Mitigation of water stress in broccoli by soil application of humic acid.

The main challenge to plant productivity is water scarcity, which is predicted to get worse with climate change, particularly in arid and semi-arid areas. Humic acid could improve plant tolerance to mitigate drought damage, which is an effective strategy to improve crop production and agriculture sustainability under limited water conditions in these regions, but its effective application rates should also be established. Thus, two field experiments were carried out at the Qaha Vegetable Research Farm in Qalubia Governorate, Egypt, during the two seasons of 2020-21 and 2021-22 on clay soil. The present study investigated the effect of three rates of humic acid application (0, 4.8, and 9.6 kg ha-1) on growth, yield, and quality of broccoli cv. Montop F1 hybrid under well-watered and drought conditions. Drought was induced by missing alternate irrigation. Soluble humic acid as potassium-humate was applied three times with irrigation water at the time of the first three irrigations of drought treatment. Water-stressed plants had a decrease in growth, yield, leaf chlorophyll, and nutrient content, while they showed an increase in the contents of leaf proline and curd dry matter and total soluble solids as well as water use efficiency, in both seasons. Soil application of humic acid was effective in mitigating the adverse effects of water deficit stress on the growth and yield of broccoli. Water-stressed plants had the highest WUE value (9.32 and 9.36 kg m3-1 in the first and second seasons, respectively) when the maximal humic acid rate was applied. Humic acid at a high level (9.6 kg ha-1) had the most promising results and represents an opportunity that must be applied to improve broccoli yield and its production sustainability in arid and semiarid regions.

Morphological, Biochemical, and Molecular Diversity Assessment of Egyptian Bottle Gourd Cultivars.

The genetic variability and relationships between ten bottle gourd cultivars were evaluated based on morphological, biochemical, and molecular parameters. The results displayed high variability among selected cultivars in terms of photosynthetic pigments, total free amino acids, total phenol content, isozymes pattern, and protein electrophoresis. Furthermore, differences in molecular markers were revealed by the SCoT technique. The peroxidase (POD) and polyphenyl oxidase (PPO) isozymes patterns did not detect significant differences in bands among cultivars. The protein patterns revealed seventeen bands ranging from 126 to 9 kDa and five polymorphic bands representing 29.41%. On the other hand, eight SCoT primers were used to evaluate the genetic variability and relationships between the ten Egyptian bottle gourd cultivars. The results of SCoT analysis detected 44 amplicons with 50% polymorphism. In addition, the results of the phylogenetic tree that is constructed based on the similarity coefficient revealed by SCoT analysis confirm the results of biochemical analysis indicating a genetic relationship between the most efficient bottle gourd cultivars (S1 and S2 cultivars). In addition, there is a genetic relationship among the less efficient bottle gourd cultivars (S4 and S5 cultivars). These results could be beneficial to distinguish among bottle gourd cultivars in the plant breeding programs.

Sorting lung tumor volumes from 4D-MRI data using an automatic tumor-based signal reduces stitching artifacts.

PURPOSE: To investigate whether a novel signal derived from tumor motion allows more precise sorting of 4D-magnetic resonance (4D-MR) image data than do signals based on normal anatomy, reducing levels of stitching artifacts within sorted lung tumor volumes. METHODS: (4D-MRI) scans were collected for 10 lung cancer patients using a 2D T2-weighted single-shot turbo spin echo sequence, obtaining 25 repeat frames per image slice. For each slice, a tumor-motion signal was generated using the first principal component of movement in the tumor neighborhood (TumorPC1). Signals were also generated from displacements of the diaphragm (DIA) and upper and lower chest wall (UCW/LCW) and from slice body area changes (BA). Pearson r coefficients of correlations between observed tumor movement and respiratory signals were determined. TumorPC1, DIA, and UCW signals were used to compile image stacks showing each patient's tumor volume in a respiratory phase. Unsorted image stacks were also built for comparison. For each image stack, the presence of stitching artifacts was assessed by measuring the roughness of the compiled tumor surface according to a roughness metric (Rg). Statistical differences in weighted means of Rg between any two signals were determined using an exact permutation test. RESULTS: The TumorPC1 signal was most strongly correlated with superior-inferior tumor motion, and had significantly higher Pearson r values (median 0.86) than those determined for correlations of UCW, LCW, and BA with superior-inferior tumor motion (p < 0.05). Weighted means of ratios of Rg values in TumorPC1 image stacks to those in unsorted, UCW, and DIA stacks were 0.67, 0.69, and 0.71, all significantly favoring TumorPC1 (p = 0.02-0.05). For other pairs of signals, weighted mean ratios did not differ significantly from one. CONCLUSION: Tumor volumes were smoother in 3D image stacks compiled using the first principal component of tumor motion than in stacks compiled with signals based on normal anatomy.

