Alterations of C‐MYC,NKX3.1, and E‐cadherin expression in canine prostate carcinogenesis

The dog (canis lupus familiaris) is the only other species besides humans that develop spontaneous prostatic carcinomas (PCa) at a high frequency. The canine model is primarily utilized for the study of the PCa molecular mechanisms and provides a natural animal model for the study of potential therapies. In humans, the PCa frequently exhibits mutations in the C‐MYC and a reduced expression of the E‐cadherin and NKX3.1 proteins. This study's objective was to evaluate the NKX3.1, C‐MYC, and E‐cadherin expression in the canine normal prostate, benign prostatic hyperplasia (BPH), proliferative inflammatory atrophy (PIA) and PCa and to verify differences in expression and subcellular localization of these proteins in the prostatic carcinogenesis. A tissue microarray (TMA) slide was constructed, and immunohistochemistry with antibodies raised against C‐MYC, NKX3.1, E‐cadherin and p63 was performed using the peroxidase and DAB methods. The C‐MYC protein expression was elevated in the cytoplasm and nuclei of the canine PCa and PIA compared with the normal prostate (P = 0.004. The NKX3.1 protein expression was reduced in 94.75% of the PCa and 100% of the PIA compared with the normal prostate (P = 0.0022). In fact, the expression of E‐cadherin trended towards a decrease in carcinomas when compared to normal prostate and PIA. By immunohistochemistry, more p63‐positive basal cells were observed in the PCa and PIA when compared with the normal prostate (P = 0.0002). This study has demonstrated that the carcinogenesis of canine prostatic tissue may be related to basal cell proliferation, the gain of C‐MYC function and the loss of NKX3.1 protein expression. Microsc. Res. Tech. 76:1250–1256, 2013. © 2013 Wiley Periodicals, Inc.     

Oxidative effects of glyphosate on the lipophobic intracellular environment in the microalgae <i>Chlorella vulgaris</i>

The studied hypothesis is that the herbicide glyphosate (GLY) can affect the oxidative balance in the hydrophobic intracellular medium in non-target Chlorella vulgaris cells. Analytical GLY and RoundUp (RUP) supplementation, affected the growth profile. A significant 42% decrease in the cellular biomass in stationary (St) phase was observed in cultures supplemented with either 5 µM of GLY or RUP, as compared to control cultures. The treatment with 0.3 µM of GLY generated non-significant effects on the oxidation rate of 2’, 7’ dichlorofluorescein diacetate (DCFH-DA), neither in exponential (Exp) nor in St phase of development, as compared to control cultures. However, the treatment with either 5 µM GLY or 0.3 and 5 µM RUP lead to a significant decrease in the DCFH-DA oxidation rate, as compared to control cultures. The lipid radical (LR^●) generation rate, detected by Paramagnetic Resonance Spectroscopy (EPR), was significantly increased in the presence of RUP, in Lag and Exp phase of growth. The non-enzymatic antioxidants, α-Tocopherol (α-T) and β-Carotene (β-C), are aimed to protect membranes against the damage produced by the radical reactions. The content of β-C was not significantly affected, as compared to control cultures, by any of the treatments, in both growth phases of cellular development. The content of α-T was significantly decreased by the supplementation with either 0.3 or 5 µM of RUP or 5 µM GLY. The LR^●/α-T ratio, used as indicator of the oxidative balance in the hydrophobic cellular media, was significantly different between samples obtained from control and RUP-exposed microalgae in both, Exp and St phase of development, with either 0.3 or 5 μM RUP. The data presented here showed evidence that suggested that oxidative balance in the hydrophobic environment was affected by either GLY or RUP.

