Effect of streptozotocin on the ultrastructure of rat pancreatic islets

Our objective was to study the effects of three (30, 40, and 50 mg/kg) doses of Streptozotocin (STZ) on fasting plasma glucose level (FPG) and observe its effects at the cellular level in rat pancreas by electron microscopy. FPG was measured in rats before induction of diabetes and then on 3, 7, and 14 days after induction of diabetes with STZ. Keto diastix urine strips were used to check urine glucose and ketone bodies. Two weeks after the induction of diabetes, the rat pancreas was removed and fixed for light and electron microscopic studies. Three days after induction, the mean FPG level was 112 mg/dl in Group I (30 mg/kg STZ), 217 mg/dl in Group II (40 mg/kg STZ), and 376 mg/dl in Group III (50 mg/kg STZ). Histology was normal in Group I but revealed altered islet structure in Groups II and III. Ultrastructure revealed intact D cells in all three groups. The focal mitochondria and Golgi complex swelling found in A and B cells was occasional in Group I and frequent in Groups II and III. Swelling of other organelles and reduction in the size and number of granules was further observed in Group III. It is our conclusion that the 30-mg/kg body weight STZ produces mild changes while 50 mg/kg proves to be fatal. STZ at 40 mg/kg has a moderate effect on plasma glucose as well as on the islets of Langerhans at a cellular level.     

Genotoxic and mutagenic effects of permethrin in mice: Micronuclei analysis in peripheral blood erythrocytes

Pyrethroids such as permethrin are synthetic compounds widely used in the agriculture of many countries to combat plagues and in domestic products, such as acaricides. Not so long ago these chemicals were characterized as non‐toxic for non‐target organisms; however, recent studies have showed that these compounds could present toxic potential for many organisms. In this sense, this study presents genotoxic and mutagenic potential of permethrin administered intraperitoneally in mice under artificial conditions by the use of micronucleus assay in the peripheral blood of these animals. The mice were divided into five groups: group I = negative control (distilled water), group II = positive control (cyclophosphamide), group III = 30% of permethrin LD₅₀ (96 mg/kg), group IV = 50% of permethrin LD₅₀ (160 mg/kg), and group V = 80% of permethrin LD₅₀ (256 mg/kg). The peripheral blood was collected 24, 48, and 72 h after treatment. Results showed that all the tested permethrin dosages presented genotoxic and mutagenic effects 24 h after treatment, which would contradict the classification of this chemical product as moderately toxic, i.e., unable to cause damages to the cell DNA. Microsc. Res. Tech. © 2012 Wiley Periodicals, Inc.     

Ketamine effects on the urogenital system--changes in the urinary bladder and sperm motility

Different doses of ketamine (10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, and 60 mg/kg) were injected i.p. (I.P.), respectively, to male ICR mice to determine the optimal dosage for chronic administration. At and above 40 mg/kg I.P. injection, mice had almost no hindlimb movement during swimming test. Subsequently, 30 mg/kg was used as the dose for the study in the toxicity of long-term ketamine administration on urinary bladder and sperm motility. The treatment group were subdivided into two (n = 10 each group); one received daily ketamine treatment i.p. for 3 months and another group for 6 months. Corresponding number of mice in control groups (n = 5 each group) received saline injection instead of ketamine. Terminal dUTP nick and labeling (TUNEL) study and Sirius red staining were carried out on the sectioned slides of the urinary bladders to study the degree of apoptosis in both epithelium and muscular layers of the urinary bladder and the relative thickness of the muscular layers in this organ was also computed. Apoptosis in the bladder epithelium was observed initially in the 3-month ketamine treated mice and the number of apoptotic cells was significantly different (P < 0.05) between the 3-month and 6-month ketamine treated mice and the control. The relative thickness of muscular layers in the bladder wall also decreased significantly (P < 0.05) when the 6-month treated mice and the control were compared. Sirius red staining revealed increase of collagen in the urinary bladder of the treated mice, most evidently 6 months after ketamine treatment. In addition, the sperm motility was studied and there was a statistically significant difference between the control and ketamine treated groups in the percentages of sperms which were motile (P < 0.05). This suggested that the chronic administration of ketamine affected the genital system as well.     