Effect of water stress and foliar application of chitosan and glycine betaine on lettuce.

The present study investigated the effect of foliar application of chitosan at 150 ppm and glycine betaine at 700 ppm on lettuce plants cv. Balady grown under well-watered and water deficit conditions in terms of growth, yield, quality, and water usage efficiency. The study was conducted in Qalubia Governorate, Egypt, during the two seasons of 2020/2021 and 2021/2022 on clay soil. Results indicated that water-stressed plants had a reduction in plant fresh weight, plant height, leaf area, and total yield, chlorophyll content and relative water content, while they exhibited an increase in total soluble solids, nitrate, and proline contents as well as water-use efficiency in both seasons. The foliar application of chitosan or glycine betaine to lettuce significantly improved plant performance under limited and normal irrigation conditions in comparison with untreated plants. The maximum positive effect was for chitosan foliar application. Overall, the results of this study indicated that foliar application of chitosan or glycine betaine was a substitute technology for improving the lettuce yield and quality as well as increasing water use efficiency under both irrigation regimes, but may be more efficient in lettuce plants subjected to a water deficit.

Effect of citric acid on phytoextraction potential of Cucurbita pepo, Lagenaria siceraria, and Raphanus sativus plants exposed to multi-metal stress.

Phytoextraction is a novel technique that involves using plants to remove heavy metals from contaminated soils. An outdoor pot experiment was designed to evaluate the phytoextraction potential of three plant species Cucurbita pepo, Lagenaria siceraria, and Raphanus sativus in soil contaminated with multiple metals (Cd, Co, Cr, Cu, Ni, Pb, and Zn) under the application of citric acid. The results showed that Raphanus sativus, out of all the studied plants, had the highest root and shoot dry weight and the capacity to accumulate all heavy metals at higher concentrations except for Cu. The application of citric acid into the polluted soil significantly increased plant growth, biomass, and heavy metal uptake. High bioconcentration values indicate that Raphanus sativus is a promising plant for absorbing and accumulating Cd and Ni from the soil. The maximum values of bioconcentration were also observed by the application of citric acid. The values of metal translocation from the root to the shoot were varied by plant species and the citric acid application. Regarding the biomass, metal content, as well as removal metal percentage values, it became apparent that the Raphanus sativus plant was the most effective crop in removing heavy metals from multi-metal contaminated Soil. Generally, these findings emphasize that the application of citric acid could be a useful approach to assist Cd and Ni phytoextraction by Raphanus sativus plants. When these plants are growing as vegetable crops, more attention should be given to evaluating the heavy metal content in them, especially when adding citric acid to their soil through fertigation systems to avoid food chain contamination.

Effects of biochar, zeolite and mycorrhiza inoculation on soil properties, heavy metal availability and cowpea growth in a multi-contaminated soil.

Heavy metal pollution of agricultural soil has become a major serious concern. The development of suitable control and remediation strategies for heavy metal contaminated soil has become critical. The outdoor pot experiment was conducted to investigate the effect of biochar, zeolite, and mycorrhiza on the bioavailability reduction of heavy metals and its subsequent effects on soil properties and bioaccumulation in plants as well as the growth of cowpea grown in highly polluted soil. Zeolite, biochar, mycorrhiza, zeolite with mycorrhiza, biochar with mycorrhiza, and soil without any modifications were the six treatments used. The experiment was conducted with a completely randomized design and four replications. The results indicated that the combination of biochar with mycorrhiza had the highest values of root and shoot dry weight and the lowest heavy metal concentrations in root and shoot as well as bioconcentration and translocation factors for all heavy metals. The highest significant reductions in the availability of heavy metals over the control were found with biochar with mycorrhiza, which were 59.1%, 44.3%, 38.0%, 69.7%, 77.8%, 77.2% and 73.6% for Cd, Co, Cr, Cu, Ni, Pb and Zn, respectively. The application of biochar and zeolite either alone or in combination with mycorrhiza increased significantly soil pH and EC compared to mycorrhiza treatment and untreated soil. It can be concluded that the combination of biochar and mycorrhizal inoculation has great potential as a cost-effective and environmentally technique for enhancing heavy metal immobilization, lowering heavy metal availability and plant uptake, and improving cowpea plant growth.