     

Synergistic combination of colistin with imipenem,amikacine or ciprofloxacin against <i>Acinetobacter baumannii</i> and <i>Pseudomonas aeruginosa</i> carbapenem-resistant isolated in Annaba hospital Algeria

Objective: The aim of this study is to detect in vitro the synergetic activity of colistin in combination with imipenem, amikacin or ciprofloxacin, at sub-inhibitory concentrations, against carbapenems-resistant (CR) Acinetobacter baumannii and Pseudomonas aeruginosa strains isolated from various wards in Annaba teaching hospital in eastern Algeria.
Materials and Methods: The minimal inhibitory concentrations (MIC) were determined by broth macrodilution (BMD). Carbapenemase encoding genes were screened using polymerase chain reaction (PCR). The activity of colistin in combination with second antibiotic was evaluated by the Checkerboard Technique.
Results: 39 CR P. aeruginosa and 21 CR A. baumanni strains where collected. The MIC values ranging from (0.25 to 4 µg/ml) to colistin, ≥16 µg/ml for imipenem, ≥4 µg/ml to amikacin and ≥8 µg/ml ciprofloxacin. The PCR reveals the presence of the genes bla_OXA23 (n = 12), blaOXA24 (n = 6), blaNDM1 (n = 3) in A. baumannii and blaVIM2 (n = 12) in P. aeruginosa. The combination of colistin with imipenem showed synergistic effect on 57.14% and 46.15% of A. baumannii and P. aeruginosa isolates, respectively. For colistin and amikacin, the synergistic effect is detected in 28.6% of A. baumannii and 30.8% of P. aeruginosa. While colistin and ciprofloxacin showed synergy on 14.29% and 15.38% of A. baumannii and P. aeruginosa isolates, respectively.
Conclusion: CR A. baumannii and P. aeruginosa remain the most prevalent infection agents in patients from high-risk wards at Annaba Hospital. Colistin associated with imipenem or with amikacin at sub-inhibitory concentrations gives very encouraging results allowing better management of infections caused by this type of bacteria.     

SPAG9 promotes prostate cancer growth and metastasis

Sperm-associated antigen 9 (SPAG9) expression is increased in prostate tissues of prostate cancer patients. This experimental study aimed to investigate the role of SPAG9 in bone metastasis of prostate cancer. Immunohistochemical analysis showed that SPAG9 staining was positive in 81.67% of 240 cases of prostatic carcinoma but only in 6.67% of 120 cases of benign prostate hyperplasia. Strong PAG9 staining was positively correlated with Gleason score and bone metastasis in 240 prostate cancer patients (p < 0.05), but not with the age or serum prostatespecific antigen level (p > 0.05). PC-3 cells were transfected with shRNA against SPAG9, and CCK-8 assay in triplicate showed that PC-3 cell viability was inhibited by SPAG9 knockdown. In addition, transwell assay in triplicate showed that PC-3 cell invasion was inhibited by SPAG9 knockdown. Furthermore, total 2 × 106 PC-3 cells were injected subcutaneously into the right flank of nude mice which were randomly divided into three groups (N = 8) and treated by intratumoral injection of SPAG9 shRNA, control shRNA or PBS, respectively. SPAG9 shRNA inhibited the growth, invasion and angiogenesis while promoted apoptosis of xenografted PC-3 cells. SPAG9 knockdown led to the upregulation of E-cadherin and the downregulation of MMP2 and vimentin in xenografted tumors. In conclusion, this is the first study to provide evidence that SPAG9 promotes bone metastasis of prostate cancer, and SPAG9 is a promising target to prevent or treat bone metastasis of prostate cancer.     

