Antiviral and Antibacterial Cold Spray Coating Application on Rubber Substrate, Disruption in Disease Transmission Chain.

The objective of this study was to prepare a copper-coated rubber surface using cold spray technology with improved virucidal and antimicrobial properties to fight against highly transmissible viruses and bacteria. A successful cold spray coating was produced using irregular-shaped pure Cu powder on an escalator handrail rubber. The powder particles and the deposited coatings (single and double pass) were characterized in terms of particle morphology and size distribution, coating surface and coat/substrate cross-section properties. The bonding between powder and rubber surfaces was purely mechanical interlocking. The Cu powder penetration depth within the rubber surface increases with a number of depositions pass. The virucidal properties of the coated surface were tested utilizing surrogates for SARS-CoV-2: HCoV-229E, a seasonal human coronavirus, and baculovirus, a high-titer enveloped insect cell virus. A double-pass coated surface showed significant baculovirus inactivation relative to a bare rubber control surface after 2-h (approximately 1.7-log) and 4-h (approximately 6.2-log), while a 4-h exposure reduced HCoV-229E titer to below the limit of detection. A similar microbial test was performed using E. coli, showing a 4-log microbial reduction after 2-h exposure relative to the bare rubber. These promising results open a new application for cold spray in the health sector. Supplementary Information: The online version contains supplementary material available at 10.1007/s11666-023-01553-x.

Permeability of the ovarian follicle of Aedes aegypti mosquitoes.

The passage of tracers of various molecular weights into resting and vitellogenic ovarian follicles of Aedes aegypti mosquitoes was studied ultrastructurally. The outermost layer of the follicular sheath (the basement lamina) is a coarse mechanical filter. It is freely permeable to particles with molecular weights ranging from 12,000 to 500,000 (i.e. cytochrome c, peroxidase, hemoglobin, catalase, ferritin, immunoglobulin (IgG)-peroxidase, iron dextran and Thorotrast) that have dimensions less than 110 A. Molecules as large as carbon (300-500 A) are totally excluded. Whereas proteins and polysaccharide tracers permeate the basement lamina with apparent ease, certain inert particles (e.g. Thorotrast, Fellows-Testager Div., Fellows Mfg. Co., Inc., Detroit, Mich.) penetrate more slowly. With respect to the tracers tested, resting follicles are as permeable as vitellogenic follicles. The follicle epithelium of resting or vitellogenic follicles is penetrated by narrow intercellular channels. Our observations suggest that these spaces are lined with mucopolysaccharide material. After permeating the basement lamina, exogenous tracers fill these channels, while the bulk of material accumulates in the perioocytic space. Within 3 hr after imbibing blood, the pinocytotic mechanism of the oocyte is greatly augmented. Pinocytosis is not selective with regard to material in the perioocytic space, since double tracer studies show that exogenous compounds are not separated, but are incorporated into the same pinocytotic vesicle. During later stages of vitellogenesis, 36-48 hr after the blood-meal, the pinocytotic mechanism of the oocyte is diminished. Simultaneously, the intercellular channels become occluded by desmosomes, and the vitelline membrane plaques separate the oocyte and follicle epithelium.

The localization of glycogen in the spermatozoa of various invertebrate and vertebrate species.

With the periodic acid-thiosemicarbazide-silver proteinate procedure for the detection of polysaccharides in thin sections, glycogen is localized in the cavities of centrioles and basal bodies, within the axoneme (and surrounding it), in mitochondria, and in the "packing" cytoplasm of the middle piece of spermatozoa of several invertebrate and vertebrate species. The cytochemical localization of glycogen is verified by extraction with alpha-amylase solution. These findings establish the existence of stored glycogen in sperm. The polysaccharide presumably serves as an endogenous source of energy in the absence of extracellular metabolites, under either aerobic or anaerobic conditions. Other hypotheses on the physiological significance of intracellular glycogen stores in sperm are discussed. Sperm that store glycogen contain some enzymes of glycogen metabolism. In the presence of glucose-1-phosphate, ATP, and Mg(++) ions, an amylophosphorylase catalyzes the in vivo synthesis of glycogen. The newly formed product resembles gamma-particles, and is digestible with alpha-amylase.

Induction of endometrial peroxidase synthesis and secretion by estrogen and estrogen antagonist.