Cardamom oil ameliorates behavioral and neuropathological disorders in a rat model of depression induced by reserpine.

ETHNOPHARMACOLOGICAL RELEVANCE: Depression is a public health problem. Despite the availability of treatment options, its prevalence is increasing. A high rate of treatment failure is often reported, along with considerable side effects associated with synthetic antidepressants. Therefore, developing effective and safe antidepressants from traditional herbs or natural products as an alternative strategy is warranted to avoid side effects and increase drug efficacy. In traditional medicine, cardamom has traditionally been used to treat conditions like asthma, tooth and gum infections, cataracts, nausea, diarrhea, and even depression and anxiety as well as some problems with the heart, kidneys, and digestive system. AIM OF THE STUDY: The current study aimed to evaluate the antidepressant activity of cardamom oil in a rat model of depression induced by reserpine and compare it with the activity of the antidepressant drug fluoxetine. MATERIALS AND METHODS: Depression-like symptoms were induced in male rats by daily i. p. injection of reserpine (0.2 mg/kg/d for 15 d followed by 0.1 mg/kg/d for 21 d to maintain the depressive state), and the rats were treated with cardamom oil (oral dose = 200 mg/kg/d) for 21 d along with the maintenance dose of reserpine. We performed behavioral tests (forced swimming test and open-field test) and evaluated biochemical markers of depression. RESULTS: Our findings revealed that cardamom oil attenuated depression-like symptoms in reserpine-injected rats by improving the behavioral changes measured by the forced swimming test and the locomotor activities measured by the open-field test. In reserpine-injected rats, cardamom oil exerted antidepressant-like effects by modulating lower levels of brain monoamine neurotransmitters (serotonin, norepinephrine, and dopamine), GSH, and higher oxido-nitrosative stress parameters (malondialdehyde and nitric oxide). Moreover, cardamom oil alleviated depression-like behaviors by lowering monoamine oxidase activity and raising the activities of Na+/K+-ATPase and acetylcholinesterase and levels of a brain-derived neurotrophic factor in the cortex and hippocampus. CONCLUSION: We recommend the use of cardamom oil as a safe and reliable treatment or an adjuvant for preventing depression-like symptoms in patients suffering from depression.

Investigation of the effect of mutual vaccination with pest des petits ruminants and polyvalent foot and mouth disease vaccines on the immune response of sheep.

Background: Pest des petits ruminants (PPRs) and foot and mouth disease (FMD) are two viral infectious diseases affecting sheep dramatically causing great economic losses. Therefore, attention should be directed toward their control, especially through the application of well-designed vaccination schedules with specific potent vaccines. Aim: Determination of the possibility of sheep vaccination with PPR and FMD vaccines in a mutual schedule. Methods: Different groups of sheep have vaccinated with live attenuated PPR vaccine and inactivated polyvalent FMD vaccine in a mutual manner (one before the other at weekly intervals or simultaneously) followed by monitoring of the induced immunity to both vaccines using serum neutralization test (SNT) and enzyme linked immune sorbent assay (ELISA). Results: SNT and ELISA revealed that there was no antagonizing effect of any vaccine on the immune response to the mutual vaccination of sheep to the other where the obtained antibody titers in single vaccinated sheep groups were similar to those in the simultaneous vaccinated group. Conclusion: Simultaneous vaccination of sheep with PPR and polyvalent FMD vaccine is of applicable benefit saving time, effort, and stress factors on the animals.

Screening and assessing of pesticide residues and their health risks in vegetable field soils from the Eastern Nile Delta, Egypt.

