Protargol Research Articles

COMPARATIVE ELECTRON-MICROSCOPIC AND MICROBIOLOGICAL INVESTIGATION OF SILVER PROTEINATE PREPARATIONS

The purpose of this work was a comparative study of the particle sizes of commercial preparations of colloidal silver by the method of electron microscopy, in connection with the characteristics of specific antimicrobial activity of the preparations under the same conditions. Four commercial preparations of colloidal silver were investigated: Vitargol Forte (manufactured by NPC «Elusan»), Protalor (manufactured by OOO «Esko-Farm», Armenia), Sialor (manufactured by PFK «Obnovlenie»). Protargol (prescription drugstore) served as the drug of comparison. Material and methods. The silver particle sizes in the preparations were determined by high-resolution transmission electron microscopy using a JEM 2010 electron microscope (JEOL, Japan) with a resolution of 0.14 nm at an accelerating voltage of 200 kV. The antimicrobial activity of the silver protein preparations was studied by a standard serial dilution method in liquid nutrient media and diffusion method using bacterial cultures of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Results and discussion. Vitargol Forte, manufactured using advanced technology, is the most fine-dispersed and monodisperse preparation. The preparation of standard classical Protargol, prepared in pharmacy conditions, was the most coarsely dispersed and polydispersed. The conclusion. The dependence of antimicrobial activity of the preparation on the size of its silver particles was shown. Of the four investigated preparations of silver proteinate, the smallest particle size was in the Vitargol Fort preparation. The same drug showed the highest bactericidal and bacteriostatic activity.

CLINICAL EFFICACY OF PELARGONIUM SIDOIDES EPS® 7630 IN COMPLEX THERAPY OF CHILDREN WITH CHRONIC ADENOIDITIS, WHO HAS UNDERGONE ADENOIDECTOMY

200 children aged 4 to 12 years diagnosed with grade 3 adenoids, chronic adenoiditis, admitted for routine surgical intervention – adenoidectomy, have been examined and treated in the Pediatric ENT-pathology Department of the Moscow Department of Health L.I. Sverzhevskiy Otorhinolaryngology Healthcare Research Institute. 2 groups of patients were formed depending on the treatment performed in the postoperative period. Group I (active treatment): 100 children who underwent conservative treatment after adenoidectomy, including: tested/investigational drug – Umckalor solution (per os), dosing regimen: children aged from 4 to 6 years – 5–10 drops 3 times a day, aged from 6 to 12 years – 10 to 20 drops 3 times a day; topical decongestants (in the age-related dosage); silver proteinate (nasal drops in age-specific doses); secondand third-generation antihistamines (in the age-specific doses). Group II (control): 100 children, who received conservative therapy after adenoidectomy, including: antibacterial therapy (in the age-specific doses) (third-generation cephalosporins); topical decongestants (in the age-specific doses); silver proteinate (nasal drops in age-specific doses); secondand third-generation antihistamines (in the age-specific doses). In both groups, the course of postoperative therapy amounted to 7 days. The clinical studies have shown that in the early postoperative period the clinical efficacy of Umckalor as part of combined therapy in children, who underwent adenoidectomy, involves reducing the duration of reactive events, accelerating the healing of the postoperative nasopharynx wound. The combined antiviral, antibacterial, immunomodulatory, anti-inflammatory, mucolytic and secretory-motor effect of the drug allows Umckalor to be used as an alternative to antibacterial drugs as part of combined therapy in children who underwent adenoidectomy.