Synthesis of peroxidase was induced in the uterine epithelium of immature rats by multiple doses over a 24-96-h period of either 17 beta-estradiol, the estrogen-antagonist Parke-Davis CI-628, or a combination of estradiol plus antagonist. Endogenous peroxidase activity first appeared in the cisternae of the rough endoplasmic reticulum of surface epithelial and glandular cells within 24-48 after the initial injection. Uterine peroxidase activity was also visible in the cisternae of the Golgi apparatus, in Golgi-derived secretory granules, and within the uterine and glandular lumen. Some cells of the epithelium produced little or no peroxidase, even after 96 h. Whereas the antagonist appeared to induce synthesis and secretion of peroxidase, neither the antagonist alone nor the combined treatment (estradiol plus antagonist) reproduced the estradiol-mediated growth in organ size and increased lumen diameter.

Collagenous bone matrix-induced endochondral ossification hemopoiesis.

Transplantation of collagenous matrix from the rat diaphyseal bone to subcutaneous sites resulted in new bone formation by an endochondral sequence. Functional bone marrow develops within the newly formed ossicle. On day 1, the implanted matrix was a discrete conglomerate with fibrin clot and polymorphonuclear leukocytes. By day 3, the leukocytes disappeared, and this event was followed by migration and close apposition of fibroblast cell surface to the collagenous matrix. This initial matrix-membrane interaction culminated in differentiation of fibroblasts to chondroblasts and osteoblasts. The calcification of the hypertrophied chondrocytes and new bone formation were correlated with increased alkaline phosphatase activity and 45Ca incorporation. The ingrowth of capillaries on day 9 resulted in chondrolysis and osteogenesis. Further remodelling of bony trabeculae by osteoclasts resulted in an ossicle of cancellous bone. This was followed by emergence of extravascular islands of hemocytoblasts and their differentiation into functional bone marrow with erythropoietic and granulopoietic elements and megakaryocytes in the ossicle. The onset and maintenance of erythropoiesis in the induced bone marrow were monitored by 59Fe incorporation into protein-bound heme. These findings imply a role for extracellular collagenous matrix in cell differentiation.

Cytodifferentiation during spermiogenesis in Lumbricus terrestris.

The structural changes during spermiogenesis were studied on developing spermatids in seminal vesicles and receptacles of Lumbricus terrestris fixed in glutaraldehyde-osmium tetroxide and embedded in Epon-Araldite. The centriole plays a prominent role in the morphogenesis and organization of the microtubules of the manchette and flagellum. Microtubules arising from the centriole extend anteriorly to encase the developing middle piece, the nucleus, and the acrosome. The manchette not only provides a supporting framework for the cell during elongation, but also may provide the motive force for the elimination of both nucleoplasm and cytoplasm. The manchette participates in segregation and elimination of the nuclear vesicle that contains the nonchromatin nucleoplasm. Compartmentalization and conservation may also be a function of the manchette since those elements which remain within the framework of microtubules are retained, while all the cytoplasm outside the manchette is discarded. At maturation, the endoplasmic reticulum plays a key role in dismantling the manchette and reducing the cytoplasm external to it. During the early stages of middle-piece formation, six ovoid mitochondria aggregate at the posterior pole of the spermatid nucleus. Concurrent with manchette formation, the mitochondria are compressed laterally into elongate wedge-shaped components, and their outer limiting membranes fuse to form an hexagonal framework that surrounds the dense intramitochondrial matrices. Dense glycogen granules are arranged linearly between the peripheral flagellar tubules and the outer membrane of the mature sperm tail.

Effects of acriflavine on the mitochondria and kinetoplast of Crithidia fasciculata. Correlation of fine structure changes with decreased mitochondrial enzyme activity.

