The effects of neem oil (Azadirachta indica A. JUSS) enriched with different concentrations of azadirachtin on the integument of semi‐engorged Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) females

Several studies searching for methods to control Rhipicephalus sanguineus s.l., (dog tick) infestations have been developed aiming to minimize the damages caused by these ectoparasites to the hosts and the environment, which is harmed by the indiscriminate use of toxic acaricide products. In this scenario, neem oil has been used as a natural alternative against ticks, once this chemical has repellent properties and interferes in the growth regulation of these ectoparasites, inhibiting ecdysis. The present study evaluated the effects of azadirachtin‐enriched neem oil on the integument of semi‐engorged R.sanguineus s.l., females through morphohistological techniques. The results showed the occurrence of significant morphological and histochemical alterations, mainly in the females exposed to higher concentrations, which demonstrates the dose‐dependent action of the chemical. A decrease in the cuticle thickness was observed, as well as a modification in the distribution of the epithelial cells, which displayed pyknotic and fragmented nuclei, and intensely vacuolated cytoplasm, indicating that these cells would be undergoing death processes. These morphological alterations observed in the integument of the females exposed to the azadirachtin‐enriched neem oil encourage the use of this chemical as a strategy to control these ectoparasites.     

Morphological alterations in the synganglion and integument of Rhipicephalus sanguineus ticks exposed to aqueous extracts of neem leaves (Azadirachta indica A. JUSS)

Currently, the necessity of controlling infestation by ticks, especially by Rhipicephalus sanguineus, has led researchers and public health managers around the world to search for new and more efficient control methods. This way, we can highlight neem (Azadirachta indica A. Juss) leaf, bark, and seed extracts, which have been very effective on tick control, and moreover causing less damage to the environment and to the host. This study showed the potential of neem as a control method for R. sanguineus through morphological and morphometric evaluation of the integument and synganglion of females, in semiengorged stage. To attain this, routine techniques of optical microscopy, scanning electron microscopy and morphometry of the cuticle and subcuticle of the integument were applied. Expressive morphological alterations were observed in both organs, presenting a dose‐dependent effect. Integument epithelial cells and nerve cells of the synganglion showed signs of cell vacuolation, dilated intercellular boundaries, and cellular disorganization, alterations not previously reported in studies with neem. In addition, variations in subcuticle thickness were also observed. In general, the effects of neem are multiple, and affect the morphology and physiology of target animals in various ways. The results presented in this work are the first evidence of its effects in the coating and nervous system of ticks, thus allowing an indication of neem aqueous extracts as a potential control method of the brown dog tick and opening new perspectives on acaricide use. Microsc. Res. Tech. 77:989–998, 2014. © 2014 Wiley Periodicals, Inc.     

Action of the insect growth regulator fluazuron,the active ingredient of the acaricide Acatak®, in Rhipicephalus sanguineus nymphs (Latreille, 1806) (Acari: Ixodidae)

The present study evaluated the efficacy of fluazuron (active ingredient of the acaricide Acatak^®) and its effects on Rhipicephalus sanguineus nymphs fed on rabbits exposed to different doses of this insect growth regulator. Three different doses of fluazuron (20 mg/kg, 40 mg/kg, and 80 mg/kg) were applied on the back of hosts (via “pour on”), while distilled water was applied to the Control group. On the first day of treatment with fluazuron (24 h), hosts were artificially infested with R. sanguineus nymphs. Once fully engorged, nymphs were removed and placed in identified Petri dishes in Biochemical Oxygen Demand (BOD) incubator for 7 days. After this period, engorged nymphs were processed for ultramorphological analysis. The results revealed alterations in the ultramorphology of many chitinous structures (smaller hypostome and chelicerae, less sclerotized scutum, fewer sensilla, fewer pores, absence of grooves, marginal and cervical strips and festoons in the body, even the anal plaque was damaged) that play essential roles for the survivor of ticks and that can compromise the total or partial development of nymphs and emergence of adults after periodic molting. Our findings confirm the efficacy of fluazuron, a more specific and less aggressive chemical to the environment and human health, and that does not induce resistance, in nymphs of the tick R. sanguineus in artificially infested rabbits treated with this arthropod growth regulator (AGR), indicating that it could be used in the control of this stage of the biological cycle of the tick R. sanguineus. Microsc. Res. Tech. 76:1177–1185, 2013. © 2013 Wiley Periodicals, Inc.     