Cytological staining of protozoa: a case study on the impregnation of hypotrichs (Ciliophora: spirotrichea) using laboratory-synthesized protargol

ABSTRACTProtargol (silver proteinate) impregnation is a common method used to identify and characterize ciliated protozoa. Unfortunately, chemical companies have stopped producing the ‘strong’ protargol powder used in this method. Based on an in-house protocol for its synthesis published in 2013, more than 10 batches of protargol powder were produced and subsequently applied in taxonomic studies. During these studies, the protocol for protargol powder synthesis was slightly modified and employed a peptone not originally listed in the 2013 protocol. This modification improved the results of the impregnation protocol. Protargol preparations of hypotrichs were optimized by adjusting the pH during staining rather than during the synthesis. The pH was adjusted to 7.5–7.6, and an acetone developer was used. While the conditions used in this study are not completely comparable to those using the commercially produced protargol, access to this information could help researchers investigate the diversity of ciliates, particularly hypotrichs.

Motor deficits are independent of axonopathy in an Alzheimer's disease mouse model of TgCRND8 mice.

There have been an increasing number of reports of non-cognitive symptoms in Alzheimer's disease (AD). Some symptoms are associated with the loss of motor functions, e.g. gait disturbances, disturbed activity level and balance. Consistent with clinical findings, several AD mouse models harboring amyloid pathology develop motor impairment. Although the factors that contribute to the motor deficits have not yet been determined, it has been suggested that axonopathy is one of the key factors that may contribute to this particular feature of the disease. Our previous study found that TgCRND8 mice exhibited profound motor deficits as early as 3 months old. In this study, we explored the possible factors that may be related to motor deficits in TgCRND8 mice. Results from silver, neurofilament and amyloid precursor protein (APP) staining revealed no axonopathy occurred in the brain and spinal cord of TgCRND8 mice at the age of 3 months. Anterograde labeling of corticospinal tract of spinal cord and electronic microscopy (EM) analysis showed that no axonopathy occurred in TgCRND8 mice at the age of 3 months. According to these results, it could be concluded that no axonal alterations were evident in the TgCRND8 mice when motor deficits was overt. Thus, axonopathy may play a less prominent role in motor deficits in AD. These results suggest that mechanisms by which motor function undergo impairment in AD need to be further studied.

Flagelados de termes mexicanos. I. Descripción de algunas especies de Incisitermes marginipennis (Latreille)

The flagellate fauna of Incisitermes marginipennis (Latreille), is described from specimens collected in Mexico D. F. This fauna includes 16 species, 11 of which were considered as new species. The flagellates were studied in vivo and in permanent smears. For the latter, the flagellates were killed with osmium tetraoxide fumes, and fixed witb several fluids. The descriptions are based principally on slides impregnated with protargol or silver nitrate in gelatine. The authors distinguish between Devescovina and Metadevescovina on the basis of the parabasal body structure. The crista was not observed in three species of Devescovina and seven of Metadevescovina, but in all species the intracitoplasmic portian of recurrent flagellum was considerably thicker than the free portion. Only scattered information by Kirby was available for Mexico, before the publication of this paper.

Therapeutic effect of local treatment of chronic catarrhal rhinitis with 2% aqueous solution of silver proteinate

The authors performed a study of comparability of therapeutic effect in chronic catarrhal rhinitis of 2% silver proteinate aqueous solution (protargol) and a modern antiseptic recommended for external application and for applications on mucous membranes. The therapeutic effect is evaluated based on its antimicrobial activity (in vitro), the dynamics of clinical symptoms over the seven-day observation period, morphological and immunohistochemical study of biopsy specimens of the nasal cavity mucosa. It is shown that the antiseptic agents protargol and octenisept have similar antimicrobial effect on microorganisms, typical of chronic catarrhal rhinitis. When applied topically both 2% protargol and octenisept as diluted 1:6 provide restoration of the epithelial barrier and its protective function, with the reduction of stress of morphofunctional state of epithelium with the tendency to normalization.