Calcitriol induces post-thawed bovine sperm capacitation

Background: Capacitation is a set of physiological changes sperms undergo to acquire fertilizing capacity. In vivo, this process is directly associated with high calcium levels in sperm cytoplasm. Calcitriol, the vitamin D hypercalcemic metabolite, is related to human sperm motility, capacitation, and acrosome reaction. This work aimed to study the effect of calcitriol on bull sperm quality parameters and capacitation. Methods: One million freeze-thawed spermatozoa were obtained from different bulls and treated with 20 nM of calcitriol for 30 min. Untreated cells (negative control) and treated ones with calcitriol or heparin (100 µg/mL, positive capacitation control) were evaluated for motility, viability, and functional parameters. Menadione (70 µM, 30 min) treatment was included as a reactive oxygen species (ROS) positive sperm agent. Results: The results elucidated that sperm exposed to 20 nM calcitriol showed higher viability, vigor, and capacitation than their positive and negative controls. The percentage of sperm with intact plasma and acrosome membranes, mitochondrial membrane potential (ΔΨm), and phosphatidylserine externalization was similar in all the conditions evaluated, while ROS production was higher with heparin and menadione-treated groups than the calcitriol group or negative control. Conclusion: Our results indicate that calcitriol induces the capacitation of thawed bull spermatozoa and maintains acceptable values of progressive motility, viability, and vigor without altering key biological parameters such as redox status, ΔΨm, and cell death.     

Micropropagation of <i>Ilex dumosa</i> (Aquifoliaceae) from nodal segments in a tissue culture system

Micropropagation of Ilex dumosa var. dumosa R. (“yerba señorita”) from nodal segments containing one axillary bud was investigated. Shoot regeneration from explants of six-year-old plants was readily achieved in ¹/₄ strength Murashige and Skoog medium (¹/₄ MS) plus 30 gr·L^-1 sucrose and supplemented with 4.4 µM BA. Further multiplication and elongation of the regenerated shoots were obtained by subculture in a fresh medium of similar composition with 1.5 gr·L^-1 sucrose. Rooting induction from shoots were achieved in two steps: 1) 7 days in ¹/₄ MS (30 gr·L^-1 sucrose, 0.25 % Phytagel^®) with 7.3 µM IBA and 2) 21 days in the same medium without IBA and 20 µM of cadaverine added. Regenerated plants were successfully transferred to soil. This micropropagation schedule can be implemented in breeding programs of Ilex dumosa.     

Edematogenic and myotoxic activities of the duvernoy’s gland secretion of <i>Philodryas olfersii</i> from the north-east region of Argentina

Philodryas olfersii is found in South America, from Amazonas to Patagonia. It is important to characterize the venom of P. olfersii, who inhabits the North-East region of Argentina, since snake venoms are known to exhibit considerable variability in composition and biological activities. In this work, mice weighing 18-20 g (n = 4 for each experimental group) were used. For the edematogenic activity mice were injected s.c. in the right foot pad with 50 µl of solutions containing different amounts of venom, whereas the left foot pad was injected with 50 µl of PBS. Two hours after injection mice were killed by cervical dislocation and both feet were cut off and weighed individually. For the myotoxic activity mice were injected i.m. with 100 µl of solutions containing 40 µg of venom. Blood samples were extracted after 1, 3, 6, 8, 10, 12, 14, 16 and 24 h of venom injection to determinate serum CPK activity and mice were sacrificed at the same time intervals to obtain the inoculated gastrocnemius muscle. They were fixed with Bouin solution and stained with Hematoxylin-Eosin. Results showed that P. olfersii venom exhibits a high edematogenic activity (MED = 0.31 µg) and a moderate myotoxic activity. Myonecrosis reached its highest level after 12 h of venom injection as shown by plasmatic CPK levels (5,401 ± 330 IU/l) and microscopic assay. It demonstrates the potential toxicity of the venom of P. olfersii, who inhabits the North-East region of Argentina. It also reinforces the original warning concerning the potential danger of bites by colubrids.     

Effects of exposure to estradiol and estradiol plus testosterone on the mongolian gerbil (Meriones unguiculatus) female prostate