Ultrastructural changes in the cemento‐enamel junction caused by acidic beverages: An in vitro study

The present in vitro study was aimed at evaluating the morphological changes in the cemento‐enamel junction (CEJ) after exposure to acidic beverages using the scanning electron microscopy (SEM). The initial pH and titratable acidity (TA) was analyzed from follow groups: (I) Coca cola, (II) orange juice, (III) Cedevita, (IV) Red Bull, (V) Somersby cider, and (VI) white wine. The CEJ samples (n = 64), obtained from unerupted third molars, were allocated to one control (artificial saliva, n = 16) and six experimental groups (n = 8). The experimental samples were immersed in beverages (50 ml) for 15 min, three times daily, 10 days, and in artificial saliva between immersions. SEM analysis was performed in a blind manner, according to scoring scale. One‐way ANOVA and Tukey's post hoc tests, as well as Kruskal–Wallis and Mann–Whitney U test used for statistical analysis. The pH values of the acidic beverages ranged from 2.65 (Coca cola) to 3.73 (orange juice), and TA ranged from 1.90 ml (Coca cola) to 5.70 ml (orange juice) of NaOH to reach pH 7.0. The SEM analysis indicated statistically significant differences between the control samples and those immersed in acidic beverages. The Groups IV, I, and II, showed the highest CEJ damage grade while those of the Group VI were the lowest. All the tested acidic beverages caused morphological changes in the CEJ with a smaller or larger exposure of dentine surface, and were not always related to the pH or TA of acidic beverages.     

Fipronil (active ingredient of acaricide Frontline®) acting on the mice thyroid

Fipronil, active ingredient of Frontline® acaricide, is a phenyl‐pyrazolic derivative, and its efficacy in the elimination of several plagues, even in lower concentrations, has already been demonstrated; however, its effect on nontarget organisms has not been thoroughly explained. In this sense, the present study had the objective to evaluate the effects of different doses of fipronil in mice and determine its cytotoxic potential and possible compromises on nontarget organisms in artificial conditions. The results showed that animals exposed to fipronil presented significant alterations in the thyroidean tissue with evident follicular disorganization, decreased in the size of most follicles culminating with the rupture of smaller follicles and still with posterior fusion in a big and irregular “follicle” filled with colloid. The present study also revealed that the action of fipronil not only causes disorganization in the thyroidean tissue but alsoalters the chemical composition of the colloid itself (prehormone), modifying its proteic basis, which was confirmed by histochemical test for protein detection, making it clear that, in thyroids exposed to fipronil, the colloid was weakly positive or even negative for the presence of proteins in its constitution. Microsc. Res. Tech. 2011. © 2011 Wiley Periodicals, Inc.     

固相萃取和气相色谱法测定大米中氟虫腈(锐劲特)残留

建立运用固相萃取技术检测大米中的氟虫腈残留的气相色谱分析方法。样品经过乙腈/丙酮(70/30)抽提,通过硅酸镁(Florisil)固相萃取(SPE)小柱净化,旋转蒸干后用乙腈定容,应用微池电子捕获检测器(μ ECD)检测。样品在10 μ g/kg、15 μ g/kg和30 μ g/kg三个浓度水平添加,回收率为83.18%、 84.49%和88.85%,方法变异系数为1.60%、 1.89%和1.54%。本方法在大米样品中的最低检测定量限为10 μ g/kg     

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.     