Improvement of Silver Impregnation Technique Using in Situ Synthesized Protargol

An improved impregnation method for ciliates has been described with a new formulation for silver proteinate synthesized in situ, that avoids necessary its long-time laboratory synthesis or use of expensive commercial protargol. Compared to conventional techniques, the proposed protocol is more time-saving, reduces the consumption of chemicals and excludes some hazardous ones (e.g. xylene). Structures that are impregnated such as nuclear apparatus, infraciliature, cortical and cytoplasmic microtubules are stained almost identical compared to other protargol methods. Advantages of this method allow us to merge it successfully with ecological quantitative studies of various natural communities of ciliates and provide correct identification of species during such investigations.

Ultrastructure of the ovary and oogenesis in the flatworm Prosthiostomum siphunculus (Polycladida, Cotylea).

Based on light and electron microscopy observations, oogenesis in the cotylean polyclad Prosthiostomum siphunculus was investigated for the first time. The numerous ovarian follicles are dispersed essentially in the dorsal parenchyma. In the follicles, a ventral germinative zone with undifferentiated germs cells of different sizes and a dorsal growth zone with larger growing and abortive oocytes are present. The oogenesis could be subdivided into four stages: (1) Oogonia with a dark nucleus and a dark, ribosome-rich cytoplasm. (2) Early oocyte stage, represented by relatively small cells (10 μm in diameter), a cytoplasm showing some mitochondria and some endoplasmic reticula. (3) Previtellogenic stage, with a decrease of the nucleo-cytoplasmic ratio due to the remarkable increase in ooplasm volume. Immature eggshell globules are observed. (4) Vitellogenic stage, including early vitellogenic ovarian stage, in which a second type of globule (inclusion globule) is formed, and a late vitellogenic uterine stage, in which the inclusion globules are not present anymore. The mature eggshell globules form a peripheral layer under the cell membrane. Eggshell and inclusion globules were analyzed with electron energy loss spectroscopy, electron spectroscopic imaging, protease treatment, and with periodic acid thiocarbohydracide silver proteinate to detect polysaccharides. Chromatoid bodies are present in all four stages. For the first time in a flatworm, we provide evidence that accessory cells, forming a tunica around the ovarian follicles, are epithelial or epithelium-like and likely contribute nutrients for the growth of the oocytes.

COMPARATIVE CHARACTERISTICS OF ANTIBACTERIAL EFFECT OF SILVER AND NANOSILVER IN VITRO

Rationale: The problem of the resistance of microorganisms to many classes of antimicrobial agents becomes increasingly threatening. This promotes the search of new formulations for prevention and treatment of infectious inflammation. Aim: To evaluate antibacterial effects of silver nanoparticle colloid solutions on gram-negative, gram-positive and fungal microflora compared to already known formulations based on silver salts and nitrates of other metals. Materials and methods: The effects of silver nanoparticle colloid solutions (with concentration of nanoparticles of 50 and 100 mg/mL, particle diameter of 15±5 nm) on the microorganism growth were studied in Staphylococcus aureus (# 209P), Escherichia coli (# 26941), Klebsiella pneumoniae (# 43062) and clinical isolates of Candida albicans. For comparison, silver proteinate, nitrofural, and solutions of NaNO₃, Sn(NO₃)₂, Co(NO₃)₂ and Zn(NO₃)₂ at equimolar concentrations to AgNO₃ 1% were used. Results: After the plates with test cultures were treated with silver nanoparticle solutions and with comparator solutions, there was sheer culture growth in the areas of silver nanoparticle application (at both concentrations) and no growth in the areas of the comparator solutions application. Conclusion: The results obtained indicate that silver nanoparticle colloid solutions 50 and 100 mg/mL do not influence the growth of the studied cultures, whereas the comparator solutions exert an advanced antibacterial effect.

