Copper Oxide Nanoparticles Induced Growth and Physio-Biochemical Changes in Maize (Zea mays L.) in Saline Soil.

Research on nanoparticles (NPs) is gaining great attention in modulating abiotic stress tolerance and improving crop productivity. Therefore, this investigation was carried out to evaluate the effects of copper oxide nanoparticles (CuO-NPs) on growth and biochemical characteristics in two maize hybrids (YH-5427 and FH-1046) grown under normal conditions or subjected to saline stress. A pot-culture experiment was carried out in the Botanical Research Area of "the University of Lahore", Lahore, Pakistan, in a completely randomized design. At two phenological stages, both maize hybrids were irrigated with the same amount of distilled water or NaCl solution (EC = 5 dS m-1) and subjected or not to foliar treatment with a suspension of CuO-NPs. The salt stress significantly reduced the photosynthetic parameters (photosynthetic rate, transpiration, stomatal conductance), while the sodium content in the shoot and root increased. The foliar spray with CuO-NPs improved the growth and photosynthetic attributes, along with the N, P, K, Ca, and Mg content in the roots and shoots. However, the maize hybrid YH-5427 responded better than the other hybrid to the saline stress when sprayed with CuO-NPs. Overall, the findings of the current investigation demonstrated that CuO-NPs can help to reduce the adverse effects of salinity stress on maize plants by improving growth and physio-biochemical attributes.

The Glycine soja cytochrome P450 gene GsCYP82C4 confers alkaline tolerance by promoting reactive oxygen species scavenging.

Recent studies have demonstrated the crucial role of Cytochrome P450 enzymes (CYPs) in the production of secondary metabolites, phytohormones and antioxidants in plants. However, their functional characterization specifically under alkaline stress remains elusive. CYP82C4 was the key gene screened from a family of wild soybean CYPs in our previous studies. The aim of this present study was to clone the Glycine soja GsCYP82C4 gene and characterize its functions in Arabidopsis and Glycine max. The results showed that the GsCYP82C4 gene displayed a high expression in different plant tissues at mature stages compared to young stages. Further, higher temporal expression of the GsCYP82C4 gene was noted at 6, 12 and 24 h time points after alkali treatment in leaves compared to roots. In addition, overexpression of GsCYP82C4 improved alkaline stress tolerance in Arabidopsis via increased root lengths and fresh biomass and strengthened the antioxidant defense system via a reduction in superoxide radicals in transgenic lines compared to wild type (WT) and atcyp82c4 mutants. Further, the expression levels of stress-related marker genes were up-regulated in GsCYP82C4 OX lines under alkali stress. The functional analysis of GsCYP82C4 overexpression in soybean displayed better hairy root growth, increased fresh weight, higher antioxidant enzyme activities and reduced lipid peroxidation rates in OX lines compared to the soybean WT (K599) line. In total, our study displayed positive roles of GsCYP82C4 overexpression in both Arabidopsis and Glycine max to alleviate alkaline stress via altering expression abundance of stress responsive genes, stronger roots, higher antioxidant enzyme activities as well as reduced rates of lipid peroxidation and superoxide radicals.

Uroprotective Potential of Campesterol in Cyclophosphamide Induced Interstitial Cystitis; Molecular Docking Studies.

Cyclophosphamide (CYP) is commonly used to treat cancer of the ovaries, breast, lymph, and blood system and produces interstitial cystitis (IC) via its urotoxic metabolite: i. e., acrolein. The present study was aimed to investigate the uroprotective effect of campesterol (a steroidal phytochemical) in cyclophosphamide induced IC. IC was induced by CYP (150 mg/kg, i. p.) in rats. The Enzyme linked immunosorbent assays for oxidative stress markers and Polymerase Chain Reaction (PCR) for inflammatory cytokines were carried out. The Tissue Organ Bath Technique was used for the evaluation of the spasmolytic effect of campesterol. Different pharmacological antagonists have been used to explore the mechanism of action of campesterol. Treatment with campesterol (70 mg/kg) reduced nociception (55 %), edema (67 %), hemorrhage (67 %), and protein leakage significantly (94 %). The antioxidant activity of campesterol was exhibited by a fall in MDA, NO, and an elevation in SOD, CAT, and GPX levels. Campesterol presented anti-inflammatory potential by decreasing IL-1, TNF-α, and TGF-ß expression levels. Histologically, it preserved urothelium from the deleterious effect of CYP. Campesterol showed a spasmolytic effect by reducing bladder overactivity that was dependent on muscarinic receptors, voltage-gated calcium and KATP channels, and cyclo-oxygenase pathways. In silico studies confirmed the biochemical findings. The findings suggest that campesterol could be valorized as a possible therapeutic agent against cyclophosphamide-induced interstitial cystitis.

Strigolactone signaling gene from soybean GmMAX2a enhances the drought and salt-alkaline resistance in Arabidopsis via regulating transcriptional profiles of stress-related genes.