Morphological and histochemical study of the masseter muscle after occlusal alteration

The aim of this study was to evaluate alterations in the masseter muscle of 30 male guinea-pigs submitted to occlusal alteration. The animals were divided into 2 equal groups, the control group (C) only submitted to surgical stress, and the occlusal altered group (T) submitted to teeth extraction. Each group was subdivided into 3 groups, with 5 animals, for the following studies: macroscopy and vessels distribution, light microscopy and histochemical analysis, with animals’ perfusion 2 months after surgery. Data were submitted to statistical analysis. Macroscopically, an anteroposterior orientation of the muscular fibers was found and statistical difference between C and T groups in width (p<0.05). Microscopically, C and T groups showed polygonal muscular fibers with variable diameters, and on the left side of the T group these differences were more pronounced. Histochemically, in both groups, the prevalence of intermediate reactivity fibers and several high reactivity fibers spread out in the deeper area was observed, with no significant differences between superficial fibers on both sides of C and T groups. It was possible to conclude that the masseter muscle in guinea-pigs was sensible to functional chewing alteration.     

Effects of anti-CD3 monoclonal antibody in salivary glands of spontaneously diabetic mice

Background: Diabetes mellitus results in many complications, also compromising the salivary glands. The current treatment for this condition should be a substituting method to exogenous insulin. In this aspect, the immunotherapy has been tested, but, it can be inefficient as an agent for the control of damage caused by diabetes. Thus, the aim of this study was to evaluate the anti‐CD3 monoclonal antibody as alternative immunotherapy in the recovery of salivary glands of spontaneously diabetic NOD (nonobese diabetic) mice. Methods: NOD mice were divided into two groups of 10 animals: group I (untreated diabetic mice) and group II (anti‐CD3‐treated diabetic mice). After treatment, the samples of salivary glands were collected for histological examination under both transmitted and polarized light microscopy. Results: Alterations in tissue architecture; increase in extracellular matrix and presence of inflammatory process were observed in untreated animals. Recovery of the salivary acinar cells occurred in treated animals. The parotid glands demonstrated a smaller amount of collagen fibers and were not observed severe inflammatory processes. Conclusion: These results indicate that immunotherapy contributed to reestablishment of tissue damaged by the hyperglycemic condition, demonstrating that the immunomodulation plays an important role in the recovery of salivary glands. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

高效液相色谱-串联质谱法测定稻渔土壤中氟虫腈及其代谢物残留

建立了高效液相色谱-串联质谱（HPLC-MS/MS）法测定稻渔土壤中氟虫腈及其代谢物残留。样品经乙腈恒温振荡提取,ENVI-18固相萃取柱和10 mg GCB净化,ZORBAX Extend-C18色谱柱（2.1 mm×100 mm×1.8 μm）分离,电喷雾负离子模式检测,基质匹配标准曲线外标法定量。结果表明,稻渔土壤中氟虫腈、氟甲腈、氟虫腈砜和氟虫腈亚砜的平均回收率在83.5%~110.3%之间,相对标准偏差在1.9%~11.5%之间,氟虫腈及其代谢物在0.5~100 μg/L浓度范围内的线性关系良好,相关系数均大于0.998,检出限和定量限分别为1.0、3.0 μg/kg。该方法操作简单、结果准确、可靠,可实现样品的批量化处理,能够满足稻田水产品中氟虫腈及其代谢物检测的需求。     

Structure and distribution of sensilla on maxillary palps and labella of Lutzomyia longipalpis (Diptera: Psychodidae) sand flies

The typology, number, size, and distribution pattern of labellar and palpal sensilla of male and female Lutzomyia longipalpis (Lutz and Neiva, 1912) sand flies were analyzed by scanning electron and light microscopy, contributing to the construction of a gustatory and olfactory map in this important vector species. About 35 trichoid sensilla, sorted into three different classes according to their lengths, are present on the two labellar lobes. Five distinct structures are observed in the maxillary palps, which could be classified as noninnervated (microtrichia and scales) or the following innervated structures: 1) campaniform sensilla, found near the distal end of palpal segments II and III; 2) multiporous capitate peg sensilla, found in reduced number (1-6) on segment II, but located in a group of 15-25 on the distal third part of segment III; and 3) trichoid sensilla, distributed linearly on segment IV and spirally on segment V. Light microscopy analysis of silver-stained specimens indicated the presence of pores in the trichoid sensilla and their chemosensory function is discussed, as well as the possible function of the other sensilla.     

