Tissues Of Vertebrate Animals Research Articles

The Role of Endogenous Metal Nanoparticles in Biological Systems.

The blood and tissues of vertebrate animals and mammals contain small endogenous metal nanoparticles. These nanoparticles were observed to be composed of individual atoms of iron, copper, zinc, silver, gold, platinum, and other metals. Metal nanoparticles can bind proteins and produce proteinaceous particles called proteons. A small fraction of the entire pool of nanoparticles is usually linked with proteins to form proteons. These endogenous metal nanoparticles, along with engineered zinc and copper nanoparticles at subnanomolar levels, were shown to be lethal to cultured cancer cells. These nanoparticles appear to be elemental crystalline metal nanoparticles. It was discovered that zinc nanoparticles produce no odor response but increase the odor reaction if mixed with an odorant. Some other metal nanoparticles, including copper, silver, gold, and platinum nanoparticles, do not affect the responses to odorants. The sources of metal nanoparticles in animal blood and tissues may include dietary plants and gut microorganisms. The solid physiological and biochemical properties of metal nanoparticles reflect their importance in cell homeostasis and disease.

Myiasis in a the Tracheotomy Wound: A Case Report

“Myiasis” is a parasitic infestation of live human or vertebrate animal tissues or cavities caused by dipterous larvae (maggots) which feed on the host’s dead or living tissue, liquid body substances or ingested food. The case is reported, probably the first in Bangladesh, tracheostomy myiasis. It was caused by infestation with larvae. The patient had undergone tracheostomy 3 years earlier and was a case of ca-larynx. The condition was treated by applying halothane to the tracheostomy wound, which caused spontaneous exit of approximately 30 larvae, easily removed with forceps. Predisposing factors could be: 1. Immuno-compromization due to post-irradiation state of patient; 2. Poor hygiene of tracheostomy tube; 3. Bad smell of wound, which attracts flies; 4. Living in a rural area deprived from necessary health care. Although this is not a lethal disorder, knowledge of the disease is necessary from the preventive, diagnostic and curative standpoint. It is important to proceed with identification of the larvae, distinguishing them from other types of myiasis involving different therapeutic implications.
 Bangladesh J Otorhinolaryngol; October 2016; 22(2): 119-121

Regulation ranges and patterns of adaptation to hyponatremia by cells of various organs and tissues of vertebrate animals.

The effect of hyponatremia on the body is studied on model objects. The related question concerns the degree of compliance between manifestations of hyponatremia and protective mechanisms in humans and other species of vertebrates. To identify the regulation ranges and patterns of adaptation to hyponatremia by cells of various organs and tissues of vertebrate animals. To assess the regulation ranges and patterns of adaptation to hyponatremia, a comparative analysis has been applied to the data obtained from humans, mammals and freshwater fish. The physiological content of sodium in the blood plasma in humans is regulated and maintained within a narrow value range which is similar to that occurring in a number of other vertebrate species. The counteraction to hyponatremia is performed by means of accelerating the transport of sodium, potassium, chloride and organic osmolytes from the cells into the internal environment. The data regarding mammals are fragmented and reflect the manifestation of protective mechanisms taking place during the initial period of hyponatremia. The method tested on freshwater fish allows for studying patterned changes in inorganic ions and content of organic osmolytes in the internal environment and cells of various organs and tissues of the body from the start of developing hyponatremia till the completion of the recovery process in vivo (Fig. 2, Ref. 72).

Myiasis by <i>Cochliomyia hominivorax</i> (Coquerel, 1858): A Neglected Zoonosis in Brazil

Myiasis is the lesion resulting from the parasitism of diptera larval form in the living tissues of vertebrate animals. These are bloody conditions, causing severe damage to the welfare and the health of humans and animals. In Brazil, Cochliomyia hominivorax is the main responsible species for causing myiasis in humans and animals. The conventional treatment of these lesions in animals massively uses chemosynthetic products. The irrational use of these molecules has led to environmental degradation and has affected human health. The standard treatment of human myiasis is based only on larvae removal, surgically or not, supported by the use of antiparasitic drugs. Human myiasis is an important zoonosis, given its close relation with animal myiasis. However, this zoonosis has currently been neglected in Brazil and other developing countries in America. The One Health approach makes it possible to realize that the occurrence of myiasis in humans is directly related to the maintenance of stocks of this diptera in nature. Recognizing the direct relation that domestic and wild animals have as reservoirs in the human myiasis cycle is essential, in order to formulate strategies to control this ancient and important disease that still affects the population in Brazil.