The harmful effects of pesticide misuse on human health and the environment have become evident; so, this study aimed to monitor pesticide residues in soils of vegetable fields collected from the Eastern Nile Delta region, Egypt and to assess the potential health risks associated with them. Pesticide residues were determined using gas chromatography-mass spectrometry. Obtained results revealed that 100% of collected samples were contaminated with pesticides; residues of 33 compounds were detected in analyzed samples belonging to different chemical groups. Most detected pesticides (44%) were non-persistent and 40% were moderately persistent. While 1313% and 3% were persistent and very persistent compounds, respectively. Also, 36.7% and 30% of samples have two and three pesticides. Chlorpyrifos and propamocarb were the most dominant compounds that had widespread use across the study area. The number of detected pesticides per crop ranged from 1 to 16 (potato soil), followed by cucumber and tomato (13 pesticides), while one compound was detected in sweet potato soil. Soil organic matter content had a positive correlation with the total concentration of pesticide residues; however, no correlations were found with soil clay, pH and electrical conductivity contents. The human health risks of pesticides in the study soils were within acceptable levels. However, more attention should be paid in the future to decreasing the pesticide load and take place pesticide residue monitoring on vegetable soils.

Sensitivity and specificity of microRNA-204, CA125, and CA19.9 as biomarkers for diagnosis of ovarian cancer.

BACKGROUND: Ovarian cancer is usually detected at later stages and no effective screening approach, has been identified. Therefore, sensitive and specific biomarkers for detecting ovarian cancer are urgently needed. OBJECTIVE: This study aimed to investigate the efficacy of six biomarkers for the early clinical diagnosis of ovarian cancer. SUBJECTS & METHODS: The study included 120 patients (benign ovarian tumors and early and late ovarian carcinoma) and 30 control healthy volunteers. MiRNA-204, CA125, CA19.9, hepcidin, microfibril-associated glycoprotein 2, and ferroportin levels were determined in all patients and control volunteers. RESULTS: The combined area under the receiver operating characteristic curves for miRNA-204, CA125, and CA19.9 were 0.938, 1.000, and 0.998 for benign tumors and early and late ovarian carcinomas, respectively. The sensitivities of miRNA-204, CA125, and CA19.9 were 98.04%, 100.00%, and 96.19% and the specificities were 58.33%, 62.50%, and 57.78%, respectively. CONCLUSION: The positive predictivity of miRNA-204, CA125, and CA19.9 for ovarian cancer is high (59.57%, 58.24%, and 61.67%, respectively). Thus, the combination of these three biomarkers is a good diagnostic tool for ovarian cancer.

Pollution and health risk assessment of trace metal in vegetable field soils in the Eastern Nile Delta, Egypt.

The accumulation of trace metals in vegetable field soils is of increasing worry because of the potential health hazards and their detrimental effects on soil ecosystems. To investigate the state of trace metal pollution in vegetable field soils, 60 surface soil samples were collected from vegetable fields across the Eastern Nile Delta region, Egypt. The results concluded that the concentrations of Cu, Mn, and Ni were lesser than their corresponding background values, while the concentrations of Cd, Co, Pb, and Zn were exceeding their background values. The pollution indices showed that the studied soil experienced low to moderate contamination and the Cd and Cr contamination was serious. The hazard index values of nine trace metals signified that there was no adverse non-carcinogenic risk for adults and children. The carcinogenic risk of Cd, Co, Ni, and Pb for both age groups was within acceptable limits, while Cr had critical carcinogenic hazards for children. Overall, the quality of studied soils is relatively safe, although some samples impose serious pollution problems of Cd and Cr. Thus, properly monitored trace metals and soil management action should be applied to reduce further soil pollution in vegetable fields in the Eastern Nile Delta.

Active aspiration versus simple compression to remove residual gas from the abdominal cavity after laparoscopic cholecystectomy: a randomized clinical trial.