Biosynthesis Silver Nanoparticles Using Bacillus Amyloliquefaciens Zxw01 and Research on Synthesis Mechanism

This research reported on synthesis of silver nanoparticles using Bacillus amyloliquefaciens zxw01 culture mixed with silver nitrate. The nanoparticles were characterized by UV-vis spectrum, X-ray diffraction (XRD), Transmission electron microscopy (TEM) and high-resolution transmission electron microscope (HRTEM).In addition, we discussed synthesis mechanism by comparing the protein files of the bacteria before and after mixed with silver nitrate and proteins attached to silver nanoparticles. Our results indicated that silver nanoparticles biosynthesized by Bacillus amyloliquefaciens zxw01 were equally distributed with size between 5 nm to 30 nm and face-centred cubic structure; results of SDS-PAGE suggested that after mixed with silver nitrate, the bacteria differentially expressed and produced a new protein with weight of 33 kDa. Furthermore, analysis of proteins attached to silver nanoparticles indicated that protein with weight of 33 kDa was related to the synthesis of silver nanoparticles.

CHOICE OF SILVER DRUGS IN THERAPY OF RHINOSINUSITIS AT CHILDREN AGE

The article describes major etiological factors and mechanisms of rhinosinusitis, traditional approaches to therapy of rhinosinusitis in children. Special attention is given to mechanisms of intense antiseptic, antifungal and immune modulating action of silver. The article presents Protargol drug that can be prepared immediately before application. Due to the silver proteinate with antiseptic, anti-inflammatory and binding effects Protargol can be used for therapy of acute and chronic rhinosinusitis in children.

Morphology, ciliary pattern and molecular phylogeny of Trachelophyllum brachypharynx Levander, 1894 (Litostomatea, Haptoria, Spathidiida)

We isolated a relatively unknown haptorian ciliate, Trachelophyllum brachypharynx , in brackish water from the mouth of the Taehwa River, South Korea. The morphology of this isolate was studied using in vivo observation and protargol impregnation, and its evolutionary history was revealed by phylogenetic analysis of the 18S rRNA gene. The main features of T. brachypharynx include (i) a very narrowly fusiform and slightly contractile body about 380 × 40 μm in size; (ii) two ellipsoidal macronuclear nodules typically connected by a fine strand; (iii) a single terminal contractile vacuole; (iv) filiform extrusomes that are typically 30 µm long; (v) an average of 24 ciliary rows, with two of them anteriorly differentiated into an isostichad dikinetidal dorsal brush; and (vi) hat-shaped lepidosomes. Based on the 18S rRNA gene phylogeny, T. brachypharynx clustered together with Trachelophyllum sp. within the order Spathidiida. Furthermore, phylogenetic trees and networks indicate some members from the genera Enchelyodon and Spathidium as the nearest relatives of trachelophyllids. Therefore, based on the present molecular and comparative-morphological analyses, we suggested a hypothesis explaining how trachelophyllids may have evolved from a spathidiid-like ancestor via an enchelyodonid-like stage.

Laboratory test system for the evaluation of nanomaterial toxicity on Dunaliella salina microalgae

We are reporting a new diagnostic test system for evaluating nanomaterial toxicity on halophile green microalgae Dunaliella salina based on colorimetric measurements enabling us to estimate cell death and growth-rate inhibition in microalgae suspension for serial dilutions of several toxicants. The principle of the method is the photodegradation of chlorophyll from damaged cells, resulting in the discoloration of the cell sus- pension, which enables us to estimate the microalgae population viability. The cell response was studied for two standard pollutants: formaldehyde and hydrogen peroxide. The toxic effect of free CTAB (cetyltrimetilammo- nium bromide, cationic detergent) water solution was compared with the effect of gold nanoparticles: nanorods, nanostars, and nanospheres, all synthesized in CTAB solutions. It was shown that toxicity is mainly caused by free (nonabsorbed on nanoparticles) surfactant molecules. The toxicity on microalgae D. salina decreases in the following order of silver-containing compounds: silver nitrate > silver proteinate >> colloidal silver in same order.