Action of andiroba oil (Carapa guianensis) on Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi‐engorged females: Morphophysiological evaluation of reproductive system

Because of the increasing medical‐veterinary importance of ticks, the development of alternative control methods, less aggressive to the host and the environment has become the target of several researches. In this sense, the present study analyzed the action of different concentrations (5, 10, and 20%) of andiroba seed oil (Carapa guianensis) on the reproductive system of Rhipicephalus sanguineus females, through histochemical techniques and the quantification of the reproductive efficiency index. The results showed that andiroba oil is a potent natural agent, able to cause several changes in the oocytes of this species, impairing the reproductive success, once this natural product induces great physiological changes in the oocytes in all development stages, such as drastic reduction in proteins, polysaccharides, and lipids in these cells, and these components are essential for the viability of the embryo. In addition, it was observed that this product stimulate the oviposition, mainly at the concentration of 20%. This higher production of eggs represents a defense mechanism developed by the organism in order to ensure the reproductive success of the species, even in the presence of the toxic agent. However, the results obtained suggested that the laid eggs would not be viable, due to the great changes undergone by the oocytes. Thus, the present study showed that the use of this vegetal product would be an alternative way to control the ticks, bringing benefits similar to the ones obtained through the use of synthetic acaricides; however, with less damage to nontarget organisms and the environment as well. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

Morphological characterization of the nymphs Rhipicephalus sanguineus ticks (Latreille, 1806) (Acari: Ixodidae). Description of the testes, integument, Malpighian tubules, and midgut on the detachment day

This study presents the morpho-histological and histochemical characterization of the testes, integument, Malpighian tubules, and midgut of engorged Rhipicephalus sanguineus nymphs on the detachment day, showing the morphological and physiological characteristics to this phase in the life cycle of these individuals. The testis is constituted by germinative cells (only spermatogonia) with large, round-shaped and strongly stained nuclei which are organized into cysts by a thin layer of somatic cells. The integument consists of a cuticle subdivided into epicuticle (lipoprotein) and procuticle (glycoproteic), and a layer of epithelial cells which present glycolipoprotein elements. The procuticle presents two distinct regions: the exocuticle (next to the epicuticle) and the endocuticle (next to the epithelial layer). The Malpighian tubules present a simple epithelium with small flat and/or cubic cells, which form its wall and delimitates a lumen full of lipoprotein material. The midgut consists of an epithelial wall formed by two types of digestive cells, spent cells and empty digest cells, and by generative cells supported by a basal lamina and a thin layer of muscular tissue. This study described the main organs of engorged nymphs of R. sanguineus, to generate information that can help researchers to better understand the biology of these ectoparasites; which is fundamental for the development of compounds that are less aggressive to the environment. In addition, if the immature stages of the ticks are controlled, the number of adult ticks able to cause damages to the animals--and to the man as well--is also under control.     

Cytotoxic effects of extract of Acmella oleraceae (Jambú) in Rhipicephalus microplus females ticks

The present study analyzed the effects of different concentrations of the hexane extract of A. oleraceae (HEAO) (Jambú) on the germ cells of semi‐engorged Rhipicephalus microplus female ticks, through a morpho‐histological study, evaluating the effectiveness of the extract in the genesis of the individuals. To perform this analysis, 100 semi‐engorged females were divided into five groups with 20 individuals each: groups I and II, respectively constituted by distilled water control and 50% ethanol + 1% DMSO, and groups III, IV, and V constituted by treatment with HEAO in the concentrations of 12.5, 25.0 and 50.0 mg/mL, respectively. All the ticks were immersed in the different concentrations of the extract or in distilled water for 5 minutes, dried and conditioned in BOD incubator for 7 days. The individuals of the treatment groups revealed the action of this extract showing alterations in the germ cells of the females from the different groups when compared with those from the groups I and II (control groups). These alterations were mainly related to the size and shape of the oocytes; number of yolk granules; presence, number, size and location of vacuoles in the cytoplasm of all the germ cells; and the presence of nuclear alterations in these cells as well. Thus, it was demonstrated that the concentrations of HEAO affected the germ cells of R. microplus ticks. The effects of the extract are similar to those caused by renowned and efficient chemical products used to control these ticks. Microsc. Res. Tech. 79:744–753, 2016. © 2016 Wiley Periodicals, Inc.     