PURPOSE: After laparoscopic surgical procedures, residual gas in the abdominal cavity can cause post-operative pain, which is commonly located in the shoulder region. Previous studies suggested that post-laparoscopy pain can be prevented by active suctioning of intraabdominal gas at the end of surgery. METHODS: This randomized controlled trial (registered at DRKS 00,023,286) compared active suctioning versus manual compression in their ability to reduce pain after laparoscopic cholecystectomy. Patients scheduled for laparoscopic cholecystectomy were eligible for trial participation. The primary outcome measure was post-operative pain intensity after 12 h. All the patients were examined by MRI scanning to quantify the intraabdominal gas volume after the intervention. RESULTS: As planned, 60 patients were recruited. The two groups (n = 30 each) were very similar at the end of surgery. Active suctioning reduced the amount of residual pneumoperitoneum more than simple compression (median volume 1.5 versus 3.0 ml, p = 0.002). The primary outcome measure, abdominal pain after 12 h, was slightly lower in the intervention group (- 0.5 points, 95% confidence interval + 0.5 to - 1.7), but without reaching statistical significance (p = 0.37). After 12 h, shoulder pain was present in 10 patients in each group (p = 1.0). Independent of group assignment, however, residual gas volume was significantly associated with higher pain intensity. CONCLUSIONS: Active suctioning appears to have only a minor preventive effect on post-laparoscopy pain, probably because evacuation of the pneumoperitoneum remains incomplete in some patients. Other more effective maneuvers for gas removal should be preferred.

Antidepressant-like effects of aqueous extracts of miswak (Salvadora persica) and date palm (Phoenix dactylifera) on depression-like behaviors using CUMS model in male rats.

This study aimed to evaluate the antidepressant-like effects of aqueous extracts of miswak (Salvadora persica) and date palm (Phoenix dactylifera) and their combination on depression-like behaviors using a chronic unpredictable mild stress (CUMS) rat model of depression and to investigate the underlying possible mechanisms. Results showed that CUMS induced depression-like behaviors and anxiety in male rats, as determined by behavioral tests (FST, EPM, and OFT). CUMS significantly increased the levels of plasma malondialdehyde (MDA), cortisol, and pro-inflammatory cytokines (TNF-α and IL-6), in addition to enhancing acetylcholinesterase (ACHE) activity while plasma total antioxidant capacity and serotonin level were reduced. In the prefrontal cortex, CUMS decreased the expression of CREB and BDNF mRNA. However, aqueous extracts of miswak and date palm and their combination effectively ameliorated depressive-like behaviors, body weight loss, and oxidative stress induced by CUMS and restored serotonin and cortisol secretion to normal levels. Furthermore, the studied extracts improved the levels of plasma pro-inflammatory cytokines and CREB and BDNF mRNA expression in the prefrontal cortex. In conclusion, aqueous extracts of miswak and date palm have significant antidepressant-like effects on depression-like behaviors in CUMS model in rats. Moreover, their combination has higher antidepressant-like effects than either extract alone, suggesting that it may be an effective treatment for stress-induced depression-like behaviors acting through modulating pro-inflammatory cytokines and CREB/BDNF signaling pathway in the prefrontal cortex. PRACTICAL APPLICATIONS: Chronic stress is a major contributor to the development of depression. In recent years, there has been a significant increase in the use of herbal remedies for the treatment of a variety of diseases, particularly mood disorders. Because of the side effects encountered by antidepressant drugs such as anxiety, sexual dysfunction, loss of appetite, and inadequate response or developing tolerance to these medications, there is a need for more efficient and convenient antidepressant treatments. According to this study, aqueous extracts of miswak and date palm are effective treatments for stress-induced depression-like behaviors.

Effect of vas ligation on testicular sperm extraction results in nonobstructive azoospermic rats: Shaeer's lock-in technique.

This study evaluates the efficacy of vas ligation in enhancing sperm retrieval in nonobstructive azoospermia cases, by accumulating intratesticular spermatozoa. Fifty-six mature male rats with equally sized testes were included in this study. Forty-six were in the study group, and 10 were in the control group. Bilateral testicular fine needle aspiration was performed for all, to confirm presence of spermatozoa in all testes. Nonobstructive azoospermia was induced in all 56 rats, using Dienogest (40 mg/kg) + Testosterone Undecanoate (25 mg/kg) every month for three months. Monthly aspirations confirmed nonobstructive azoospermia from all rats, within the three months treatment. This was followed by unilateral vas ligation and was performed for 46 rats of the study group, with no ligation performed in the control group. After a further period of 90 days (2 spermatogenic cycles) with the same medical treatment maintained, bilateral testicular sperm extraction was performed. Sperm retrieval was evaluated, comparing the outcome of vas-ligated testicles to the nonligated. Upon evaluation, spermatozoa were found in 14/46 of the vas-ligated testes (30.4%), compared to none of the nonligated (0/66), p = .0005. Ligation of the vas deferens in rats with nonobstructive azoospermia may enhance the results of sperm retrieval via sperm accumulation.