The female prostate is a differentiated organ found in several mammal species, including humans and rodents. This gland has been related to important functions on female reproductive biology. Although the factors, which regulate prostate's development and activity are not well known, its functionality has been related to steroid hormones. It is well established that cyclic changes of estradiol and progesterone levels promote histophysiological adaptations of the whole female body. In contrast, only a few is found about those adaptations in female prostate. Thus, this study aimed to evaluate the effect of estradiol and estradiol + testosterone association on gerbil female prostate in order to verify, which hormonal associations are necessary to its homeostasis. For this, adult females had the ovaries surgically removed. After recovering, they received estradiol and estradiol + testosterone doses through 30 days, each 48 h. The prostatic tissue underwent morphological and morphometric‐estereological analysis. Hormonal restriction caused great gland involution and decreased secretory activity, aspects that were reverted by exposure to estradiol and estradiol + testosterone. However, these hormones were not able to re‐establish the normal prostate histoarchitecture. The immunoreaction of steroid receptors (ER‐α, ER‐β, and AR) responded differently among the experimental and control groups, and PCNA assay showed a decrease in epithelial cell proliferation within groups that had hormone privation. Therefore, we conclude that estradiol and testosterone are able to influence prostate morphophysiology and the maintenance of gland homeostasis depends on a balance among these and other hormones. Microsc. Res. Tech. 76:486–495, 2013. © 2013 Wiley Periodicals, Inc.     

Steroid hormone receptors, matrix metalloproteinases, insulin-like growth factor, and dystroglycans interactions in prostatic diseases in the elderly men

OBJECTIVES: The aim of this study was to evaluate the reactivity of steroid hormone receptors (SHRs), dystroglycans (DGs), matrix metalloproteinases (MMPs), insulin‐like growth factor receptor (IGFR‐1), and laminin (Lam) in both prostatic stromal and epithelial compartments showing different diseases in elderly men. METHODS: Sixty prostatic samples were obtained from 60‐ to 90‐year‐old patients (mean 63 years) with and without prostatic lesions from Hospital of the School of Medicine, State University of Campinas (UNICAMP). The Samples were divided into standard (no lesions); high grade prostatic intraepithelial neoplasia (HGPIN); prostatic cancer (PC); and benign prostatic hyperplasia (BPH) groups. The samples were submitted to immunohistochemistry and Western blotting analyses. Research Ethics Committee of the School of Medicine, University of Campinas/UNICAMP (number 0094.0.146.000‐08). RESULTS: The results showed increased IGFR‐1 and MMPs protein levels in the PC and HGPIN groups. Decreased αDG and βDG protein levels were verified in the PC and HGPIN groups. Androgen receptor (AR) reactivity was similar among all groups. Estrogen receptor α (Erα) immunoreactivity was more intense in the epithelium in the PC and HGPIN groups. Estrogen receptor β (ERβ) immunoreactivity was weak in the epithelium of the HGPIN and PC groups. CONCLUSIONS: To conclude, there was an association among IGFR‐1, MMPs, and SHRs, indicating IGFR‐1 as a target molecule in prostate therapy, considering the IGF proliferative properties. Also, the distinct SHRs reactivities in the lesions in both prostatic compartments indicated different paracrine signals and pointed out the importance of estrogenic pathways in the activation of these disorders. Microsc. Res. Tech. 75:1197–1205, 2012. © 2012 Wiley Periodicals, Inc.     

Increased androgen receptor and remodeling in the prostatic stroma after the inhibition of 5‐alpha reductase and aromatase in gerbil ventral prostate

Prostate require high levels of steroidogenic enzymes such as 5α‐reductase (5α‐r) and Aromatase (Aro) for the formation of active steroids. Dihydrotestosterone (DHT), the prostate dominant androgen, is converted from testosterone (T) by the action of 5α‐r. Aro provides an alternative pathway for estrogen, via T aromatization. Since prostatic maintenance is dependent on both reciprocal stromal–epithelial interaction and regulation by steroids, this study aimed to elucidate what the absence of 5α‐r and Aro enzymes provokes in the prostate microenvironment after their long‐term inhibition. Data obtained 1 day after the 30 consecutive days of enzymatic inhibition with Finasteride (5α‐r inhibitor) and Letrozole (Aro inhibitor) demonstrated a marked stromal remodeling, with an increased deposition of extracellular matrix (ECM) proteins besides androgen receptor (AR) overexpression in the three phases of postnatal development analyzed. The subepithelial area of acini from ventral prostate presented collagen and reticular fibers accumulation, besides various altered and active fibroblasts. The AR content immunostaining was elevated after enzymatic inhibition therapy, mainly in the nuclei of epithelial cells. Similar data were observed in the ventral prostates even 21 days after the end of treatments. Results obtained following the long‐term inhibition of 5α‐r and Aro are relevant and highlight the actions of these enzymes as crucial not only for the maintenance of tissue architecture and ECM arrangement but also for androgen and AR function. The long‐term absence of their action imposes a novel situation on the prostate from which its normal physiology could not be restored by the conclusion of the treatments. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