Microscopic comparative study of the exposure effects of testosterone cypionate and ethinylestradiol during prenatal life on the prostatic tissue of adult gerbils

There is an increasing variety of endocrine disrupting chemicals (EDCs) either with (anti)estrogenic or (anti)androgenic potential widely present in the environment. These xenosteroids may mimic endogenous steroid hormones disrupting the homeostasis of physiological pathways and leading to several disturbances, especially in tissues highly dependent on steroid hormones such as the prostate. Taking this into account, this comparative study aimed to verify the potential of ethinylestradiol (EE) and testosterone acting as ECDs on the prostate of both male and female adult gerbils exposed to these agents during the embryonic phase. Consequently, pregnant gerbils were treated either with 10 μg/kg/day of EE or with a single dose of 1 mg of testosterone cypionate. The pups that were born 6-8 days after testosterone exposure and the pups that were born after 3 days of EE exposure were allowed to grow but were sacrificed within 4 months. Serological, morphological, stereological, and immunohistochemical analyses were used. Overall, the results showed that both sexes exposed to testosterone and EE during gestation had a prostatic gland with an increased stromal and epithelial and a reduced luminal compartment. Moreover, we observed that glands affected with prostatic intraepithelial neoplasia showed intense stromal reshuffling. In conclusion, although these alterations were observed in both sexes, more relevant to this study was the differential responsiveness of males and females exposed to these different drugs. Whereas the EE affected males more, the testosterone was more harmful to the females.     

Sewage sludge toxicity in edaphic organism: analysis of midgut responses in the diplopod Rhinocricus padbergi

Diplopods are agile saprophagous invertebrates with the ability to colonize several soil layers. They feed on waste and decaying organic matter, making the soil in which they are present better aerated and mineral‐enriched. Because of their habits, diplopods have been suggested to be used in studies to analyze possible toxic effects of impacted soils in edafic fauna. In that respect, this study aimed at checking the feasibility of using morphological analysis from millipede's midgut as a biomarker for soils exposed to sewage sludge from sewage treatment plants. Histological and histochemical techniques, applied to the species Rhinocricus padbergi's midgut, were used. Two sewage sludge samples were used; they were collected in two small towns in the countryside of São Paulo State. Both cities are part of the Piracicaba–Capivari–Jundiaí basin (PCJ‐1 and PCJ‐3). For 7 and 15‐day periods, the animals were exposed to 10% concentrations and crude sludge for the PCJ‐1sample and to 1, 10, and 50% concentrations and sludge for the PCJ‐3 sample. The material exposed to the PCJ‐1 sample showed an increase in the presence of cytoplasmic granules in both concentrations in all periods and a slight increase in the rate of epithelial renewal. The material of PCJ‐3 sample presented the same alterations observed in PCJ‐1 sample in higher frequence and over that high quantity of neutral polysaccharides in the hepatic cells layer. We can conclude that with the increase of sewage sludge concentrations tested, there is an increase in morphophysiologicals alterations frequencies found in the diplopods' midgut. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

The occurrence of autophagic cell death in the tegument of rabbits pre‐infested with Rhipicephalus sanguineus and exposed to selamectin (active principle of acaricide pfizer revolution®)