Suppression of inflammatory cascades via novel cinnamic acid nanoparticles in acute hepatitis rat model.

Hepatitis was characterized by extreme inflammation and hepatocellular damage. Therefore, the current study aimed to gain insights into the modulation role of Cinnamic acid nanoparticles (CANPs) against acute hepatitis induced by d-Galactosamine and gamma radiation exposure (D-Gal/radiation) in the rat model and to suggest the implied molecular mechanism of CANPs. Acute hepatitis seriousness and the serum enzyme activities of ALT, AST, and ALP have been diminished upon oral administration of CANPs. Besides, the hepatic tissue levels of malondialdehyde (MDA) and nitric oxide (NO) have been significantly decreased, and the total antioxidant activity (TAO) depletion was extremely restored. Furthermore, the reduction of hepatic damage caused by pretreatment with CANPs was accompanied by significant suppression in the levels of hepatic proinflammatory cytokines (TNF-α, IL-1ß, and IL-18), NF-κB, NLRP3, caspase-1 and proapoptotic protein BAX whereas anti-apoptotic protein Bcl-2 level significantly elevated as compared with D-Gal/radiation-induced acute hepatitis (AH) group. Also, CANPs suppress the D-Gal/radiation-induced IL-1ß, IL-18, and ASK1 mRNA gene expression and the protein expression of TLR4 and MyD88 in the hepatic tissue. These biochemical parameters are confirmed by histological examination of the liver tissues. The present results indicated that CANPs can protect the hepatic cells from damage by both its anti-inflammatory and antioxidant influence as well as by modulating oxidation cellular pathways that have contributed to the acute severity of hepatitis. Also, CANPs is capable of suppressing apoptosis. Consequently, Nanoparticles of Cinnamic acid have the medicinal ability to protect the liver from acute hepatitis.

An audit of the management of elderly patients with glioblastoma in the UK: have recent trial results changed treatment?

Aim: We investigated uptake of short-course chemo-radiotherapy and compared outcomes with other treatment schedules in elderly patients with glioblastoma (GBM). Methods: Patients aged 65 or over with a diagnosis of GBM were identified from an 18-month period from three centers in the UK. The primary end point of this study was overall survival from the date of diagnosis. Results: The analysis included 210 patients. Overall median survival was 5.0 months. Approximately 31.9% of patients received combined chemoradiation; multivariate analysis showed that patients who received standard chemoradiation were at a reduced risk of death than those receiving hypofractionated chemoradiation. Discussion: In this retrospective study, patients treated with standard chemoradiation experienced better outcomes than patients receiving hypofractionated chemoradiation. Patient selection likely contributed to these findings.

Seed priming to alleviate salinity stress in germinating seeds.

Salinity is one of the major abiotic stresses that affect crop production in arid and semiarid areas. Seed germination and seedling growth are the stages most sensitive to salinity. Salt stress causes adverse physiological and biochemical changes in germinating seeds. It can affect the seed germination and stand establishment through osmotic stress, ion-specific effects and oxidative stress. The salinity delays or prevents the seed germination through various factors, such as a reduction in water availability, changes in the mobilization of stored reserves and affecting the structural organization of proteins. Various techniques can improve emergence and stand establishment under salt conditions. One of the most frequently utilized is seed priming. The process of seed priming involves prior exposure to an abiotic stress, making a seed more resistant to future exposure. Seed priming stimulates the pre-germination metabolic processes and makes the seed ready for radicle protrusion. It increases the antioxidant system activity and the repair of membranes. These changes promote seed vigor during germination and emergence under salinity stress. The aim of this paper is to review the recent literature on the response of plants to seed priming under salinity stress. The mechanism of the effect of salinity on seed germination is discussed and the seed priming process is summarized. Physiological, biochemical and molecular changes induced by priming that lead to seed enhancement are covered. Plants' responses to some priming agents under salinity stress are reported based on the best available data. For a great number of crops, little information exists and further research is needed.