Elemental sulfur upregulated testicular testosterone biosynthesis by associating with altered gut microbiota in mice

Elemental sulfur has been used as a traditional Chinese medicine to treat the late-onset hypogonadism and impotence without a clarified mechanism for many hundreds of years. In the present study, mice were received sulfur or distilled water for 35 days by daily intragastric gavage at a dose of 250 mg/kg body weight. Then, the serum testosterone level and genes associated with testicular testosterone biosynthesis (TTB) were detected. The gut microbiota was also analyzed by 16S rRNA gene sequencing. Serum testosterone level was significantly increased by 291.1% in sulfur-treated mice. The H₂S levels in serum and feces were significantly increased. The expression of genes associated with TTB including StAR, p450c17, 3β-HSD, and P450scc in testes were significantly upregulated by Sulfur and NaHS, suggesting that sulfur promotes TTB depending on H₂S. In addition, sulfur increased the diversity of gut microbiota and the abundance of several bacteria associated with sulfur metabolism, including genus Prevotella, which might be positively associated with serum level of testosterone in boys. Five pathways including bile secretion, carotenoid biosynthesis, lipid biosynthesis proteins, propanoate metabolism, and biosynthesis of type II polyketide products, were identified to associate with sulfur. Together, our results suggested that sulfur upregulated testicular testosterone biosynthesis via H₂S, which was associated with alteration of gut microbiota in mice. Our study highlights a mechanism for the treatment of late-onset hypogonadism and impotence by sulfur.     

胶束电动色谱法测定辣椒中的辣椒碱

目的:建立分离辣椒碱和二氢辣椒碱,测定辣椒中辣椒碱的毛细管电泳新方法。方法:采用胶束电动毛细管色谱法,缓冲溶液体系由15mmol/L的硼砂、30 mmol/L的十二烷基硫酸钠组成,缓冲溶液的pH值为9.70,运行操作电压为20kV。结果:辣椒碱检测浓度在1～400mg/L范围内线性关系良好(r=0.9996),平均回收率为97%～103%(RSD<5%)。结论:该方法简单、快速、分析准确度高,能有效地分离辣椒碱和二氢辣椒碱,并测定辣椒中的辣椒碱。     

Doxazosin treatment alters stromal cell behavior and increases elastic system fibers deposition in rat prostate

Doxazosin (DOX), an α‐adrenoceptor antagonist, induces the relaxation of smooth muscle cell tonus and reduces the clinical symptoms of benign prostatic hyperplasia (BPH). However, the effects of DOX in the prostate stromal microenvironment are not fully known. In a previous study, we showed that DOX treatment for 30 days increased deposition of collagen fibers in the three rat prostatic lobes. Herein, we investigated the effects of DOX on stromal cell ultrastructure and elastic fiber deposition. Adult Wistar rats were treated with DOX (25 mg/kg/day); and the ventral, dorsal, and anterior prostates were excised at 30 days of treatment. The prostatic lobes were submitted to histochemical and stereological‐morphometric analyze and transmission electron microscopy (TEM). Histochemical staining plus stereological analysis of the elastic fiber system showed that DOX‐treated prostatic lobes presented more elaunin and elastic fibers than controls, mainly in the ventral lobe. Ultrastructural analysis showed that fibroblasts and smooth muscle cells from DOX‐treated prostates presented active synthetic phenotypes, evidenced by enlarged rough endoplasmic reticulum and Golgi apparatus cisterns, and confirmed the observation of thickened elaunin fibers. Our findings suggest that, under α‐adrenergic blockade by DOX, the fibroblasts become more active and smooth muscle cells shift from a predominantly contractile to a more synthetic phenotype. The deposition of collagen and elastic system fibers in the prostatic stroma may counterbalance the absence of smooth muscle tone during α‐blockers treatment. Microsc. Res. Tech. 73:1036–1044, 2010. © 2010 Wiley‐Liss, Inc.     