Ticks of Rhipicephalus sanguineus species have great medical and veterinary importance for being a vector of various diseases. In an attempt to minimize their action on the host, people have resorted to chemical control by using various acaricides, such as selamectin. Although previous studies have demonstrated its toxic action in domestic animals, no studies focused on the detection of cell death when exposed to selamectin. For this reason, the technique for detecting autophagic cell death was used in order to demonstrate the responses of rabbits' skin tissues pre‐infested with R. sanguineus and exposed to different concentrations of selamectin. The obtained results when exposed to 100 and 80% concentrations of selamectin showed a strong mark of acid phosphatase on the cells of the connective tissue of the dermis and hair follicles, whereas the ones exposed to the 50% concentration had a weak mark on the cells of the connective tissue of the dermis and moderate staining in hair follicles. It became clear that, when used at high concentrations (100 and 80%), selamectin is capable to induce a large scale occurrence of the autophagic cell death process. On the other hand, the concentration of 50% causes minor morphophysiological changes in the skin of rabbit hosts when evaluated the cell death process. Therefore, the data confirms that selamectin is a powerful dose‐dependent toxic agent causes increased activity of the enzyme acid phosphatase. Microsc. Res. Tech. 76:1171–1176, 2013. © 2013 Wiley Periodicals, Inc.     

Morphological alterations of radicular dentine pretreated with different irrigating solutions and irradiated with 980-nm diode laser

Background: The topographical features of intraradicular dentine pretreated with sodium hypochlorite (NaOCl) or ethylenediamine tetraacetic acid (EDTA) followed by diode laser irradiation have not yet been determined. Purpose: To evaluate the alterations of dentine irradiated with 980‐nm diode laser at different parameters after the surface treatment with NaOCl and EDTA. Study design: Roots of 60 canines were biomechanically prepared and irrigated with NaOCl or EDTA. Groups were divided according to the laser parameters: 1.5 W/CW; 1.5 W/100 Hz; 3.0 W/CW; 3.0 W/100 Hz and no irradiation (control). The roots were splited longitudinally and analyzed by scanning electron microscopy (SEM) in a quali‐quatitative way. The scores were submitted to two‐way Kruskal–Wallis and Dunn's tests. Results: The statistical analysis demonstrated that the specimens treated only with NaOCl or EDTA (control groups) were statistically different (P < 0.05) from the laser‐irradiated specimens, regardless of the parameter setting. The specimens treated with NaOCl showed a laser‐modified surface with smear layer, fissures, and no visible tubules. Those treated with EDTA and irradiated by laser presented absence of smear layer, tubules partially exposed and melting areas. Conclusions: The tested parameters of 980‐nm diode laser promoted similar alterations on dentine morphology, dependent to the type of surface pretreatment. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.     

Toxicity of trifluralin herbicide in a representative of the edaphic fauna: Histopathology of the midgut of Rhinocricus padbergi (Diplopoda)

The aim of this study was to use the diplopod Rhinocricus padbergi as test organism to verify the possible toxic effects of the herbicide trifluralin in a representative of the edaphic fauna. Histological and histochemical techniques were applied in the midgut of the animals after exposure to different concentrations of trifluralin for 7 and 90 days (acute and subchronic exposure, respectively). The individuals from the control group did not present alterations to any of the exposures. In the acute exposure, the group exposed to a quarter and half of the field dose presented an accumulation of cytoplasmic granules in the hepatic cells; the field dose group presented an increase in the rate of epithelial renewal and the group exposed to double the field concentration presented an increase in the release of secretory vesicles. The subchronic response was obtained with the field dose and double field dose because the animals from a quarter and half groups did not survive until the end of the experiment. In both groups, the animals presented disruption of the epithelium and higher occurrence of hemocytes among the hepatic cells. The group exposed to double the field dose presented sites of cytoplasmic vacuolization, dilatation of the intercellular space, increase in the epithelial renewal, and release of secretory vesicles. The results alert for a careful use of this herbicide, since it promoted alterations in the midgut cells of the test organism used. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Inhibitory action of neem aqueous extract (Azadirachta indica A. Juss) on the vitellogenesis of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks

The present study revealed unheard of data about the action of aqueous extracts of neem leaves (Azadirachta indica) on the vitellogenesis of Rhipicephalus sanguineus ticks, proving that these extracts in 10 and 20% concentrations do not have the potential to kill the females; however, in lower concentrations (10%) provokes great morphological alterations in germinative cells such as the emergence of extended cytoplasmic vacuolization areas as well as the fragmentation of the germinal vesicle, even in those oocytes which were in initial stages of development (I-III), showing that neem is a potent agent which acts impeding one of the main metabolic stages of the ticks, i.e., the reproduction. In oocytes in final stages of development (IV-V) azadirachtin (neem's active principle) caused significant reduction in the size and quantity of proteic granules of the yolk and the inversion of their localization where the smaller granules before inside the cell (normal oocyte) were posteriorly observed in the periphery, and the bigger ones in the central region. Thus, the study showed that the alterations found both in the oocytes and in the pedicel cells indicated that azadirachtin acts on the process of tick's reproduction and signalizes that this plant can be used in the future to control ticks with the advantage of not being aggressive to nontarget organism or the environment. Furthermore, data here obtained showed that the most significant efficiency of the aqueous extract of neem is related to the concentration of 10%, proving that higher doses would not be so efficient.     

Histopathological lesions and toxicity in common carp (Cyprinus carpio L. 1758) induced by copper nanoparticles

Different types of metal oxide nanoparticles (NPs) have been suggested for various applications such as water treatment and construction of agricultural pesticides; however, there are concerns about the potential toxicity of these compounds for the nontarget organism especially aquatic organisms. The aims of this study were assessing toxicity and histopathological effects of copper oxide NPs (NPs‐CuO) on common carp (Cyprinus carpio) as a model organism. For this purpose 150 common carp with an average weight 7 ± 1 g were exposed to 0, 10, 20, 30, 40, 60, 80, 100, 150, and 200 mg/l of CuO‐NPs (10 treatment with three replicates) for 96 hrs. After 24, 48, 72, and 96 hrs exposures, mortality rates recorded and gill samples were collected. Statistical analysis showed significant differences in carp survival between control and treatment groups (p < 0.05); regression between fish mortality rate and NPs‐CuO concentration was also revealed (p < 0.01). The LC₅₀ 96h of NPs‐CuO for common carp was estimated as 124.9 mg/l in this study. Various tissue damages were observed in gill of treatments; such as, hypertrophy, hyperplasia, lamellar fusions, erythrocyte infiltration, epithelial lifting; also, there was significant correlation between intensity of tissue lesions and concentration of NPs‐CuO (p < 0.01). The findings of the present study demonstrate that sublethal concentration of NPs‐CuO can lead to serious tissue lesions. Whats more, concentrations above 30 ml/l of NPs‐CuO can lead to some clinical signs; such as skin darkening and death with open mouth as well as definite fish death.     

Effect of super short pulse Er:YAG laser on human dentin—Scanning electron microscopy analysis

This study evaluated the effect of different pulse widths in the morphological characteristics of human dentin irradiated with Er:YAG in cavity preparation protocols and dentin pretreatment. Dentin discs with 2 mm thickness were obtained from 18 human molars. The experimental groups were composed from two variables: (1) clinical protocol—cavity preparation (E = 200 mJ/20 Hz)—and pretreatment (E = 80 mJ/2 Hz); and (2) pulse duration—50, 300, and 600 μs. This formed six experimental groups (n = 3): G1 (E = 200 mJ/20 Hz/50 μs); G2 (E = 200 mJ/20 Hz/300 μs); G3 (E = 200 mJ/20 Hz/600 μs); G4 (E = 80 mJ/2 Hz/50 μs); G5 (E = 80 mJ/2 Hz/300 μs); G6 (E = 80 mJ/2 Hz/600 μs). The samples were irradiated with the Er:YAG laser by noncontact mode at a focal distance of 7 mm from the target point under continuous water spray (60% water and 40% air). After the irradiation, they were processed for scanning electron microscopy (SEM). Morphological analysis showed an irregular dentin surface, absence of smear layer with opening of the exposure of dentinal tubules and protruding peritubular dentin—without indications of changes for all protocols used. Regardless of the analyzed experimental group, the dentin surface showed a microretentive morphology characteristic of ablation. The G1 and G4 showed a rougher surface when compared to other groups. Finally, we concluded that the pulse width can influence the morphological characteristics of the irradiated dentin tested in different clinical indications. The larger surface irregularity caused by regulation with less pulse width (50 µs) seems more appropriate to get a microretentive pattern necessary for successful adhesives restoration procedures. Microsc. Res. Tech. 78:472–478, 2015. © 2015 Wiley Periodicals, Inc.     