Comparative study on the immunopotentiator effect of ISA 201, ISA 61, ISA 50, ISA 206 used in trivalent foot and mouth disease vaccine.

AIM: A comparison study was conducted to explore the best internationally available adjuvant that could be used in production of a highly potent foot and mouth disease (FMD) vaccine, that could stimulate a strong immune response and possibly give greater protection against FMD. MATERIALS AND METHODS: Four experimental batches of trivalent FMD vaccine were prepared with different available oil adjuvants which included Montanide ISA 201, 206, 61 and 50. RESULTS: The results indicated that vaccines emulsified using Montanide ISA 201 and Montanide ISA 206 adjuvants elicited a protective humoral immune response from the 2(nd) week postvaccination (WPV) as for ISA 201 with serum neutralization test (SNT) and enzyme-linked immune sorbent assay (ELISA) antibody titers of 1.62±0.047(a) and 1.8±0.049(a), 1.59±0.076(a) and 1.836±0.077(a), and 1.71±0.06(b) and 1.96±0.074(b) for serotypes O, A, SAT2, respectively, and for ISA 206 at SNT and ELISA antibody titers of 1.5±0.082(a) and 1.84±0.084(a), 1.56±0.037(a) and 1.818±0.052(a), and 1.5±0.106(a,b) and 1.81±0.104(a,b) for FMD virus serotypes O, A and SAT2, respectively. For ISA 61 and ISA 50, the protective antibody titer appeared in the 3(rd) WPV. In the ISA 61 FMD vaccine, SNT and ELISA titer were 1.59±0.076(a) and 1.9±0.094(a), 1.53±0.056(a) and 1.83±0.070(a), and 1.5±0.082(a) and 1.84±0.094(a) for serotypes O, A and SAT2, respectively, and in the case of ISA 50 FMD vaccine, the SNT, and ELISA titer were recorded for serotypes O, A and SAT2 respectively, 1.59±0.037(a) and 1.8±0.030(a), 1.68±0.056(a,b) and 1.916±0.065(a,b), and 1.65±0.082(a) and 1.9±0.09(a). On estimating the cellular immune response, the highest delta optical density levels for ISA 201 (0.395-0.460) and ISA 206 (0.375-0.428) were observed on 14 and 21 days post vaccination (DPV) respectively, while the highest levels of lymphoproliferation for ISA 61 (0.375-0.455) and ISA 50 (0.411-0.430) were on 21 and 28 DPV, respectively. CONCLUSION: The duration of immunity from Montanide ISA oils (201, 206, 61 and 50) FMD vaccines is a long-lived immunity which ranged between 32 and 38 weeks post vaccination but the Montanide ISA 201 FMD vaccine is superior to the others in the rapid cellular immune response of the vaccinated animals which showed its highest level within 14 days post vaccination.

Validation of γ-radiation and ultraviolet as a new inactivators for foot and mouth disease virus in comparison with the traditional methods.