The effects of acriflavine on the fine structure and function of the mitochondria and the kinetoplast in Crithidia fasciculata have been investigated. A mitochondrial fraction was prepared by differential centrifugation of cells broken by grinding with neutral alumina. Isolated mitochondria or intact cells revealed by spectrophotometric measurements the presence of cytochromes a + a(3), b, c(555) and o. After cells were grown in acriflavine for 3-4 days, the fine structure of the mitochondria and their cytochrome content were affected. Cells grown in 5.0 microM acriflavine had a threefold decrease in cytochrome a + a(3) and decreased respiratory activity. The mitochondrial preparation from these cells had a fivefold decrease in cytochrome a + a(3) and a less but significant decrease of other cytochromes present. There was also a decrease in the mitochondrial enzyme activities of NADH, succinic and L-alpha-glycerophosphate oxidases, and succinic and L-alpha-glycerophosphate dehydrogenases. Dyskinetoplastic cells could be demonstrated after growth in 1.0 microM acriflavine. At 5 microM, 80-90% of the cells were dyskinetoplastic. The kinetoplastic DNA was condensed, nonfibrillar, and did not incorporate thymidine-(3)H. The mitochondria in these cells had few cristae and were shorter and more swollen than the controls. Acriflavine may induce the fine structure effects we have observed and may affect the formation of the mitochondria in C. fasciculata.

An ultracytochemical study of the respiratory potency, integrity, and fate of the sea urchin sperm mitochondria during early embryogenesis.

Cytochrome oxidase activity via cytochrome c, as demonstrated by the diaminobenzidine procedure, has been employed in this electron microscope cytochemical study to determine the respiratory potency, integrity and fate of the Arbacia sperm mitochondrion at fertilization and during early embryogenesis. The sperm mitochondrion remained intact and was intensely positive for cytochrome oxidase activity both during and after penetration into the egg. The mitochondrion remained highly reactive throughout zygote formation, up to the eight-cell stage. The sperm mitochondrion formed many projections and buds in the cytoplasm of immature oocytes, monospermic and polyspermic eggs, and in blastomeres. At all stages of early embryogenesis, close juxtaposition and structural contact were observed between the highly reactive sperm mitochondrion and the less reactive egg mitochondria. The results suggest that following fertilization the mitochondrion of the sea urchin spermatozoon retains some degree of metabolic autonomy within the ooplasm. The structural integrity of the paternal mitochondrion is maintained along with a functional respiratory enzyme system (cytochrome c-a3). The hypothesis that the fertilizing sperm mitochondrion may have some relevance to sea urchin development is discussed.

Modulation of uterine morphology and growth by estradiol-17beta and an estrogen antagonist.

The estrogen antagonist C1628 maintains sustained hypertrophy of the uterine epithelium and the synthesis of many proteins including peroxidase. C1628 is a progestogen, inducing secretion of the protein by surface epithelial and glandular cells. C1628 is a connective tissue mitogen, inducing DNA synthesis in fibroblasts and the endothelium. C1628 and estrogen share these properties mentioned above. Estrogen, however, induced moderate growth of the mucosa within a 24-h period and massive hyperplasia of the mucosa within a 24-h period thereafter. C1628 alone, or in combination with estradiol, does not have mitogenic effect on the mucosa, and in fact blocks the mitotic response normally induced by estrogen alone.

Endogenous peroxidase: specific marker enzyme for tissues displaying growth dependency on estrogen.

Data derived from a correlated morphological and biochemical study suggest the following: (a) estradiol-17beta, diethylstilbestrol, the estrogen antagonists nafoxidine (Upjohn 11,000), and Parke Davis C1628 induce synthesis of an endogenous peroxidase in the epithelium of target tissues like the vagina, the cervix, the uterus, and in the acinar cells of the estrogen-dependent rat mammary tumor; (b) peroxidase is a "specific" secretory protein of the estrogen-sensitized uterine endometrium; (c) peroxidase synthesis is not a nonspecific response to steroid hormone action, since progesterone and testosterone do not induce its synthesis; (d) endogenous peroxidase is a possible diagnositc protein for the detection of estrogen-dependent growing tissues, including breast cancer; (e) movement of exogenous horseradish peroxidase from the interstitium to the uterine lumina is restricted by tight junctions located at the apices of epithelial cells. Estrogen and antagonists do not appear to influence the transepithelial movement of exogenous peroxidase into the lumen.

Estrogen and antagonist-induced changes in endometrial topography of immature and cycling rats.

