

Sex Determination of Forensic Samples by Simultaneous PCR Amplification of α-Satellite DNA from Both the X and Y Chromosomes

Simultaneous amplification of the alphoid repeated sequences clustered in the centromeric regions of both the human X and Y chromosome was performed. Modification and improvement of the polymerase chain reaction conditions resulted in detectable amplification products from less than 1 ng of genomic DNA template. Sex determination was successful in various types of biological materials of forensic interest as bloodstains, vaginal swabs, cigarette butts, bones, and hair roots. The authors suggest that the coamplification of both X- and Y-sequences in a unique reaction mixture is a fast, human specific, sensitive and reliable method providing internal reaction control and sex determination in DNA from a variety of different types of specimens as well as from specimens of limited amount, thus, being very useful in forensic research for the analysis of biological evidence.

The Chemical Oxygen Demand of Waters and Biological Materials from Ponds

Abstract The rate of oxygen consumption by organisms in pond waters, as measured by a dark bottle technique for a 24-hr period, increased with increasing chemical oxygen demand (COD). Rate of oxygen consumption increased with temperature between 15 and 35 C (Q10values were 1.5 to 2.0). At the same level of COD, oxygen consumption was greater in unaltered pond water than in water in which the plankton was heat-killed and oxygen was utilized only by bacteria and other decomposers. COD was positively correlated with increasing concentrations of chlorophyll in pond waters. The amount of COD in soluble organic matter ranged from 20.2 to 95.0% of the total COD. The percentage of the COD in the soluble fraction decreased with increasing quantities of phytoplankton. The average amounts of oxygen (mg oxygen/mg dry wt) required to completely oxidize various types of biological material were; phytoplankton from pure cultures 1.29, macroscopic algae 1.00, higher aquatic plants 0.99, particulate matter from pond water...

Use of calmagite for the determination of traces of magnesium in biological materials

A method has been worked out to determine traces of magnesium in some biological materials. It is based on the use of calmagite as an indicator and EGTA as a masking agent for calcium, which is generally found associated with magnesium. Though the method is liable to interference due to Cu 2+, Zn 2+, and Mn 2+ ions, such limitation may not be of practical significance as these are normally present in traces. The magnesium-dye complex absorbs maximally at 525 mμ. The sensitivity of the method is found to be 0.00123 μg/cm 2. The quantity of sample required for the analysis is of the order 100 mg for body tissues, 50 μl for serum and 10 μl each for red blood cells, urine, and milk. The validity of results has been based on recovery tests and comparison with the available literature values. Further, the results agree when test solutions are prepared in different ways, such as ashing, extraction with TCA or tungstic acid, and simply diluting with water, depending on the nature of sample. In the case of serum and milk samples, the Titan yellow method was used as a reference. The method proves to be simpler and more sensitive than the various known methods and is easily applicable to various types of biological material for routine analysis.

The determination of nano-amounts of iodine in waters, plants, foods, special diets, tissues, and soils

A universal method for the determination of nano-amounts (ng= =10−9 g) of iodine in various types of biological material is described. The recovery of added iodine standard averaged around 95%. The standard deviation for the method is ± 5%.

NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: VIII. DESOXYRIBONUCLEIC ACID PHOSPHORUS AS A MEASURE OF CELL MULTIPLICATION IN REPLICATE CULTURES

A chemical method of estimating the size of cell populations in replicate cultures has been devised for use with certain cell strains from the mouse and rat. The method is based upon the determination of desoxyribonucleic acid phosphorus (DNAP). Although the method may be used as an alternative to the procedure of counting isolated, stained nuclei in a hemocytometer, it actually supplements such measurements and provides useful information on the DNA-content per cell. The method consists of a modified Schmidt and Thannhauser separation of ribonucleic acid (RNA) and desoxyribonucleic acid (DNA) followed by the spectrophotometric estimation of the orthophosphate. The present communication describes the procedures in detail and reports recoveries of RNAP and DNAP from mixtures of highly polymerized nucleic acids and various types of biological material. Good correlation was found between nuclear counts and DNAP values in natural media (containing horse serum and chick embryo extract) and in synthetic (chemically-defined) media. Finally, the efficiency of the dispensing apparatus most frequently employed in preparing replicate cultures from continuously stirred, washed cell suspensions has been determined by measuring the DNAP content of serially dispensed samples.

NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: VIII. DESOXYRIBONUCLEIC ACID PHOSPHORUS AS A MEASURE OF CELL MULTIPLICATION IN REPLICATE CULTURES

A chemical method of estimating the size of cell populations in replicate cultures has been devised for use with certain cell strains from the mouse and rat. The method is based upon the determination of desoxyribonucleic acid phosphorus (DNAP). Although the method may be used as an alternative to the procedure of counting isolated, stained nuclei in a hemocytometer, it actually supplements such measurements and provides useful information on the DNA-content per cell. The method consists of a modified Schmidt and Thannhauser separation of ribonucleic acid (RNA) and desoxyribonucleic acid (DNA) followed by the spectrophotometric estimation of the orthophosphate. The present communication describes the procedures in detail and reports recoveries of RNAP and DNAP from mixtures of highly polymerized nucleic acids and various types of biological material. Good correlation was found between nuclear counts and DNAP values in natural media (containing horse serum and chick embryo extract) and in synthetic (chemically-defined) media. Finally, the efficiency of the dispensing apparatus most frequently employed in preparing replicate cultures from continuously stirred, washed cell suspensions has been determined by measuring the DNAP content of serially dispensed samples.