Life-threatening endobronchial myiasis

Dear Editor, A 47-year-old homeless man was admitted to our intensive care unit (ICU) for septic shock as a complication of an infected diabetic foot ulcer. His previous medical history included a type 2 diabetes mellitus treated with metformine and gliclazide. The patient was found comatose and covered with flies in a shelter on the shore of a river. The prehospital emergency care personnel performed bag-valve mask ventilation with 100% oxygen and rapid sequence induction (intravenous etomidate 20 mg and succinylcholine 100 mg), which enabled successful tracheal intubation by a physician upon the first attempt. The patient was then mechanically ventilated and transferred to our ICU. Upon admission, the patient had a Glasgow coma scale score of 3, blood pressure of 82/45 mmHg, body temperature of 33.7 C, heart rate at 75/min, and arterial oxygen saturation of 90% with an inspired oxygen fraction (FiO2) of 1. A large necrotic wound on the right foot, in which numerous live larvae were found, was noted. Laboratory tests showed lactic acidosis (pH 6.96, blood lactates [15 mmol/L) and acute renal failure (blood urea 42 mmol/L, creatinine 465 lmol/L); the hemoglobin level was 8.3 g/dL and the white blood cell count was 16,300/mm. No abnormalites were found on the chest X-ray. Arterial blood gases obtained after hemodynamic stabilization was achieved (i.e., 1 h after ICU admission) showed normal oxygenation (PaO2/FiO2 ratio 523 mmHg at FiO2 1). Toxicological screening tests were negative. The conclusion was drawn that the comatose state was due to a profound hemodynamic impairment with severe lactic acidosis. Antibiotic therapy and surgical debridement of the wound were performed. Optic microscopy examination identified the collected larvae as Lucilia sericata. Both blood cultures and wound swabs grew methicillin-sensitive Staphylococcus aureus. On the second day, larvae were seen leaking from both of his nostrils, and nasal cavity infestation was confirmed by endoscopy (Fig. 1). A cranial computed tomography (CT) scan revealed there was neither brain nor sinus involvement. Management consisted of mechanical removal of the larvae and a single dose of ivermectine. On the third day of mechanical ventilation, a ventilator-generated pressure alarm revealed a sudden increase in peak airway pressures that partially impeded tidal volume delivery to the patient. Tracheal suctioning was performed immediately and allowed for the extraction of numerous larvae obstructing the endotracheal tube. Flexible bronchoscopy showed the presence of approximately 30 larvae in the left bronchial tree that were then extracted (Fig. 1). Five larvae were removed during another bronchoscopy the following day. Protective measures, including insecticide spray and patient isolation, were implemented to avoid nosocomial transmission. The patient was kept mechanically ventilated in the FiO2 0.3 state because of protracted coma. He finally recovered and was successfully extubated on day 8 and discharged from the ICU on day 11. Myiasis is a parasitic infestation of live human or vertebrate animal tissues or cavities caused by dipterous larvae (maggots). L. sericata, known as the green bottle blowfly, is a facultative ectoparasite. In humans, L. sericata has been involved in nasal [1], wound [2], gastric, tracheostomy [3], and nosocomial [4, 5] infestations. In this case, predisposing factors for myiasis included a necrotizing wound and a protracted coma with lack of the cough reflex. Whether upper airway manipulations (i.e., bag-valve mask ventilation and laryngoscopy) during endotracheal intubation might have favored the

The basal calcium level in fibers of the rat soleus muscle under gravitational unloading: The mechanisms of its increase and the role in calpain activation