The topographical changes of the luminal surface of the endometrium of immature and ovariectomized rats treated with estrogen, antagonists to estrogen, and progesterone. and during various stages of the estrous cycle and in pregnancy were examined by scanning electron microscopy. Massive increases in numbers and length of endometrial cell microvilli were observed at estrus, after injection of estradiol-17beta, diethylstilbestrol, estrogen plus progesterone. or the inhibitor C1628 to immature and ovariectomized rats. Withdrawal of the estrogen stimulus results in diminution of microvilli, producing a state identical to diestrus, during pregnancy, and after injection of progesterone, The estrogen antagonist appears to have both estrogenic and progestogenic properties, stimulating endometrial cell hypertrophy, secretion of protein, and production of numerous apical microvilli.

Parameter oscillation attenuation and mechanism exploration for continuous VHG ethanol fermentation.

A bioreactor system composed of a stirred tank and three tubular bioreactors in series was established, and continuous ethanol fermentation was carried out using a general Saccharomyces cerevisiae strain and a very high gravity medium containing 280 g L(-1) glucose, supplemented with 5 g L(-1) yeast extract and 3 g L(-1) peptone. Sustainable oscillations of glucose, ethanol, and biomass were observed when the tank was operated at the dilution rate of 0.027 h(-1), which significantly affected ethanol fermentation performance of the system. After the tubular bioreactors were packed with 1/2'' Intalox ceramic saddles, the oscillations were attenuated and quasi-steady states were achieved. Residence time distributions were studied for the packed bioreactors by the step input response technique using xylose as a tracer, which was added into the medium at a concentration of 20 g L(-1), indicating that the backmixing alleviation assumed for the packed tubular bioreactors could not be established, and its contribution to the oscillation attenuation could not be verified. Furthermore, the role of the packing's yeast cell immobilization in the oscillation attenuation was investigated by packing the tubular bioreactors with packings with significant difference in yeast cell immobilization effects, and the experimental results revealed that only the Intalox ceramic saddles and wood chips with moderate yeast cell immobilization effects could attenuate the oscillations, and correspondingly, improved the ethanol fermentation performance of the system, while the porous polyurethane particles with good yeast cell immobilization effect could not. And the viability analysis for the immobilized yeast cells illustrated that the extremely lower yeast cell viability within the tubular bioreactors packed with the porous polyurethane particles could be the reason for their inefficiency, while the yeast cells loosely immobilized onto the surfaces of the Intalox ceramic saddles and wood chips could be renewed during the fermentation, guaranteeing their viability and making them more efficient in attenuating the oscillations. The packing Raschig rings without yeast cell immobilization effect did not affect the oscillatory behavior of the tubular bioreactors, further supporting the role of the yeast cell immobilization in the oscillation attenuation.

Osmiophilic polymer generation: catalysis by transition metal compounds in ultrastructural cytochemistry.

A transition metal compound that is bound in tissues by any appropriate cytochemical reaction may catalyze the generation of an insoluble osmiophilic polymer from organic monomers such as 3,3'-diaminobenzidine. When the polymers are treated with osmium tetroxide, electron-opaque, insoluble osmium blacks (coordination polymers of osmium) are formed at the sites of the particular macromolecule or enzyme permitting its light, and electron, microscopic localization. This approach represents a distinct advantage over earlier cytochemical methods because the shorter incubation time needed here results in less artifactual deposition of metal ions, and less tendency to crystallize the reaction product. In addition, the shorter incubation times permit longer fixation of tissues and hence less artifact due to enzyme diffusion.

Heavy elements in surface materials: determination by alpha particle scattering.

The backscattering of alpha particles from a radioactive source can be used to determine the amounts of heavy elements such as lead in surface materials. A light, portable instrument has been constructed that can be used as a survey meter for painted surfaces. It has a sensitivity of 0.3 percent by weight in a measurement of a few minutes.

Ethanol fermentation technologies from sugar and starch feedstocks.