Role of foliar spray of plant growth regulators in improving photosynthetic pigments and metabolites in <i>Plantago ovata</i> (Psyllium) under salt stress–A field appraisal

Salinity is one of the major abiotic factors that limit the growth and productivity of plants. Foliar application of plant growth regulators (PGRs) may help plants ameliorate the negative impacts of salinity. Thus, a field experiment was conducted at the Botanical Garden University of Balochistan, Quetta, to explore the potential role of PGRs, i.e., moringa leaf extract (MLE; 10%), proline (PRO; 1 µM), salicylic acid (SA; 250 µM), and thiourea (TU; 10 mM) in ameliorating the impacts of salinity (120 mM) on Plantago ovata, an important medicinal plant. Salinity hampered plant photosynthetic pigments and metabolites but elevated oxidative parameters. However, foliar application of PGRs enhanced photosynthetic pigments, including Chl b (21.11%), carotenoids (57.87%) except Chl a, activated the defense mechanisms by restoring and enhancing the metabolites, i.e., soluble sugars (49.68%), soluble phenolics (33.34%), and proline (31.47%), significantly under salinity stress. Furthermore, foliar supplementation of PGRs under salt stress led to a decrease of about 43.02% and 43.27% in hydrogen peroxide and malondialdehyde content, respectively. Thus, PGRs can be recommended for improved photosynthetic efficiency and metabolite content that can help to get better yield under salt stress, with the best and most effective treatments being those of PRO and MLE to predominately ameliorate the harsh impacts of salinity.     

Isolation and biological characterization of a basic phospholipase A₂ from <i>Bothrops jararacussu</i> snake venom

A phospholipase A₂ has been isolated from Bothrops jararacussu venom from snakes that inhabit the northeast region of Argentina. The present study describes in vivo and in vitro biological activities of phospholipase A₂ from B. jararacussu as well as isolation details of its. Venom was obtained by milking of adult snakes which were housing in wood reptile cages of varying dimensions in heated (20–30ºC) rooms. Snakes received a weekly diet of mice and water was available ad libitum for drinking and soaking. The enzyme was purified by gel filtration on a Sephadex G-75 column followed by ion exchange chromatography on a SP-Sephadex C25 column. The major peak belonging to proteins was retained in the cation exchanger and then eluted using a concentration gradient of KCl that exhibited phospholipase activity.
This basic PLA₂ consists of a single polypeptide chain with a molecular mass of 15.6 kDa. It had a high indirect hemolytic activity and produced a significant paw edema reaction in mice. The enzyme showed a low lethality (LD₅₀ 148.6 µg) when was administered i.p. but exhibited elevated myotoxic effects in vivo by increasing plasma CK activity of injected mice, corroborated results by the histological observations of samples of gastrocnemius muscle. Myonecrosis is the result of intense destruction of muscular fibers that involves local infiltration of inflammatory cells and leads to the highest peak of CK level just after 1 hour mice injection. Moreover, the isolated enzyme showed anticoagulant activity, evaluated on sheep platelet-poor plasma which recalcification time was prolonged after incubation with the isolated phospholipase A₂ . These findings showed that this phospholipase, isolated by only two simple chromatographic steps, possesses high edematogenic and myotoxic activities. However, despite the low lethal activity, this enzyme would contribute markedly to the pathophysiology of the bothropic envenomation.     