Evaluation of the hepatoprotective and antioxidant effects of <i>Tegillarca granosa</i> flesh body extract against potassium bromide toxicity via targeting the histomorphometry,chromosomal and expressions of TGF-β1, VEGF and COX-2 genes in rats

The hepatotoxic effect of potassium bromide (KBr) on rat liver tissues were determined, as well as the potential protective effect of Tegillaraca granosa (T. granosa) flesh body extract. Twenty adult male albino rats were equally distributed into four groups; Group (I) treated with physiological saline (control group), Group (II) was orally gavaged by 200 mg/kg of T. granosa body extract day after day, Group (III) was intoxicated by KBr (150 mg/kg bwt day after day orally) and finally, Group (IV) was given a combination of T. granosa flesh body extract plus KBr with similar doses in the second and third groups. At the end of one month, blood, liver tissue and bone marrow samples were collected to be used for the required laboratory examinations. In response to KBr toxicity, there was a significant increase in serum antioxidant biomarkers, which was accompanied by a significant change in hepatocyte ultrastructure and a significant change in carbohydrate and protein levels within the liver organ. In addition, KBr intoxication resulted in a substantial increase in the incidence of chromosomal aberrations such as holes, splits, deletions, fragments, ploidy, and ring chromosomes, as well as significant upregulation of TGF-1, VEGF, and COX-2 gene expression. The hepatotoxic effect of KBr was counteracted by treatment with T. granosa flesh body extract. T. granosa flesh body extract has a curative antioxidant and numerous protective effects against KBr hepatotoxicity.     

Ameliorative effects of melatonin and zinc oxide nanoparticles treatment against adverse effects of busulfan induced infertility in male albino mice

Testicular damage is one of the most hazardous effects as it’s associated with azoospermia. Busulfan (Bu) is a highly toxic chemotherapeutic drug that affects testis. Thirty male Swiss albino mice divided into six groups of 5 animals each. Control (oral 0.9% saline daily for 75 days); Mel (20 mg/kg/day orally for 30 days); ZnO NPs (5 mg/kg/day i.p. for 30 days); BU (single i.p. injection of 40 mg/kg and then left for 45 days); BU + Mel (single 40 mg/kg dose of BU and left for 45 days followed by 20 mg/kg/day Mel for 30 days); BU + ZnO NPs (single dose of 40 mg/kg of BU and left for 45 days, then 5 mg/kg/day ZnO NPs for 30 days). Preparation and Characterization of ZnO NPs. Specimens from testis prepared for ultrastructural investigations using TEM after Masson’s trichrome and toluidine blue staining. BU induced histological and ultrastructural damage of the testis. Moreover, the present results could be concluded that Mel or ZnO NPs can protect the testicular tissue against ultrastructural alterations induced by BU by its antioxidant and anti-apoptotic effects.     

Role of resistance physical exercise in preventing testicular damage caused by chronic ethanol consumption in UChB rats

Ethanol consumption is associated with spermatogenesis damage and testosterone level alterations. Alcohol remains the most commonly used substance among athletes and sports enthusiasts. This study evaluated whether resistance physical exercise can reduce the testicular damage caused by ethanol exposure. A total of 36 ethanol drinking (UChB) rats were divided into four groups: C (control rats), ETOH (ethanol consumption), ETOH + T (ethanol consumption + physical training), and T (group physical training). The physical training component of the T and ETOH + T groups was based on a resistance training model consisting of four sets of 10 jumps, with an increasing overload of 50–70% of the body weight attached to the chest three times per week. Rats in the ETOH and ETOH +T groups received 10% ethanol. At postnatal day 90, the rats were sacrificed. Blood sample was collected for hormonal analysis, and the testicles were weighed and processed for histopathological, morphometric, and immunohistochemical analyses. The ETOH group showed an increase in testosterone levels. The immunohistochemical of androgen receptor and the absolute weight of the testes were higher in the ETOH and ETOH + T groups, while the ETOH animals showed a decreased weight gain index. The number of abnormal seminiferous tubules increased in the ETOH and T groups compared to those in the control group (C); however, the association with treatment (ETOH + T group) prevented this effect and decreased caspase‐3 production. In conclusion, these findings show that the combination of ethanol consumption and resistance physical exercise can prevent testicular damage in adult UChB rats.     