This article critically reviews some ethanol fermentation technologies from sugar and starch feedstocks, particularly those key aspects that have been neglected or misunderstood. Compared with Saccharomyces cerevisiae, the ethanol yield and productivity of Zymomonas mobilis are higher, because less biomass is produced and a higher metabolic rate of glucose is maintained through its special Entner-Doudoroff pathway. However, due to its specific substrate spectrum as well as the undesirability of its biomass to be used as animal feed, this species cannot readily replace S. cerevisiae in ethanol production. The steady state kinetic models developed for continuous ethanol fermentations show some discrepancies, making them unsuitable for predicting and optimizing the industrial processes. The dynamic behavior of the continuous ethanol fermentation under high gravity or very high gravity conditions has been neglected, which needs to be addressed in order to further increase the final ethanol concentration and save the energy consumption. Ethanol is a typical primary metabolite whose production is tightly coupled with the growth of yeast cells, indicating yeast must be produced as a co-product. Technically, the immobilization of yeast cells by supporting materials, particularly by gel entrapments, is not desirable for ethanol production, because not only is the growth of the yeast cells restrained, but also the slowly growing yeast cells are difficult to be removed from the systems. Moreover, the additional cost from the consumption of the supporting materials, the potential contamination of some supporting materials to the quality of the co-product animal feed, and the difficulty in the microbial contamination control all make the immobilized yeast cells economically unacceptable. In contrast, the self-immobilization of yeast cells through their flocculation can effectively overcome these drawbacks.

Photocatalytic reduction of nitrate in wastewater using ZnO nanopowder synthesized by solution combustion method.

ZnO nanopowder was synthesized by a unique method which is called solution combustion method (SCM). This nanopowder was used for a photocatalyst to decompose nitrate that is a toxic pollutant in wastewater. It has been known that TiO2, the most popular photocatalyst, does not decompose the nitrate. In this paper, however, the SCM ZnO nanopowder decomposed about 13% of nitrate. Furthermore, adding methanol as a hole scavenger, the decomposition rate was enhanced by about 5 times. On the other hand, it has been reported that the photocatalytic reduction reaction of nitrate produces ammonia as a final product. The present results, however, suggest that the final product is non-toxic nitrogen gas rather than the toxic ammonia. These results would be very valuable for drinking water purification.

Identification, subcellular localization, and estrogen regulation of peroxidase in 7,12-dimethylbenz(a)anthracene-induced rat mammary tumors.

An estrogen-induced, intensely staining peroxidase 3,3-diaminobenzidine-positive reaction product is found to be characteristic of hormone-dependent, 7,12-dimenthylbenz(a)anthracene-induced mammary tumors of the rat. This product is demonstrated in thick sections of such tumors from intact or estrogen-treated castrate rats but is not seen in tumors that are in regression due to castration or estrogen deprivation. It is, furthermore, absent from tumors whose growth is unaffected by castration. The subcellular localization of this enzyme activity is restricted mainly to the nuclear envelope and cisternae of the granular endoplasmic reticulum in addition to secretory granules. This provides the first evidence for a criterion that would allow differentiation of hormone-dependent and hormone-independent mammary cancer on histological sections and, as such, may have considerable potential as an aid in the classification of human breast cancer.

Studies on the antitesticular action of DL-6-(N-2-pipecolinomethyl)-5-hydroxy-indane (PMHI) in the rat.

Both prepubertal and adult rats were treated with a single oral dose of either 60 mg or 120 mg of dl-6-(N-pipecolinomethyl)-5-hydroxy indane maleate (PMHI) per kg of body weight. Their testicular weights were drastically reduced compared with those of the controls. A follow-up, beginning on the third day post-treatment and continuing for a period of 50 days, showed that the body weight growth of PMHI-treated rats was not retarded. The hormonal profile indicated that, except for FSH which showed a transitory elevation in PMHI-treated immature rats, the serum levels of LH, estrogen, and testosterone were indistinguishable from those of the controls. Testicular histology revealed that the spermatogenic process in PMHI-treated rats recovered at a dose-related rate. EM sections of testes of adult rats indicated that cytoplasmic vacuolation appeared in the Sertoli cells 5 h post-treatment. The consequent cascade of arrested spermiogenesis included abnormal acrosomal condensation of spermatids and sloughing of polynucleated spermatids. Some spermatocytes also seemed to be affected, but spermatogonia and Leydig cells remained intact. These results indicate the PMHI acts primarily on Sertoli cells and causes arrest in the spermiogenetic stage of the spermatids. At a higher and toxic dose of PMHI, however, the earlier germinal elements might also be affected, due to the extensive damage to the supporting Sertoli cells.

Mammalian endogenous peroxidases as cellular markers and as biosynthetic endpoints of hormone-mediated activity: viewpoint from cytochemistry.