The first data on increase in the basal level of cal� cium in immobilized muscles appeared in 1970 (3). Nasledov and coworkers hypothesized that calcium ions accumulated under the conditions of gravita� tional unloading (1). In that study, the authors used the model of antiorthostatic suspension. Using an isolated rat m. soleus, the mechanical response has been dem� onstrated to attenuate rapidly after a series of tetanic contractions in calciumfree Ringer's solution. How� ever, after 14�day unloading in the standard model of antiorthostatic suspension, there was no attenuation in tests performed on muscles of the suspended ani� mals. When, in the second experimental series, Vera� pamil (a calcium channel blocker) was added to the calciumfree medium, attenuation of the mechanical response was observed in the suspended animals. It was suggested that an increased resistance to muscle fatigue in calciumfree Ringer's solution was a result of intracellular accumulation of calcium ions, which freely came out of the muscle through the channels, and Verapamil inhibited this process. Using the calcium probe Fluo�3, a significant increase in the concentration of calcium ions of the mouse m. soleus was later demonstrated as soon as the second day of suspension (8). An excessive amount of calcium ions might occur under microgravity in the myoplasm of the muscle fiber either from the extracellular space through exter� nal calcium channels or from the sarcoplasm reticu� lum because of "leakage" of Ryanodine channels. There is evidence suggesting that Ryanodine channels are involved in increasing the intracellular calcium (12). In addition, a higher expression of dihydropyri� dine receptors was already observed upon shortterm gravitational unloading (9). It was subsequently dem� onstrated that the use of Nifedipine, a selective blocker of calcium channels, against the background of 14�day functional unloading led to a significant diminishing the level of calcium ions in the myoplasm of m. soleus fibers, and it largely prevented the trans� formation of the myosin phenotype (6, 12). At the same time, peroral Nifedipine administration may ini� tiate multiple systemic effects. Because of this, only a decrease in the basal level of calcium ions in the mus� cle fiber in response to a direct Nifedipine action against the background of gravitational unloading can serve as valid evidence of the involvement of slow Ltype calcium channels into accumulation of these ions in fibers of postural muscles. An increase in the calcium ion concentration in the myoplasm of muscular fibers at rest may stimulate the activity of calpains, calciumdependent proteases. Both µ� and mcalpains, which are activated by microand millimolar calcium concentrations, respectively, are present in all tissues of vertebrate ani� mals. These enzymes play the key role in the degrada� tion of a series of cytoskeletal proteins (2). The calpain activity has been demonstrated to grow up signifi� cantly even upon a shortterm gravitational unloading (4, 5), and this was accompanied by a significant decrease in the concentration of desmin, one of the key cytoskeletal muscle proteins. At the same time, the importance for increasing the calpainstimulating activity of calcium ions in the muscle fibers during gravitational unloading, may be understood after application of calcium chelating agents with an intra� cellular activity under these conditions.

Myiasis in the uterine cavity of an elderly woman with a complete uterine prolapse

Myiasis is the infestation of the tissues of vertebrate animals by the larvae of flies (maggots). As the condition generally occurs in exposed areas of the body, genital myiasis is a rare condition, and myiasis of the uterine cavity is even rarer, only one case having been previously described in the literature. The case reported here describes myiasis of the uterine cavity and ulcerated vaginal mucosa of an 89-year-old woman with a severe (third-degree) genital prolapse. This case describes an extremely rare situation, and emphasizes the need for good hygiene and surgical correction in patients with a genital prolapse.

Species identification and authentication of tissues of animal origin using mitochondrial and nuclear markers

We evaluated and compared the utility of mitochondrial markers viz. 16S rDNA and NADH dehydrogenase subunit 4 ( ND4) and a nuclear marker viz. the actin gene to identify the specimens of animal origin for forensic identification, food regulatory control and to prevent illegal trading, poaching and conservation of endangered species. We also tested PCR fingerprinting methods like RAPD and actin barcoding to generate species-specific “fingerprints”. Our results suggested that mitochondrial markers are more efficient than nuclear markers for the purpose of species identification and authentication. Among PCR fingerprinting approaches, RAPD was proved to be more discriminatory, accurate and efficient than actin fingerprinting. Considering the present scenario in trading of vertebrate animal tissues like buffalo, cow, pig, goat, chicken, frogs, fishes and snakes etc., mitogenomics based technology proved to be efficient and reliable in resolving problems like meat adulteration and smuggling across countries.

Molecular cloning and expression of the nag1 gene (N-acetyl-beta-D-glucosaminidase-encoding gene) from Trichoderma harzianum P1.

A 72-kDa N-acetyl-beta-D-glucosaminidase was purified from the mycoparasitic fungus Trichoderma harzianum P1; antibodies were raised against it, and aa-sequences were obtained. The antibody reacted with a single 72-kDa protein band in culture filtrates of T. harzianum grown on chitin, and was subsequently used to clone the corresponding nag1 gene from a lambdagt11 cDNA expression library. It was interrupted by two short introns and encoded a protein of 580 amino acids. The deduced protein sequence contained aa-sequence areas of high similarity to N-acetyl-glucosaminidases from other eukaryotes such as Candida albicans, and invertebrate and vertebrate animal tissues. The highest similarity was observed with the corresponding gene from the silkworm. The aa-sequence of a tryptic fragment of purified N-acetyl-beta-D-glucosaminidase from T. harzianum corresponded to a deduced aa sequence from a portion of the cloned gene, thus verifying that the protein is encoded by nag 1. Southern analysis showed that nag 1 is present as a single-copy gene in T. harzianum. Expression of nag1-mRNA was strongly induced upon growth on chitin, N-acetyl-glucosamine and the cell walls of Botrytis cinerea used as a carbon source. The appearance of the corresponding N-acetyl-beta-D-glucosaminidase protein, as determined by Western analysis, paralleled the pattern of nag 1 expression, thereby suggesting that its formation is regulated at the level of transcription.

Novel mechanism of intracellular calcium release in pituitary cells.