SPECIES COMPOSITION OF PSAMMOPHILIC CILIATES OF SUMGAIT CASPIAN COAST

Comparative data on the species composition and species saprobic indicators of the psammon ciliates from Sumgait coast of the Caspian Sea are given. Methods. “Alive” samples were collected using a small clean plastic wide mouth bottles. Further processing was carried out under laboratory conditions. Small quantities of soil were examined under a binocular microscope MBS-9 . Ciliates detected microvessel caught and fixed castors for further impregnation kinetoma silver nitrate (Chatton et Lwoff, 1930) or silver proteinate. To determine the keys for ciliates used Foyssner’s major publications (Foissner et al., 1991, 1992, 1999) and “Free-living ciliates Atlas” (Alekperov, 2005). Results. We observed 75 species of ciliates during the studies, which species composition and distribution of the collection points are shown in table 1. Diagrams with average data were made relations groups psammophilous ciliates indicators saprobity different zones for each of the sites investigated Sumgait coast. Main conclusions. Environmental analysis using benthic ciliates indicators saprobity different zones showed that as expected, the industrial zone of Sumgayit coast coast, despite the decline in recent years, the total amount of pollution that is still highly contaminated portion of the coastal zone of the Caspian Sea.

Morphology and molecular phylogeny of Pseudouroleptus jejuensis nov. spec., a new soil ciliate (Ciliophora, Spirotrichea) from South Korea

A new soil ciliate, Pseudouroleptus jejuensis, was discovered from Jeju Island, South Korea and described based on live observation, protargol impregnation, and SSU rRNA gene sequence analyses. Pseudouroleptus jejuensis differs from other congeneric species mainly by number of dorsal kineties (5 vs. 4). Based on our observation of late dividers, we confirm that the dorsal kinety anlage 3 forms 3 kineties (i.e., dorsal kineties 3–5), and the dorsal kinety anlagen 1–3 form 3–5/1–2/0 caudal cirri, respectively. Our gene trees support the assignment of this new species in Pseudouroleptus to full supporting values.

Five New Spathidiids (Ciliophora: Haptoria) from Caribbean Tank Bromeliads

There is a widespread belief that spathidiids have few morphological features. In contrast, we show a rich morphological diversity in five new species discovered in tank bromeliads from the Caribbean, using live observation; protargol impregnation; morphometry; scanning electron microscopy; and resting cyst morphology, demonstrating lepidosomes (organic scales) for the first time in spathidiid haptoria. Arcuospathidium bromelicola nov. spec. is very similar to the previously described A. muscorum but its resting cyst has conspicuous, pillar-shaped lepidosomes on the surface. Protospathidium lepidosomatum nov. spec. is very similar to the previously described P. muscicola but has outstanding, nipple-shaped (vs. conical) lepidosomes on the cyst surface. Spathidium bromeliophilum nov. spec., whose ontogenesis is highly similar to that of S. turgitorum, differs from similar species by the body length: width ratio, the number of ciliary rows, the shape of the oral bulge, and details of the ciliary pattern. Spathidium bromelicola nov. spec. is similar to S. muscicola (extrusomes bluntly fusiform and 4 mu m long vs. rod-shaped and > 15 mu m long) and S. stammeri (resting cyst wall smooth vs. spinous). Spathidium wolfi nov. spec. has an anterior and a posterior contractile vacuole. It differs from the supposed nearest relative, S. faurefremieti, by body size (on average 135 x 25 mu m vs. 240 x 17 mu m), the shape of the macronucleus (moniliform vs. a long, tortuous strand), and the total number of dorsal brush bristles (on average 47 vs. 72). The bent oral bulge of Arcuospathidium bromelicola and Spathidium bromeliophilum as well as the occurrence of lepidosomes on the cyst surface of Arcuospathidium bromelicola and Protospathidium lepidosomatum are discussed.

