Dr. Telat Yanik
Department of Aquaculture, Faculty of Fisheries, Atatürk University, Turkey.

ISBN 978-93-91473-45-7 (Print)
ISBN 978-93-91473-46-4 (eBook)
DOI: 10.9734/bpi/rrab/v11

This book covers key areas of biological research. The contributions by the authors include low level laser therapy, Bone histomorphometry, Bone repair, cell-free DNA, necrotic - Piwi interacting, folliculogenesis and biomarkers, treeline ecotone, climate change, snow cover phenology, repeat photography, age structure, hyperglycemia, anti-diabetic, phytochemicals, anti-diabetic potentiality, blood glucose levels analysis, decolorization, enhancers, inhibitors, photoproduction, cancer cell lines, fatty acids, adipocytes, intestinal barrier, fatty acid derivatives, osteopontin, exosome, macrophages,  blood cells count, histopathological changes, sacral hiatus, caudal anaesthesia, anatomical features. This book contains various materials suitable for students, researchers and academicians in the field of biological research.


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Analysing the Effect of Low Level Laser Therapy on Bone Histomorphometry in Rats

Sahar M. Adel, Khaled E. Ayad, Afaf A. Shaheen

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 1-9

The aim of this study was to assess the histological effect of Low Level Laser Therapy (LLLT) (904 nm) on the repair of standardized bone defects on the femur of rats. Sixty male wistar rats were assigned into two equal groups. Group (A: laser group) and group (B: control group). A surgical fracture was done in middle third of femur of all rats. In group (A) a continuous wave 904 nm infrared laser was applied at dose 4 j/cm2 at fracture site immediately post operative for 7 sessions, each session was 5 minutes. The animals were killed by over dose of general anesthesia on the 15th,30th and 45th days after surgery, The specimens were processed and stained with Hematoxylin-eosin (H/E), special stain Masion trichrome and analyzed by light microscopy. The descriptive analysis of histological imaging showed greater degree of new bone formation, osteoblastic surface and collagen fiber in the irradiated group when compared with the control group. Based on the obtained results, this study concluded that LLLT was efficient in promoting bone healing, and increasing new bone formation in the process of surgically fractured femur in animal study.

A morphological comparison of Northern (Salvelinus malma) and Southern (Salvelinus curilus) Dolly Varden chars was performed for 31 plastic and 8 meristic characters. Significant differences were observed in 24 plastic and 7 meristic signs, and in 29 cases the differences were highly significant (P > 0.999). In 11 cases, the differences reach the subspecific level - CD>1.28. Along the length of the upper jaw, there is a chiatus at the limits of variation of the index, and this is already at the level of the species. From the meristic features, it is necessary to distinguish the number of vertebrae – on this basis, the differences between these species are very large: CD=2.33. All this indicates significant differences between these samples and, once again, confirms the species status of southern Dolly Varden - Salvelinus curilus.

Circulating Nucleic Acids: Potential Biomarkers of Male Infertility

Modou M. Mbaye, Bouchra El Khalfi, Achraf Zakaria, Mustafa Zakaria, Noureddine Louanjli, Abdelaziz Soukri

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 17-28

The use of extracellular or circulating nucleic acids (cfs), as a diagnostic or prognostic tool in oncology, has been widely documented. However, their use in gynecology and obstetrics as non-invasive biomarkers in the management of infertility has become recurrent. Free nucleic acids constitute a new source of diagnostic and / or prognostic biomarkers in human pathology. Circulating nucleic acids are made up of: free DNA which can be a long or short strand of DNA resulting from apoptotic or necrotic processes, free RNA containing it: microRNAs (miRNA), which are ribonucleic acids Short single-stranded (RNA) capable of inhibiting the production of proteins from a gene, Piwi interacting RNAs (piRNAs), which are small RNAs expressed in germ cells or even in early embryos, and small RNAs interfering (siRNA), which are small RNAs capable of binding specifically to a messenger RNA sequence and of preventing gene expression by cleaving that RNA. The presence of circulating nucleic acids in many biological fluids such as urine, seminal plasma and serum, the fact that they are easy to detect, the variation in their level depending on the pathophysiological conditions of the body and their involvement in many biological processes such as folliculogenesis, steroidogenesis and spermatogenesis make circulating nucleic acids important and interesting biomarkers for the management of male infertility. They constitute a real complementary aid for the practitioners of medically assisted procreation. Therefore, circulating nucleic acids are a promising avenue in the prevention of implantation failures. In this article we will seek to further assert their importance in the management of male infertility, highlighting their different uses.