On Material Removal Regimes for the Shaping of Glass Edges: Force Analysis,Surface Topography and Damage Mechanisms

Glass shaping, which corresponds to the removal of the edges of a specimen, is the last finishing operation in glass manufacturing. This process has several functions on the final shaped glass including removing sharp edges, improving mechanical resistance, decreasing surface damage and giving it an aesthetical aspect. This article addresses the effects of working parameters, including grinding forces and consumed power, on surface edge finishing and damage mechanism induced during glass grinding. Microscopic observations and multi-scale analysis have also been conducted to investigate the surface edge characteristics. Experimental results show three damage regimes. The first (regime I) is a partial ductile regime with cutting action accompanied by chip formation. The second (regime II) is a crushing (or fragmentation) regime. The last (regime III) is also a partial ductile regime but by ploughing action with displaced material. The shaped surface obtained in the regime II has a better roughness than that obtained in regime I and III. However, regimes I and III include streaks and form defects which are not present in regime II. Similar to metallic materials, the evolution of force components show a linear relationship between normal and tangential forces. This implicates a constant average contact pressure and friction coefficient (μ) between the flat grains and the workpiece.     

Creep mechanisms of a new Ni-Co-base disc superalloy at an intermediate temperature

The microstructures of a new Ni-Co-base disc superalloy, TMW-4M3, before and after the creep test at 725 °C/630 MPa have been systematically investigated by transmission electron microscopy (TEM). The crept microstructures were marked as three different deformation stages (I, II and III) corresponding to the gradually increased strain. At stage I, stacking fault (SF) shearing was the main deformation mechanism. The SF was extrinsic and lay on {111} plane. However, deformation microtwinning became the dominant mode at stage II and III. The average spacing of deformation twins decreased from 109 ± 15 nm at stage II to 76 ± 12 nm at stage III, whereas the twin thickness did not change significantly. The influence of stacking fault energy (SFE) of γ matrix on the deformation mechanism is discussed. It is suggested that lower SFE in TMW-4M3 is partly responsible for the enhanced creep resistance.     

Cyclosporine improves remyelination in diabetic rats submitted to a gliotoxic demyelinating model in the brainstem

The use of cyclosporine (CsA) has shown to induce an increase in density of oligodendrocytes near remyelinating areas following the injection of ethidium bromide (EB), a demyelinating agent, in the rat brainstem. It is also known that diabetes mellitus was capable of delaying remyelination by both oligodendrocytes and Schwann cells in this gliotoxic model. This study was designed to assess whether CsA had the capacity to improve remyelination in streptozotocin‐induced (50 mg/kg, intraperitoneal route) diabetic rats. Diabetic Wistar rats were divided in different groups receiving 10 microlitres of 0.1% EB or 0.9% saline solution into the cisterna pontis and were treated or not with CsA. During 7 days and, thereafter, three times a week, 10 mg/kg/day of CsA were given by intraperitoneal route. The rats were euthanized from 7 to 31 days after EB or saline injection and brainstem sections were collected and processed for light and transmission electron microscopy studies. Results from different groups were compared by using a semi‐quantitative method developed for documenting the extent and nature of remyelination in semithin sections following gliotoxic lesions. Results showed that CsA administration to diabetic rats after EB injection stimulate both oligodendroglial and Schwann cell remyelination (mean remyelination scores of 3.15 ± 0.5 for oligodendrocytes and 1.36 ± 0.58 for Schwann cells) compared to untreated animals (2.52 ± 0.71 for oligodendrocytes and 0.73 ± 0.47 for Schwann cells, respectively). CsA given to diabetic rats was capable of reversing some of the deleterial effects of diabetes on remyelination. Microsc. Res. Tech. 76:714–722, 2013. © 2013 Wiley Periodicals, Inc.     