Aldehyde-resistant, diaminobenzidine-stained endogenous peroxidases form ideal markers for the biochemical endpoints of hormone stimulation and differentiation of certain mammalian cells and tissues. The lactoperoxidase (LPO)-type of endogenous peroxidases are synthesized by the acinar cells of the salivary, Harderian, lacrimal and mammary glands and are present in their secretions. These LPO-type enzymes, that are inhibited by cyanide and aminotriazole, appear to operate extracellularly as bactericidal agents in milk and in other biological fluids. In the mammary gland, lactoperoxidase is a consistent marker enzyme for differentiated acinar cells engaged in lactogenesis. Myeloperoxidase (MPO)-type endogenous peroxidases are prominent markers for the GERL endomembrane system and differentiated lysosomes in certain cells of the reticuloendothelial system and phagocytes. MPO is prominent within eosinophils, peritoneal macrophages and in Kupffer cells. The MPO-type endogenous peroxidases function primarily within lysosomes as bactericidal agents. Thyroid peroxidase (TPO) is relegated to the cisternae of the granular endoplasmic reticulum and Golgi apparatus, to apical cytoplasmic vesicles and to the luminar cell membrane surface of acinar cells. The enzyme is probably activated at release and functions both in the organification reaction (T leads to To) and in the biosynthesis of thyroxine. Thyroid stimulating hormone (TSH) appears to play a key role in the regulation of TPO levels and activity in the thyroid gland. Certain tissues displaying growth-dependency on estrogen (i.e., uterus, cervix, vagina and the DMBA-induced rat mammary tumor) synthesize and secrete endogenous peroxidase into their lumina. These enzymes serve as important marker proteins of estrogen action, in that they occur distal to the binding of estrogen to its receptor protein. Estrogen antagonists, particularly CI-628 (Parke-Davis) and Nafoxidine (Upjohn) that appear to function through the estrogen receptor mechanism, also induce synthesis of the reproductive tract endogenous peroxidase but inhibit growth of these tissues. Progesterone antagonizes the synthesis of the reproductive tract peroxidases and inhibits growth of the tissues as well, in part, through the reduction of the cytosol estrogen receptor protein. Endogenous peroxidase activity appears to represent a reliable marker for rodent breast cancer tissues displaying dependency for estrogen and is of potential interest as a diagnostic marker protein in human breast cancer. Rat uterine peroxidase (UP) has been investigated by microelectrophoretic techniques. The molecular weight of UP has been determined in the range of 100,000 by using polyacrylamide gradient gels in the absence and presence of nonionic and anionic detergents. The isoelectric point of UP is located between pH 4.5 and 5.9. Employing the two-dimensional combination of isoelectric focusing and gel gradient electrophoresis, UP was separated into two subunits, one having a molecular weight of 70,000, the other less than 20,000.

Time course of mossy fiber degeneration following pontine ablation in the rat.

The normal ultrastructure of mossy fiber terminals within the ansiform lobule of the rat was investigated in parasagittal sections of the cerebellar cortex. Four types of mossy fiber varicosities (simple and complex dispersed, simple and complex clustered) were distinguished on the basis of the presence or absence of clublike excrescences and the concentration of synaptic vesicles within the cortical zone of the terminal. Removal of the pontocerebellar input to the contralateral cerebellar hemisphere was performed electrolytically. Such lesions, when placed in the basilar pontine gray using a dorsal approach, resulted in the degeneration of nearly the entire population of mossy fiber varicosities within the contralateral ansiform lobule. As early as 3 days following pontine ablation, degenerative changes were observed within the synaptic portions of the mossy fibers. Two distinct axonal reactions were apparent within this population of degenerating varicosities. A small population of mossy fiber varicosities (12%) of the simple clustered variety underwent a rapid course of electron-dense degeneration, which was complete by the fifth day. These mossy fiber varicosities were very susceptible to phagocytosis by reactive glial elements. The second group which consisted of simple (60%) and complex (26%) varieties of dispersed terminals, underwent a very slow course of electron-dense degeneration. Reactive glial cells were rarely found in association with this second group of degenerating varicosities. As a result, most glomeruli were found to contain debris from degenerating mossy fiber varicosities throughout the first 57 days following pontine lesions. The majority of the cerebellar glomeruli, however, were denervated by 80 days.