In a context of recent climate change, the conversion of a restricted area of treeless alpine/subalpine tundra to mountain birch forest (Betula pubescens ssp. czerepanovii) was studied by repeat photography, demographic analysis in combination with tree growth surveillance in permanent plots. In addition, flora change was recorded within the emerging birch forest stand. The study was initiated in 1980, when a large snow bank still covered the site well into mid-July. Climate warming and associated enhanced snow melt since the early 20th century has made the snow disappear earlier during most summers. In response, a fairly dense population of seed-regenerated low-growing birch saplings gradually emerged. The population stagnated in average height until the early 1980s, when height growth and recruitment accelerated. Thereafter, a dense stand of tree-sized birches emerged. Concurrently, the character of the ground cover transformed from alpine snow bed to subalpine birch forest, as the snow has melted increasingly early, in combination with the presence of a shading tree layer. Earlier snow melt facilitated the establishment of scattered saplings of Pinus sylvestris. Possibly, an analogous course of patchy elevational subalpine forest expansion may be suggested as a reasonable option in a hypothetical case of further climate warming. The establishment of this forest stand may bear some resemblance to the evolution if the first Holocene mountain birch forests.

Investigating the Anti-diabetic Effect of Leucaena leucocephala Linn. (Ipil-Ipil) Seed Extract on Albino Mice

Elchin R. Juanico, Abel Alejandro U. Flores, Jr.

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 45-50

The anti-diabetic effect of Leucaena leucocephala Linn. (Ipil-ipil) seed extract was demonstrated using the Randomized Complete Block Design (RCBD) on albino mice as test animals. The researchers also wanted to identify the phytochemicals in the extract, determine its physical qualities, and compare its effects at three concentration levels with a positive control, a commercially manufactured anti-hyperglycemic drug. The chemical characteristics of the extract revealed the presence of alkaloid, flavonoid, saponin, tannin, and phenolic components, but no anthraquinone or steroid. The extract was orange in colour, had a pleasant flavour, had a pH of 5.44, and was found to be miscible in water and acetone but not chloroform.  The 50% level of extract was found to be the most successful in lowering blood glucose levels (BGL) in the experimental animals, whereas the pure extract (100% concentration) was found to be the least effective. However, there were no significant differences in BGL reductions between the concentration levels and the commercial drug. As a result, Leucaena leucocephala Linn clearly displays anti-diabetic properties comparable to those of the commercial anti-hyperglycemic medication Metformin.

Additional parts of the plant should be evaluated for anti-diabetic potential, taking into account the identification of other secondary metabolites that may be present in the plant components, as well as evaluating its other potential applications.

The present study was aimed to find out the ability of the marine cyanobacterium Oscillatoria boryana BDU 92181 to produce H2O2 and to optimize its photoproduction with a view to harness its potential for decolorization of distillery effluent. The organism decolorized distillery effluent (5% V/V) by about 60% in 30 days. The mechanism of decolorization is postulated to be due to the production of hydrogen peroxide and molecular oxygen released by the cyanobacterium during photosynthesis. The effects of light intensity as well as different enhancers and inhibitors on the production of H2O2 in this organism were studied. Our results showed that, riboflavin and MnSO4 positively influenced H2O2 production. Sodium metabisulphite and potassium cyanide inhibited H2O2 production.