Iatrogenic Rhinopharyngeal Isolated Argyria Induced by Silver-Containing Nasal Drug

Localized nasal argyria has been reported in 3 published articles as secondary to occupational exposure and involving the anterior part of the nose. No previous cases of such pathology involving the rhinopharynx were described. Here we report the first case of localized argyria of the roof and of the posterior wall of the rhinopharynx secondary to prolonged use of nasal drops containing colloidal silver protein. The recognition of such pathology can be useful to increase the number of conditions that must be considered in the differential diagnosis of rhinopharyngeal mucosa alterations.

Differences in the ultrastructural localization of sulfated glycoconjugates between dentine and bone in the ganoid scales of Polypterus senegalus

ObjectivesSulfated glycoconjugates (S-GCs) are essential components of dentinogenesis and osteogenesis, and are involved in the regulation of the mineralization process. The scales of Polypterus senegalus, which are homologous to teeth and phylogenetically derived from ancestral dermal skeleton, comprise true enamel, dentine, and bone. As part of the phylogenic evolutional studies on mesenchyme-derived hard tissues, we investigated the ultrastructural distribution patterns and histochemical properties of S-GCs in the dentine and bone of scales. MethodsFor detection of S-GCs, a high iron diamine–thiocarbohydrazide–silver proteinate (HID–TCH–SP) staining technique was used. ResultsS-GCs were observed in unmineralized predentine and osteoid, mineralized dentine, osteocyte pericellular matrix, but not in mineralized bone and isopedine. Most S-GCs in the dentine and osteoid were susceptible to testicular hyaluronidase, indicating that they are chondroitin sulfates. ConclusionDistinct differences in the S-GC distribution pattern between the dentine and bone were observed. The S-GC distribution pattern associated with the dentinogenesis of scales is very similar to that of mammalian mantle dentine.

Biochemical and histopathological alterations in kidney of rats chronically exposed to mild silver protein

Chronic administration to Wistar rats of Mild Silver Protein (MSP) to a dose of 222 mg Ag/kg/day in the drinking water for six months showed that the MSP produces argyria, with histopathological lesions in the kidney, the organ where silver accumulates. The histopathology of the kidney showed capillary damage, with alteration of the vascular wall, increased perivascular cellularity and area with focal haemorrhage. The studies in 'open field' were carried out after one, three and six months of treatment and did not show modifications in the pattern of spontaneous locomotor activity. Parameters such as uremia, creatininemia, phosphatemia, alkaline phosphatase, aspartate aminotransferase, alanino–aminotransferase, gamma–glutamyl transpeptidase and lactate dehydrogenase registered in plasma displayed normal values showing normal renal and hepatic functionality. The present results encourage the need for future research to elucidate the molecular pathophysiology and potential functional alterations sustained by the morphological changes found.

Taxonomic Revision and Neotypification of Zosterodasys transversus (Kahl, 1928), with Description of a Mirror-Image Doublet (Ciliophora, Phyllopharyngea, Synhymeniida)

The present work describes the morphology and morphometry of a freshwater ciliate, Zosterodasys transversus, using live observation and protargol impregnation. The population from the River Ipeľ, Slovakia is designated as a neotype because (i) no type material is available from the species described by Kahl (1928); (ii) the neotype is consistent with Z. transversus as described by Kahl (1928); (iii) the neotype is from the same biogeographic region as Kahl’s (1928) populations; and (iv) there are several similar species (e.g., Z. agamalievi) whose identity is threatened by the species to be neotypified. The main features of Z. transversus are: (i) a body size of 120–240 × 50–115 μm in vivo; (ii) a broadly to narrowly obovate or elliptical body; (iii) a spherical to very narrowly ellipsoidal macronucleus with a single globular micronucleus nearby; (iv) several scattered contractile vacuoles; (v) an average of 82 ciliary rows, most of which are interrupted by a synhymenium incompletely encircling the body; and (vi) an average of 14 nematodesmal rods strengthening walls of the cyrtos. In the well-growing neotype population, a single cell of a mirror-image type was found. Most of the structures of this monster specimen are similar to those from ordinary cells, except for the double cytostome and cyrtos.