Nitric oxide alleviates cadmium-impeded growth by limiting ROS accumulation in pea seedlings

Cadmium (Cd) causes oxidative stress, which leads to the oxidation of various biomolecules by the production of reactive oxygen species (ROS) to facilitate programmed cell death (PCD). The antioxidant defense system fails to detoxify ROS when it is produced in excess. Nitric oxide (NO), a gaseous free radical and a phytohormone, regulates various physiological processes of plants. Therefore, this work was undertaken to study the effects of the application of exogenous sodium nitroprusside (SNP, a NO donor) on growth parameters, oxidative stress, accumulation of secondary metabolites, and activities of antioxidant enzymes under Cd stress. Mild (50 µM) and severe (200 µM) Cd stress were applied to hydroponically grown pea (Pisum sativum L.) plants with or without 50 µM SNP. Severe Cd stress had a substantial impact on the plants. The effectiveness of NO in reducing Cd-induced negative effects on plant height, fresh weight, dry weight, protein content, nitrite content, nitrate reductase (NR) activity, catalase activity, and peroxidase activity were investigated. Seedling development, protein content, nitrite content, nitrate reductase (NR) activity, antioxidant defense systems disruption, overproduction of reactive oxygen species, and oxidative damage were observed. The antioxidant defense system (catalase and peroxidase activities) was activated by NO, which resulted in lower lipid peroxidation and lower hydrogen peroxide (H₂O₂) levels in Cd-exposed plants. SNP treatment boosted endogenous NO levels and NR activity in Cd-stressed plants while also enhanced proline levels to preserve osmotic equilibrium. The presence of total phenols and flavonoids increased after SNP treatment, indicating that SNP enhanced stress recovery and boosted plant development in Cd-stressed plants.

     

Lobe variation effects of experimental diabetes and insulin replacement on rat prostate

We investigated the impact of diabetes with simultaneous and late insulin replacement on rat prostate growth during puberty, paying special attention to different prostatic lobes. Diabetes was induced by administration of streptozotocin (STZ) in 40-day-old male Wistar rats. A subset of diabetic rats underwent simultaneous insulin replacement (3 days after STZ administration), and another subset underwent a late insulin replacement (20 days after STZ administration). The ventral, dorsolateral, and anterior prostatic lobes were weighed and processed for histological, immunohistochemical, and morphometric analyses. Both diabetic and insulin-treated animals maintained low plasma testosterone (T) concentrations, whereas dihydrotestostenore (DHT) levels were normal. Diabetic animals had a decreased gain in absolute prostatic weight when compared to age-matched controls and insulin replacement animals. However, prostatic lobe weight in the diabetic animals was ～100% higher, even at the beginning of the experiment. Among the lobes, the anterior lobe showed the highest weight gain in diabetic and insulin replacement conditions. Epithelial cell proliferation in all lobes was significantly reduced in diabetic animals and significantly increased in insulin replacement animals, although apoptosis was unaltered. In conclusion, diabetes diminishes, but does not abolish, prostate growth during puberty. Even late insulin administration reduces the adverse effects of this disease on the prostate. In a scenario with both low insulin and T levels, DHT and other factors may play an important role in pubertal prostate growth. The adverse effects of diabetes on the rat prostate show a variation in lobe response, suggesting that diabetes may affect human prostate zones differently.     

Comparative immunohistological study on using capsaicin,piperine, and okadaic acid for the transepithelial passage of the inactivated viral and bacterial vaccines in fish

The practical difficulty of parenteral application of fish vaccines against devastating fish diseases diverted the interest toward oral vaccination. Search for effective methods to enhance the oral uptake of viral and bacterial vaccines is continuing. The current research focus on a new role of mucosal fish vaccine adjuvants inducing the antigen uptake by enhancing vascularity or increasing intestinal permeability. Some inflammatory substances cause reversible pathology to the intestinal epithelium, which could be employed for the transepithelial passage of vaccine particles. The natural inflammatory substances used were capsaicin, piperine, and okadaic acid as 1 mg, 2 mg, and 1 μg/fish, respectively. Two inactivated vaccines were used as antigens to test the effect of these inflammatory substances in two different fish hosts. Tested vaccines were inactivated redspotted grouper nervous necrosis virus vaccine in sevenband grouper (Epinephelus septemfasciatus) and inactivated Edwardsiella tarda vaccine in red sea bream (Pagrus major) fish models. The inflammatory substances and each vaccine were anally intubated to fish. Capsaicin proved to be effectively aiding the transepithelial passage of vaccine particles more than piperine, while okadaic acid had no detectable effect.     