Strigolactone (SL) is a new plant hormone, which not only plays an important role in stimulating seed germination, plant branching, and regulating root development, but also plays an important role in the response of plants to abiotic stresses. In this study, the full-length cDNA of a soybean SL signal transduction gene (GmMAX2a) was isolated, cloned and revealed an important role in abiotic stress responses. Tissue-specific expression analysis by qRT-PCR indicated that GmMAX2a was expressed in all tissues of soybean, but highest expression was detected in seedling stems. Moreover, upregulation of GmMAX2a transcript expression under salt, alkali, and drought conditions were noted at different time points in soybean leaves compared to roots. Additionally, histochemical GUS staining studies revealed the deep staining in PGmMAX2a: GUS transgenic lines compared to WT indicating active involvement of GmMAX2a promoter region to stress responses. To further investigate the function of GmMAX2a gene in transgenic Arabidopsis, Petri-plate experiments were performed and GmMAX2a OX lines appeared with longer roots and improved fresh biomass compared to WT plants to NaCl, NaHCO3, and mannitol supplementation. Furthermore, the expression of several stress-related genes such as RD29B, SOS1, NXH1, AtRD22, KIN1, COR15A, RD29A, COR47, H+-APase, NADP-ME, NCED3, and P5CS were significantly high in GmMAX2a OX plants after stress treatment compared to WT plants. In conclusion, GmMAX2a improves soybean tolerance towards abiotic stresses (salt, alkali, and drought). Hence, GmMAX2a can be considered a candidate gene for transgenic breeding against various abiotic stresses in plants.

The Effect of Calcium Hydroxide, Triple Antibiotic Paste and Chlorhexidine on Pain in Teeth with Symptomatic Apical Periodontitis: A Randomised Controlled Trial.

BACKGROUND: One of the main reasons for post endodontic pain is the polymicrobial intracanal flora that may survive the initial disinfection. A single antimicrobial agent might not provide adequate disinfection, and an antimicrobial combination such as triple antibiotic paste was tested to achieve this goal. AIM: The study aimed to assess the efficacy of three intra-canal medicaments on post root canal preparation pain. MATERIALS AND METHODS: Eighty patients with single-rooted necrotic teeth and symptomatic apical periodontitis were randomly assigned to four treatment groups (n = 20). Their preoperative pain was recorded on Wong-Baker's FACES pain rating scale. After chemo-mechanical canal preparation, intracanal medications were applied to the groups (Group 1 (20% calcium hydroxide), Group 2 (2% chlorhexidine), Group 3 (tri-antibiotic paste), Group 4 (no medication (Control group)). Patients were instructed to record pain on the Wong-Baker FACES pain rating scale at 4, 48, 72 and 96 hrs, postoperatively. Pain scores were analyzed using a one-way ANOVA test and Tukey's post hoc test and, if significant, Dunn's test was used for pair-wise comparisons. The significance level was set at a p-value ≤ 0.05. RESULTS: Tukey's post hoc test revealed significantly lower pain scores in Group 3 compared with other groups at all follow up intervals. Dunnett's test showed significantly lower pain in Group 3 compared with the Control group at 48, 72 and 96 hrs, postoperatively. CONCLUSIONS: Triple-antibiotic paste showed effective pain control as an intracanal medication on necrotic teeth with symptomatic apical periodontitis.

Cloning and in silico characterization of an abiotic stress-inducible U-box domain-containing protein gene GsPUB8 from Glycine soja.

The ubiquitination pathway is involved in the posttranslational modification of cellular proteins. However, the role of E3 ubiquitin ligase family proteins under abiotic stress conditions remains unclear, particularly in soybean. The core objective of the current study was to isolate and functionally characterize the GsPUB8 protein gene from wild soybean (Glycine soja) by using a homologous cloning method to investigate its abiotic stress responses. The GsPUB8 is a 40,562 Da molecular weight protein with 373 amino acid residues. The sequence alignment revealed the presence of U-box domain while the phylogenetic analysis showed an abundance of PUB8 proteins in both monocot and dicot plants. Analysis of gene structure predicted the absence of introns along with the presence of one exon. Furthermore, the activity of the GsPUB8 protein was anticipated in the plasma membrane and its expression was persuaded with NaCl, ABA, PEG6000, and NaHCO3 treatments with considerably higher manifestation in roots than leaves although, expressed in both vegetative and reproductive parts of G. soja. GsPUB8 protein showed 54% and 32% sequence identity to U-box domain containing 8 and 12 proteins from Arabidopsis thaliana and Oryza sativa subsp. japonica, respectively. GsPUB8 exhibited relatively higher expression under saline and drought stress particularly in roots. Whereas, the 3D model of GsPUB8 protein was generated using the SWISS-MODEL. This study can be used to manipulate the GsPUB8 protein or GsPUB8 gene for transformation purposes and its functional characterization under abiotic stress conditions.