Histometric and histomorphologic comparison of combustion and ambustion using in vivo reflectance‐confocal microscopy

Background: When combustion and ambustion induce a superficial injury, they are summarized as superficial burns, regardless of the underlying cause. Reflectance‐confocal microscopy (RCM) allows noninvasive imaging of the human skin on morphological features. We hypothesized that combustion and ambustion have different histomorphological effects on the human skin. Methods: Superficial burns caused by combustion (CO‐group, five females, three males; aged 26.8 ± 14.2 years) and caused by ambustion (AM‐group, four females, four males; aged 28.1 ± 13.8 years) were evaluated 24 h after injury. The following parameters were obtained using RCM on injured and noninjured (control) site: horny layer thickness, epidermal thickness, granular cell size, basal layer thickness. Results: Compared with the controls (12.8 ± 2.5 μm), horny layer thickness decreased significantly to 10.6 ± 2.1 μm in the CO‐group, whereas it increased significantly to 17.8 ± 2.8 μm in the AM‐group. The epidermal thickness did not differ significantly in CO‐group (47.9 ± 2.1 μm) and AM‐group (49.0 ± 3.1 μm), however, both increased significantly compared with the controls (42.7 ± 1.6 μm). The basal layer thickness increased more in AM‐group (17.0 ± 1.2 μm) compared to CO‐group (15.4 ± 1.1 μm). Both differed significantly compared with their controls (13.9 ± 0.9 μm). The granular cell size increased significantly in both groups ompared to the controls (721 ± 42 μm), however, a significantly higher increase was observed in CO‐group compared to AM‐group (871 ± 55 μm vs. 831 ± 51 μm). Conclusions: RCM evaluates significant histomorphological differences in superficial burns caused by combustion and ambustion. The term “superficial burn” should consider the underlying cause and thus supplemented by the term “combustion” or “ambustion.” Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Cytotoxic effects of thiamethoxam in the midgut and malpighian tubules of Africanized Apis mellifera (Hymenoptera: Apidae)

Due to its expansion, agriculture has become increasingly dependent on the use of pesticides. However, the indiscriminate use of insecticides has had additional effects on the environment. These products have a broad spectrum of action, and therefore the insecticide affects not only the pests but also non‐target insects such as bees, which are important pollinators of agricultural crops and natural environments. Among the most used pesticides, the neonicotinoids are particularly harmful. One of the neonicotinoids of specific concern is thiamethoxam, which is used on a wide variety of crops and is toxic to bees. Thus, this study aimed to analyze the effects of this insecticide in the midgut and Malpighian tubule cells of Africanized Apis mellifera. Newly emerged workers were exposed until 8 days to a diet containing a sublethal dose of thiamethoxam equal to 1/10 of LC₅₀ (0.0428 ng a.i./l L of diet). The bees were dissected and the organs were processed for transmission electron microscopy. The results showed that thiamethoxam is cytotoxic to midgut and Malpighian tubules. In the midgut, the damage was more evident in bees exposed to the insecticide on the first day. On the eighth day, the cells were ultrastructurally intact suggesting a recovery of this organ. The Malpighian tubules showed pronounced alterations on the eighth day of exposure of bees to the insecticide. This study demonstrates that the continuous exposure to a sublethal dose of thiamethoxam can impair organs that are used during the metabolism of the insecticide. Microsc. Res. Tech. 77:274–281, 2014. © 2014 Wiley Periodicals, Inc.     