Evaluation of testicular tissue of adult rats treated with cisplatin incorporated into the liposome

Cisplatin (CPL) is one of the most widely used and effective chemotherapeutic agents for the treatment of several human malignancies. However, it causes serious side effects, especially on reproduction. In order to reduce the undesirable effects caused by many drugs, liposomes have been used as a good system for drug delivery. The aim of this study was to investigate, for the first time, the effects of CPL incorporated into the dipalmitoyl phosphatidylcholine liposome (DPPC) on the testicular tissue of adult Wistar rats. The animals (n = 20) were distributed into four experimental groups: (a) control (distillated water); (b) liposome (DPPC, 1 mL), (c) cisplatin incorporated into liposome (CPL/DPPC), and (d) CPL (8 mg/kg body weight). The animals received a single intraperitoneal injection and were killed 10 days after each treatment for histopathological analysis of testes. The results showed that the testicular histomorphometric parameters in rats of DPPC and CPL/DPPC groups were similar to those of the control group. Meanwhile, rats of the CPL‐treated group showed a variety of morphological alterations, including atrophy of seminiferous tubules and presence of multinucleated cells in the germinal epithelium. The incorporation of CPL into the liposome had no influence on the testicular weight or any other stereological parameters, but it was beneficial in maintaining the body weight of the animals. In conclusion, the liposome suppressed the cytotoxic effects caused by cisplatin in the testes of rats, suggesting a possible use in chemotherapy against cancer to reduce the side effects seen on reproduction. Microsc. Res. Tech. 78:323–329, 2015. © 2015 Wiley Periodicals, Inc.     

Testing of total hip replacements: endurance tests and stress measurements. Part 1: Endurance tests

This is a paper in two parts. Part 1 gives details of fatigue tests carried out on Howse II hip prostheses. Three types of tests were carried out in accordance with three different draft test standards. Standard (I) was BSI DD91: 1984 in which the load range is 0.3-4.1 kN and the antero-posterior offset angle is 15 degrees. Standard (II) was BSI DD91: 1986 where the load range is 0.3-2.3 kN and the offset angle is 9 degrees. Standard (III) was ISO/DP 7206/3 in which the load range is 0.3-4.1 kN and the offset angle is 0 degrees. Four specimens were tested according to each standard. It was found that all the specimens tested according to standard (I) failed by fatigue after less than 6 x 10(5) cycles. All the specimens tested according to standards (II) and (III) withstood 5 x 10(6) cycles without failure. It is clear that the offset angle has an important influence on the fatigue life of the prosthesis. It is concluded that tests according to standard (I) are too severe, while tests according to standards (II) and (III) are either adequate or lax. This led to Part 2 of the investigation where the stresses in the prostheses were measured and calculated.     

Influence of anti-inflammatory administration in collagen maturation process during orthodontic tooth movement

Bone formation is essential to orthodontic tooth movement and bone is formed by collagen. To analyze the collagen maturation process on bone matrix neoformed under nonsteroidal and steroidal treatment during orthodontic tooth movement by polarized microscopy, male Wistar rats (n = 90) were randomly divided into three groups (n = 30): C (control), NSAID (potassium diclofenac) and SAID (disodic phosphate dexamethasone). The animals of the C group received 0.9% saline solution; NSAID group received 5 mg/kg potassium diclofenac (CATAFLAM®); and SAID group received 2 mg/kg phosphate dissodic dexamethasone (DEXANIL®). Animals were sacrificed 3, 7 or 14 days after the placement of orthodontic appliances and the upper first molars were processed histologically and stained with picrosirius. Bone formation was evaluated under polarized light microscopy and 4.5 Image Pro‐Plus® software calculated the percentage of immature/mature collagen present in the groups. On the third days after force application, SAID and NSAID groups showed greater proportion of immature collagen than C group. On the seventh and fourteenth days, there was a lower proportion of mature collagen only in the SAID group (P < 0.001). These data demonstrate that dexamethasone delays the collagen maturation process in established bone matrix. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.