Effect of foliar application of nano-nutrients solution on growth and biochemical attributes of tomato (Solanum lycopersicum) under drought stress.

Introduction: Drought stress has drastically hampered the growth and yield of many crops. Therefore, environmentally safe agricultural techniques are needed to mitigate drought stress impact. To this end, foliar spray of nano-nutrients solution to (NNS) alleviate harmful aspects of drought stress. Methods: In a completely randomized design (CRD) experiment, seedlings were transplanted into pots at 2-3 leaf stage, each filled with loam-compost- organic manure soil (3:1:1). Plants were divided into two groups. (a) control group (b) applied stress group. Plants at vegetative stage were treated with 100% FC for control group and 60% FC for drought group, and these levels were maintained until harvesting. Three treatments of NNS with four levels i.e., 0%, 1%, 3% and 5% were given to all the pots after two weeks of drought stress treatment with a gap of 5 days at vegetative stage. Results and discussion: Application of 1% of nano-nutrient solution displayed an improvement in shoot length, shoot fresh and dry weight, number of leaves and flowers. Leaf chlorophylls and carotenoids and total phenolics contents were found maximum while minimum electrolyte leakage was observed at 3% application compared to control. Further, 1% application of NNS increased the Leaf RWC%, total soluble sugars, flavonoids contents. 5% NNS application exhibited higher total free amino acids with minimum lipid peroxidation rate in leaves of tomato under drought. Antioxidant enzyme activities increased in a concentration dependent manner as gradual increase was observed at 1%, 3% and 5%, respectively. Overall, this study introduced a new insights on using nano-nutrient solutions to maintain natural resources and ensure agricultural sustainability.

Effectiveness of Direct Observation of Procedural Skills (DOPS) in Postgraduate Training in Urology at Institute of Kidney Diseases, Peshawar.

OBJECTIVE: To determine the effectiveness of direct observation of procedural skills (DOPS) in the assessment and improving operative skills of postgraduate residents in urology. STUDY DESIGN: Comparative study. PLACE AND DURATION OF STUDY: Department of Urology, Institute of Kidney Diseases (IKD), Hayatabad Medical Complex, Peshawar, from March till December 2016. METHODOLOGY: Twenty postgraduate residents in urology were assessed for three commonly performed procedures in urology with structured proforma, specially designed for DOPS, using Likert scale from 1 to 5. Formative feedback was given to all the residents highlighting their strengths and weaknesses. During second phase, all the three procedures were performed by four consultants respectively with residents as learner in the second phase of DOPS. During the third phase, a repeat assessment of e-residents was performed to study the overall improvement in operative skills, respectively. RESULTS: In pre-DOPS assessment, only 7 (35%) out of 20 residents, passed in skill of cystoscopy, 6 (30%) residents passed in transurethral resection of prostate (TURP), while 3 (15%) residents passed in skills of uretero-renoscopy (URS). The mean score of overall ability to perform procedure during first phase of DOPS were 2.7 ±1.3 for cystoscopy, 2.1 ±0.97 for TURP, and 2.5 ±1.1 for ureterorenoscopy. Significant improvement (p 0.04) in passing was observed in third phase with 17 (85%), 14 (70%), and 14 (70%) residents were declared passed in cystoscopy, TURP and URS, respectively. The mean score of overall ability to perform procedure during the third phase was 4.7 ±0.7 for cystoscopy, 4.5 ±0.94 for TURP, and 4.7 ±0.94 for ureteroscopy. CONCLUSION: DOPS is effective tool for assessing and improving the skills of postgraduate residents in urology.

Temperature on admission among cases of neonatal sepsis and its association with mortality.

OBJECTIVE: To ascertain the prevalence of hyperthermia and different categories of hypothermia among the cases of probable and culture-proven neonatal sepsis, and to evaluate association of mortality rate with axillary temperature at admission. METHODS: This analytical study was conducted at Fazl-e-Omar Hospital, Rabwah, Pakistan, from January to December 2013, and comprised all cases of culture-proven and probable neonatal sepsis admitted in the neonatal intensive-care unit of the hospital. Using World Health Organisation classification, cases were categorised according to their temperature on admission, and outcome was recorded. SPSS 20 was used for data analysis. RESULTS: Of the 374 patients, 47(12.6%) died. Besides, 217(58%) cases had normal temperature, 49(13.1%) had hyperthermia, 50(13.4%) had mild hypothermia, and 58(15.5%) had moderate hypothermia. Mortality rate was high among cases with mild and moderate hypothermia i.e. 15(32.6%) and 15(33%), compared with the mortality rate in cases with normal temperature i.e. 12(6.1%). Mortality rate among cases with hyperthermia was 5(11.6%). In early onset cases, 33(16.2%) had mild and 45(22.1%) had moderate hypothermia. In late onset cases, 37(21.8%) had hyperthermia. CONCLUSIONS: Most cases with neonatal sepsis had normal temperature. Mild and moderate hypothermia were found to be associated with higher mortality rate.