AIM: The present work deals with different methods for foot and mouth disease virus (FMDV) inactivation for serotypes O/pan Asia, A/Iran05, and SAT-2/2012 by heat, gamma radiation, and ultraviolet (UV) in comparison with the traditional methods and their effects on the antigenicity of viruses for production of inactivated vaccines. MATERIALS AND METHODS: FMDV types O/pan Asia, A/Iran05, and SAT-2/2012 were propagated in baby hamster kidney 21 (BHK21) and titrated then divided into five parts; the first part inactivated with heat, the second part inactivated with gamma radiation, the third part inactivated with UV light, the fourth part inactivated with binary ethylamine, and the last part inactivated with combination of binary ethylamine and formaldehyde (BEI+FA). Evaluate the method of inactivation via inoculation in BHK21, inoculation in suckling baby mice and complement fixation test then formulate vaccine using different methods of inactivation then applying the quality control tests to evaluate each formulated vaccine. RESULTS: The effect of heat, gamma radiation, and UV on the ability of replication of FMDV "O/pan Asia, A/Iran05, and SAT-2/2012" was determined through BHK cell line passage. Each of the 9 virus aliquots titer 10(8) TCID50 (3 for each strain) were exposed to 37, 57, and 77°C for 15, 30, and 45 min. Similarly, another 15 aliquots (5 for each strain) contain 1 mm depth of the exposed samples in petri-dish was exposed to UV light (252.7 nm wavelength: One foot distance) for 15, 30, 45, 60, and 65 min. Different doses of gamma radiation (10, 20, 25, 30, 35, 40, 45, 50, 55, and 60 KGy) were applied in a dose rate 0.551 Gy/s for each strain and repeated 6 times for each dose. FMDV (O/pan Asia, A/Iran05, and SAT-2/2012) were inactivated when exposed to heat ≥57°C for 15 min. The UV inactivation of FMDV (O/pan Asia and SAT-2) was obtained within 60 min and 65 min for type A/Iran05. The ideal dose for inactivation of FMDV (O/pan Asia, A/Iran05, and SAT-2/2012) with gamma radiation were 55-60 and 45 kGy, respectively. Inactivation of FMDV with binary was 20, 24 and 16 hr for O/pan Asia, A/Iran05, and SAT-2/2012, respectively while inactivation by (BEI+FA) was determined after 18, 19 and 11 hr for O/pan-Asia, A/Iran 05, and SAT-2/2012, respectively. The antigenicity of control virus before inactivation was 1/32, it was not changed after inactivation in case of gamma radiation and (BEI+FA) and slightly decrease to 1/16 in case of binary and declined to 1/2, 1/4 in case of heat and UV inactivation, respectively. The immune response induced by inactivated FMD vaccines by gamma radiation and (BEI+FA) lasted to 9 months post-vaccination, while the binary only still up to 8 months post-vaccination but heat and UV-inactivated vaccines were not effective. CONCLUSION: Gamma radiation could be considered a good new inactivator inducing the same results of inactivated vaccine by binary with formaldehyde (BEI+FA).

The validity of testicular aspirate cytology and DNA image-analysis of the aspirate in the assessment of infertile men.

OBJECTIVE: To assess the possibility of using cytological examination and DNA image-analysis of testicular fine-needle aspirates instead of open surgical biopsy in the investigation of infertile men, as testicular biopsy has long been used for investigating infertility but the interpretation of histological slides is usually subjective. PATIENTS AND METHODS: Thirty-three men (aged 22-36 years) were evaluated for infertility and underwent both open biopsy and fine-needle aspiration of their testes. Subsequently, the needle aspirates were assessed histopathologically and cytologically, and by DNA image cytometry. The percentages of haploid, diploid and tetraploid cells were determined for each patient. RESULTS: The cases were divided into four categories: (1) Complete spermatogenesis, with a DNA pattern of 1n > 2n > 4n; (2) Maturation arrest, with a DNA pattern of 2n > 4n with no haploid cells; (3) Sertoli cell-only syndrome, with a DNA pattern of only 2n, with no haploid or tetraploid cells; (4) Hypospermatogenesis, with a variable DNA pattern, i.e. mild with 1n > 2n, moderate with 2n > 1n > 4n, and marked where the DNA pattern was 2n > 4n > 1n. From the cytological and DNA image-analysis of the aspirate a diagnosis was possible that had a strong correlation with the histological diagnosis of the same case. From image analysis we could exclude interstitial cells, Sertoli cells and sperms on the static image, and differentiate between spermatozoa and spermatids based on morphological characteristics in the cytological smear. This technique can therefore be used to quantitatively determine the percentages of various cell types within the seminiferous tubules. By coupling image ploidy analysis and cytological examination of a cytological smear, spermatogenesis can be assessed accurately. CONCLUSION: Image cytometry could be used to exclude interstitial cells, Sertoli cells and sperms on the static image and so produce an accurate assessment of spermatogenesis. A combination of ploidy and cell morphology characteristics in cytological smears provides an accurate, reproducible and easily used alternative to open testicular biopsy.