In sea urchin eggs an enzymatic metabolite of beta-NAD+, called cyclic ADP-ribose (cADPR), is as potent and powerful a releaser of sequestered intracellular Ca2+ as is inositol 1,4,5-trisphosphate (IP3). The enzyme that synthesizes cADPR is present in several vertebrate animal tissues, but the Ca(2+)-releasing activity of cADPR has not been described in mammalian cells. We report here that incubation of beta-NAD+ with cell-free extracts of several rat tissues (including pituitary gland) generates a product which releases intracellular Ca2+ stores in permeabilized rat pituitary GH4C1 cells. This product has the biological characteristics of cADPR (it acts after depletion of the IP3 stores and after blockade of the IP3 receptor by heparin). The response is mimicked, in a concentration-dependent manner, by authentic cADPR and is desensitized by prior incubation with cADPR. We conclude that cADPR is not only synthesized by certain mammalian cells but also acts in such cells to release compartmentalized intracellular Ca2+ by a mechanism that differs from that used by IP3. Therefore, cADPR may serve, in addition to IP3, as a second messenger for intracellular Ca2+ mobilization in mammalian cells.

Platelet activating factor‐acetylhydrolase in insects: Identification and partial characterization of a 1‐alkyl‐2‐acetyl‐sn‐glycero‐3‐phosphocholine acetylhydrolase in a cell‐free system of Heliothis

AbstractA specific acetylhydrolase that inactivates platelet activating factor (PAF; 1‐alkyl‐2‐acetyl‐sn‐glycero‐3‐phosphocholine), a potent cellular mediator in mammalian cells, by removal of the sn‐2 acetyl moiety, has been found in the cytosolic fraction of several postembryonic developmental stages and specific tissues of the corn earworm, Heliothis zea (Boddie). Effects of magnesium, calcium, EGTA, deoxycholate, dithiothreitol, diisopropylfluorophosphate, egg phosphatidylcholine, and an acylacetyl‐glycerophosphocholine show that hydrolysis of the acetate moiety is due to a specific acetylhydrolase for PAF. The activity does not appear to be due to a typical cellular phospholipase A2 that utilizes phospholipid substrates with a long‐chain acyl group at position sn‐2 of glycerol. Specific activities and properties of the acetylhydrolase from this insect match closely with those described from tissues of vertebrate animals.

An investigation of the molecular components of desmosomes in epithelial cells of five vertebrates

We have shown previously, by fluorescent antibody staining, that desmosomal antigens are widely distributed in the tissues of vertebrate animals. Furthermore, we have demonstrated mutual desmosome formation between cells derived from man, cow, dog, chicken and frog. In this paper we have studied the components of desmosomes in a tissue or a cell line from each of these animals by immunoblotting with antibodies raised against the desmosomal components isolated from bovine nasal epithelium. Blotting was carried out on bovine nasal epithelial desmosomal cores, desmosome-enriched fractions derived from chicken and frog epidermis, nuclear matrix-intermediate filament scaffolds derived from Madin-Darby bovine and canine cells (MDBK and MDCK), and unextracted cultured human foreskin keratinocytes. The results show that desmosomes from all these sources contain high molecular weight proteins (desmoplakins) of similar or identical molecular weights (250 000 and 215 000). Antibodies against the two lower molecular weight desmosomal proteins (83 000 and 75 000) always recognized one or two bands in very similar molecular weight regions of the gels. The desmosomal glycoproteins were found to be much more variable than the proteins: they vary between sources in molecular weight, heterogeneity and antibody cross-reactivity. For instance, antibody specific for a group of glycoprotein bands of 175 000, 169 000 and 164 000 (Mr) in bovine nasal epithelium recognizes three bands of 245 000, 230 000 and 210 000 in MDCK cells but only a single band of 190 000 in keratinocytes. In mammals, the 175 000-164 000 glycoproteins and the desmosomal adhesion molecules, the desmocollins (Mr 130 000 and 115 000 in cow's nose), are immunologically distinct. In chicken and frog, however, there are glycoproteins that react with both anti-175 000-164 000 and anti-desmocollin antibodies, but there are also distinct desmocollin bands. The significance of these results is discussed in relation to conservation of desmosomal components and adhesion mechanisms. It is suggested that adhesion may be performed by a well-conserved protein domain and that the variation between desmosomal glycoproteins from different sources may be due to differences in their carbohydrate composition.

A study of tissue water metabolism in living organisms by infrared spectroscopy

Infrared absorption spectra of vertebrate animal tissues have been studied in the liquid water band range. A spectroscopic method for determining the water metabolism rate in living organisms has been developed. The technique involves the comparison of light and heavy water bands.