Curcumin gum Arabic nanoparticles demonstrate potent antioxidant and cytotoxic properties in human cancer cells

The main purpose of the study was to enhance the stability and therapeutic effects of Curcumin (Cur) through nanoformulation with gum Arabic (GA) as a coating agent through an efficient synthetic approach. The antioxidant properties of the developed nanoparticles (Cur/GANPs) were assessed through several in vitro assays, such as β-carotene bleaching activity, DPPH, and nitric oxide scavenging activities in addition to evaluating its inhibitory activity on angiotensin-converting enzyme (ACE). The cytotoxicity of Cur/GANPs was evaluated in vitro using different types of human cancer cells including breast cancer (MCF7, MDA-MB231), liver cancer (HepG2), and colon cancer (HT29) cells. The prepared particles displayed an elliptical shape with a size ranging between 20–260 nm and a potential difference of –15 mV. The Cur/GANPs exhibited significant antioxidant activity compared to free curcumin when using concentrations between 31.5 and 500 µg/mL. The Cur/GANPs also had inhibited the growth of all cancer cell lines in a proportional trend with concentrations used. Hence, the encapsulation with gum Arabic has augmented the antioxidant and anti-neoplastic effects of Curcumin. Therefore, Cur/GANPs may have effective therapeutic properties in diseases attributed to oxidative stress like cancer and hypertension.     

Insight into 5-aminolevulinic acid-induced modulation of cellular antioxidant metabolism to confer salinity and drought tolerance in maize

The current study investigated the comparative oxidative damage in two maize seedlings induced by saline, drought, and combined stress and the ameliorative role of two different doses (20 and 80 µM) of 5-aminolevulinic acid (ALA) against the above-mentioned stresses. Hydroponically grown 10-day-old maize (Zea mays, var. BARI Hybrid Maize-7 (BHM-7) and BARI Hybrid Maize-9 (BHM-9)) seedlings were exposed to 12 dS/m of saline solution, 200 mM mannitol-induced drought stress alone and their combined stress for 7 days. Result revealed that individual stresses retard the plant growth to some degrees; however, their combined stress has more detrimental effects, which might be correlated with lipid peroxidation (MDA)-induced oxidative stress in seedlings, enhanced Na⁺ /K⁺ ratio, and augmented generation of superoxide (O₂^•− ) and hydrogen peroxide (H₂O₂). In contrast, exogenous ALA supplementation at 20 µM concentration markedly recovered from chlorosis and growth inhibition, substantially scavenged reactive oxygen species (ROS) and MDA by preserving ionhomeostasis and relaxing oxidative stress; also, by boosting catalase (CAT) and glutathione S-transferase (GST), and exclusively via depressing the activity of lipoxygenase (LOX) antioxidant enzyme. On the contrary, 80 µM ALA made things worse; nevertheless, higher activities shown by other antioxidant enzymes; like, superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), and glutathione peroxidase (GPX), which were related to lessen the oxidative damage by highly produced O₂^•− and H₂O₂ under combined stress. Non-denaturing gel electrophoresis was done for further confirmation. However, ALA importantly increased the photosynthetic pigment contents in both genotypes irrespective of doses. Nevertheless, GST might have assisted the plants to escape from the herbicidal effect by detoxification. However, in the combined stress condition, high ALA concentration may have some positive role to play. Our findings also showed that BHM-9 performed better than BHM-7. Therefore, ALA at lower concentration was effective for single stress of saline and drought, while higher concentration can improve plant survival under combined stress.