Gastrointestinal epithelium is the unique route for nutrients and for many pharmaceuticals to enter the body. The present study aimed to analyze precisely whether co-culture of two colon cancer cell lines, mucus-producing cells HT29-MTX and enterocyte-like Caco-2 cells, ameliorates differentiation into a functional in vitro intestinal barrier model. Experimental validations were performed in real-time experiments, immunochemistry, and gene expression analyses on Caco-2 versus co-cultures of Caco-2 and HT29-MTX (10%) cells. Partial maintenance of cancer-cell phenotype in differentiated Caco-2 cells was confirmed and fatty acids merged as potential regulators of cancer signaling pathways. HT29-MTX cells induced morphological changes in Caco-2 cells, slightly increased their proliferation rate and profoundly modified gene transcription of phenotype markers, fatty acid receptors, intracellular transporters, and lipid droplet components as well as functional responses to oleic acid. In vitro, enterocyte phenotype was partially rescued by co-culture of cancer cells with goblet cells and completed through oleic acid interaction with signaling pathways dysregulated in cancer cells. Such a reconstructed gastrointestinal epithelium was tested on lipid transport. Adipose tissue function in the regulation of lipemia is highly dependent on intestinal absorption of nutrients. Therefore we developed and validated an in vitro multiculture model allowing to measure intestinal absorption using adipocytes as lipid sensors. Oleic acid (OA) was pre-absorbed onto the reconstructed intestinal barrier. Optimized experimental conditions for co-culture with intestinal barriers were obtained with partially differentiated 3T3L1-MBX adipocytes sensing up to 5 µM OA in solution or 40 µM OA pre-absorbed by Caco2/HT29-MTX intestinal barriers. Metabolism including glycemia and insulinemia greatly influenced the ability to accumulate TG in adipocytes. The present study demonstrates a much better functionality for fatty acid uptake and release in Caco2/HT29-MTX versus Caco-2 intestinal barriers. Taken together these results open new opportunities to study in vitro lipid transfer between intestinal barriers and either adipocytes or hepatocytes.  

Efficient Production of Osteopontin-Encapsulating Exosome by LPS Stimulated THP-1 Macrophages

Gaowa Bai, Takashi Matsuba, Toshiro Niki, Toshio Hattori

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 83-96

Osteopontin (OPN) mediates bone remodeling and tissue debridement. The OPN protein is cleaved, but it is unclear how full-length (FL)-OPN or its cleaved form perform their biological activities in target cells. We, therefore, performed the molecular characterization of OPN in exosomes (Exo). The Exo were isolated from lipopolysaccharide (LPS)-stimulated phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophages. The Exo were also isolated from PMA-differentiated THP-1 macrophages. The Exo were identified using the qNano multiple analyzer (diameter 59–315 nm) and western blotting with a CD9 antibody. LPS-stimulated cells produced more particles than non-stimulated cells. The presence of the FL or the cleaved form of OPN was confirmed using western blot analysis. A mixture of FL and cleaved OPN was also measured using an ELISA system (Ud-OPN) and their presence in the Exo was confirmed. Ud/FL ratios became low after LPS stimulation, indicating the enhanced encapsulation of FL-OPN in the Exo by LPS. These findings suggest that LPS stimulation of human macrophages facilitates the synthesis of FL-OPN, which is cleaved in cells or the Exo after release. These findings indicate that Exo is a suitable vehicle to transfer OPN to the target cells. It is necessary to investigate its role in COVID-19 infection.

Meat Quality and Hematological Indices of Oreochromis niloticus Fish Experimentally Exposed to Escherichia coli Toxins: A Recent Study