Qualitative analysis of smear layer treated with different etidronate concentrations: A scanning electron microscopy study

The aim of this study was to examine the smear layer removal capacity of various etidronate treatments. Seventy‐eight roots were instrumented up to apical size X4 and randomly divided into five treatment groups and a positive control group (n = 13). Groups were as follows: irrigation with 5 ml of sterile saline (control) for 3 min, 5 ml of 17% ethylenediaminetetraacetic acid (EDTA) for 3 min, 5 ml of 9% A1‐hydroxyethylidene‐1,1‐bisphosphonate (HEBP) for 3 min, 5 ml of 18% HEBP for 3 min, irrigation with 5 ml of 1% sodium hypochlorite (NaOCl) + 9% HEBP for 3 min, and 5 ml of 2% NaOCl + 18% HEBP for 3 min. The roots were examined using a scanning electron microscope. The smear layer scores in the 2% NaOCl + 18% HEBP group were lower than those in the other treatment groups and the control group (p < .05). There were no significant differences among the treatment groups in the smear layer scores of the coronal and middle thirds. However, the smear layer scores in the 2% NaOCl + 18% HEBP treatment group were lower those in the 9% HEBP and 18% HEBP treatment groups in the apical third (p < .05). In the clinical setting, the recommended concentration of HEBP is 18%, and it should be used with an oxidizing agent to ensure optimum smear layer removal.     

Observations of human corneal epithelium by tandem scanning confocal microscope

We applied the tandem scanning confocal microscope (TSCM) to 30 healthy human corneas of 3 normal volunteers and 27 patients with cataract and retinal detachment to observe normal corneal epithelial cells in vivo. All corneas were normal under slit lamp microscopic examination. The superficial and basal epithelial cells close to the basal lamina in the central cornea were recorded on videotape and analyzed by a computer-assisted digitizer. The mean cell areas of superficial cells exposed at the surface and basal cells at the horizontal section were 624 ± 109 μm² and 66 ± 5 μm², respectively. The ratio of superficial to basal mean cell area was 11.0 ± 4.5. TSCM was thus useful in evaluating the relationship between superficial and basal cells in human corneal epithelium in vivo.     

Observations of human corneal epithelium by tandem scanning confocal microscope

We applied the tandem scanning confocal microscope (TSCM) to 30 healthy human corneas of 3 normal volunteers and 27 patients with cataract and retinal detachment to observe normal corneal epithelial cells in vivo. All corneas were normal under slit lamp microscopic examination. The superficial and basal epithelial cells close to the basal lamina in the central cornea were recorded on videotape and analyzed by a computer-assisted digitizer. The mean cell areas of superficial cells exposed at the surface and basal cells at the horizontal section were 624 ± 109 μm² and 66 ± 5 μm², respectively. The ratio of superficial to basal mean cell area was 11.0 ± 4.5. TSCM was thus useful in evaluating the relationship between superficial and basal cells in human corneal epithelium in vivo.     

Do universal adhesives provide the benefits from double-application or an extra bonding layer in composite repair?

This study aimed to determine whether the application of extra hydrophobic resin (EHR) or double layer (DL) improves microtensile bond strength (μTBS) of two universal adhesives to composite resin. Composite blocks were fabricated and exposed to thermal cycles. The specimens were horizontally sectioned into two halves. Scotch Bond Universal (SBU) and Clearfil S3 Bond Universal (CSBU) were applied to the ground composite surface according to the manufacturers' instructions, or with DL application or EHR application. The repair composite was incrementally placed to bonded planes. Composite sticks were achieved and tensed with a universal testing machine, followed by examining the fracture surfaces by a scanning electron microscope. Data were evaluated by Weibull analysis (shape and scale [σ_θ and σ_0.10] parameters) using the maximum likelihood method. The σ_θ and σ_0.10, respectively, estimate strength at 63.2 and 10% probability of failure. Shape parameter values showed significant differences among treatments. DL application of CSBU did not affect μTBS values at σ_θ of failure but caused to decrease μTBS values at σ_0.10 of failure. DL application of SBU reduced μTBS values at σ_θ of failure. DL or EHR coating did not improve μTBS of SBU. EHR application increased μTBS of CSBU, whereas DL application did not benefit.