Amal M. Yacoub, Sherifa Mostafa M. Sabra, Mona Khaled D. Al-Kourashi

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 97-113

The fish is a good source of proteins for human. Fish of Oreochromis niloticus [1] were treated with different concentrations of Enterotoxigenic Escherichia coli (ETEC) as 103-105, 106 -107 and 109-1010 CFU/ml water. The investigation had conducted to detect experimentally the impact of E. coli toxins on Oreochromis niloticus fish throughout cells blood counts (WBCs, RBCs and Platelets), hemoglobin concentration and histopathological changes of muscles. The values of WBCs, RBCs, Platelets and hemoglobin ranged between (90.70 K/ml -7.10 K/ml), (2.00 m/ml - 0.53 m/ml), (21 K/ml - 3 K/ml) and (8.2 G/dl to 2.50 G/dl) for the hematological indices respectively. The concentration of WBCs, RBCs, Platelets and hemoglobin were lower than control in all the experiment phases. Histopathological changes of O. niloticus treated with E. coli included degeneration, necrosis, atrophy and inflammatory infiltration of WBCs. The results revealed mortality of fish in the highest concentration and high incidence of E. coli toxins in the muscles of O. niloticus induced damage in the muscle tissues and may transfer to the human consumer and cause serious illness. It is recommended to prevent fishing in seas or rivers polluted with sewage or usage of sanitation on farms.

Study on Revisiting Sacral Hiatus

Rajani Singh

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 114-120

Sacral hiatus is caudal opening of sacral canal. Caudal epidural anaesthesia for various therapeutic and diagnostic procedures is administered through sacral hiatus. 25% failure rate of caudal epidural block has been reported in literature. The failure rate of this procedure may be due to variant anatomical features of sacral hiatus. Hence knowing the anatomical variations in shape, size, level of apex and base of sacral hiatus is of paramount importance to clinicians to reduce failure rate of caudal epidural block. The anatomical features of sacral hiatus are of immense value to orthopaedic surgeons, obstretricians, paediatricians, anaesthecians and anatomists. Aim of study is to update and consolidate the information on sacral hiatus as this updated and consolidated information on sacral hiatus in this chapter will serve a ready reference for future researchers and clinicians.

Life as Interplay of Information and Matter

Gerard Battail

Recent Research Advances in Biology Vol. 11, 5 July 2021, Page 121-136

Any sequence can be transformed into an equivalent one by alphabet change and/or encoding. Thus an information cannot be identified to a single sequence, but should be defined as the equivalence class among sequences with respect to such transformations, an abstract entity. It may be represented by its ‘information message’ defined as the shortest binary sequence in this class, whose length quantitatively measures the information. Any sequence must be borne by a physical medium, so information bridges the abstract and the concrete. Perturbations in the physical world result in random symbol errors, while information may instruct the assembly of physical objects by the agency of the semantics it bears. Information theory tells that errorless communication is possible provided a long and redundant enough error-correcting code is used. Conserving genetic information over the ages needs error-correcting codes making the exact regeneration of genomes possible provided it is performed frequently enough. In engineering, such codes are conveniently defined by mathematical constraints which make the symbols mutually dependent, but constraints of any kind, defining ‘soft codes’, also do the job. The better conservation of old parts of the genome implies moreover that the genomic error-correcting codes are made of nested component codes. This scheme accounts for basic life features: need for successive generations, existence of discrete species with hierarchical taxonomy, trend of evolution towards increased complexity, etc. Transcription of the genes and their translation into polypeptidic chains, becoming proteins when properly folded, are controlled by enzymes, which as proteins are are needed for their own synthesis. A feedback loop results, referred to as ‘semantic’ since it implements the genetic code which tells the meaning of the genes. Once closed, it keeps its structure, thereby conserving the genetic code. The onset of a new semantic feedback loop originates an organic code in Barbieri’s meaning, which itself induces a new soft component in the system of genomic nested error-correcting codes by imposing new constraints to the genome. Due to the feedback, constraints incurred by the proteins affect the genome. Besides the synthesis of proteins, the assembly of living structures, as instructed by the genome, involves interwoven semantic feedback loops. Although closed, they do not prevent the genome lengthening, say by horizontal genetic transfer, which increases both the information quantity, hence its semantic specificity, and the redundancy, hence its error-correction ability. Thus, closing semantic feedback loops does not prevent evolution: new structures can be appended with improved resilience to errors. Regeneration failure is very infrequent and results in a widely different genome which originates a new species if the phenotype it specifies withstands the Darwinian selection. A possible scenario for the origin of semantic feedback loops (hence maybe of life) is proposed.