Polyaniline/ZnO Hybrid Nanocomposite: Morphology, Spectroscopy and Optimization of ZnO Concentration for Photovoltaic Applications.

The appropriate combination of semiconducting polymer-inorganic nanocomposites can enhance the existing performance of polymers-only-based photovoltaic devices. Hence, polyaniline (PANI)/zinc oxide (ZnO) nanocomposites were prepared by combining ZnO nanoparticles with PANI in four distinct ratios to optimize their photovoltaic performance. Using a simple coating method, PANI, ZnO, and its nanocomposite, with varying weight percent (wt%) concentrations of ZnO nanoparticles, i.e., (1 wt%, 2 wt%, 3 wt%, and 4 wt%), were fabricated and utilized as an active layer to evaluate the potential for the high-power conversion efficiency of various concentrations, respectively. PANI/ZnO nanocomposites are characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), atomic force microscopy (AFM), Fourier transform infrared (FTIR) spectroscopy, ultraviolet-visible (UV-vis) absorption, energy dispersive X-ray (EDX), and I-V measurement techniques. The XRD analysis showed a distinct, narrow peak, which corresponds to the wurtzite ZnO (101) plane. The SEM analysis verified the production of the PANI/ZnO composite by demonstrating that the crystalline ZnO was integrated into the PANI matrix. The elemental composition was determined by energy dispersive X-ray analysis (EDX), which confirmed the existence of PANI and ZnO without any impurities, respectively. Using Fourier transform infrared (FTIR) spectroscopy, various chemical bonds and stretching vibrations were analyzed and assigned to different peaks. The bandgap narrowing with an increasing PANI/ZnO composition led to exceptional optical improvement. The I-V characterization was utilized to investigate the impact of the nanocomposite on the electrical properties of the PANI/ZnO, and various concentrations of ZnO (1 wt%, 2 wt%, 3 wt%, and 4 wt%) in the PANI matrix were analyzed under both light and dark conditions at an STC of 1.5 AM globally. A high PCE of 4.48% was achieved for the PANI/ZnO (3 wt%), which revealed that the conductivity of the PANI/ZnO nanocomposite thin films improved with the increasing nanocomposite concentration.

Neuroprotective effect of sumatriptan in pentylenetetrazole-induced seizure is mediated through N-methyl-D-aspartate/nitric oxide and cAMP response element-binding protein signaling pathway.

Seizure occurs as a result of uncontrolled electrical disturbances within the brain. Various biomolecules such as N-methyl-D-aspartate (NMDA), nitric oxide (NO), and cAMP response element-binding protein (CREB) have been implicated in the pathophysiology of seizure. Sumatriptan is a specific 5-Hydroxytryptamine 1B/1D receptor agonist and has neuroprotective effects in various neuropsychiatric disorders. In the current study, we tried to investigate the possible interaction of sumatriptan with NMDA/NO and CREB signaling pathway in PTZ induced seizure. For this purpose, various agonist and antagonist of NMDA such as MK-801 and Ketamine, NO precursor L-ARG, and NOS inhibitors L-NAME and 7-NI were co-administered with sumatriptan in PTZ induced seizure model. The level of nitrite in mice hippocampus was determined by Griess reaction. The gene expression of NR1, NR2A, NR2B, and CREB were quantified by quantitative real time-polymerase chain reaction (qRT-PCR). Furthermore, the involved neuronal nitric oxide synthase (nNOS) protein expression was examined via western blot analysis. Effective dose of sumatriptan (1.2 mg/kg) alone and subeffective dose of sumatriptan (0.3 mg/kg) in combination with NMDA and/or NO antagonist showed significant (P < 0.001) anticonvulsant activity in mice. Furthermore, sumatriptan significantly inhibited the PTZ-induced mRNA expression of NR2A (P < 0.0001), NR2B (P < 0.05), and CREB (P < 0.01). Also, the expression of nNOS protein in PTZ treated group was reversed by sumatriptan (P < 0.01). Hence, current findings suggest that the anticonvulsant effect of sumatriptan was due to down regulation of NMDA/NO and CREB signaling pathway.

Energizing periodic mesoporous organosilica (PMOS) with bismuth and cerium for photo-degrading methylene blue and methyl orange in water.

This work reported an efficient catalyst to reduce the organic pollutants by using an energetic periodic mesoporous organosilica (PMOS) supported with bismuth (Bi-PMOS) and cerium (Ce-PMOS). PMOS support was designed through co-condensation of sodium silicate and 3-methacryloxypropyltrimethoxysilane on polysorbate templates. The resultant PMOSs were fabricated with bismuth and cerium oxides to formulate Bi-PMOS and Ce-PMOS, respectively. These materials showed photo-degradations of methylene blue (MB, 74.7% and 41.1% with Bi-PMOS and Ce-PMOS, respectively) and methyl orange (MO, 53.2% and 39.4% with Bi-PMOS and Ce-PMOS, respectively). Such efficient photo-degradations were attributed to the precise doping of metallic nodes of Bi2 O3 and CeO2 on the porous structure of PMOS with high surface area. The results also showed that Bi and Ce were more effective in PMOS support for photo-degradation of dyes as the support provides more lifetime to photo-generated electron-hole pairs than other materials. Moreover, active reusability and high degradation efficiencies of Bi-PMOS and Ce-PMOS proved them better analytical tools to reduce organic pollutants under visible lights. PRACTITIONER POINTS: The oxides of bismuth and cerium have impressive photocatalytic characteristics. New material energizing mesoporous organosilica with bismuth and cerium for photo-degradation of methylene blue and methyl orange in water. The use of an efficient catalyst to reduce the organic pollutants by using an energetic periodic mesoporous organosilica (PMOS) supported with bismuth (Bi-PMOS) and cerium (Ce-PMOS).

In vitro Characterization and Release Studies of Combined Nonionic Surfactant-Based Vesicles for the Prolonged Delivery of an Immunosuppressant Model Drug.

BACKGROUND: Cyclosporine A (CsA) is an exceptional immunosuppressant used for the treatment of immune disorders. Niosomal vesicles are promising drug carriers that are formed by self-association of nonionic surfactants and cholesterol in an aqueous phase. The objective of the study was to formulate combined nonionic surfactant based vesicles and to evaluate their in vitro characterization, release studies and in vivo studies. MATERIALS AND METHODS: Five niosomal formulations (F7 to F11) were prepared using the thin film hydration method. The molar ratio of cholesterol and non-ionic surfactant taken was 1:1. In formulation F10, the combination of surfactants Span 20 and Brij 35 was used. The niosomes were characterized by zeta sizer and SEM for particle size analysis, in vitro drug release and stability studies. The pharmacokinetic studies were conducted on healthy albino rabbits. RESULTS: The size of niosome was found in the range of 427.1 nm to 972.3 nm. SEM image of optimized formulations F10 exhibit the spherical nature of niosomal vesicles. DSC thermograms of niosomal formulations exhibited a broadened endothermic peak. The stability study exhibited that all formulations are stable and negligible change of vesicle size and entrapment was observed with time. The percentage drug release was significantly higher as compared to CsA plain dispersion for all niosomal formulations at pH 1.2 and 7.4. The release kinetic behavior showed that all preparations were best described by zero order and can release active ingredient in a sustained manner. The pharmacokinetic data showed the test formulation (F10) possessed greater bioavailability as compared to the reference formulation (CsA aqueous dispersion). CONCLUSION: The formulation F10 demonstrated a comparatively more delayed rate of release with enhanced dissolution as compared to a single surfactant scheme. The F10 formulation can be a remarkable nanotechnology for prolonged delivery of CsA orally with improved dissolution profile and bioavailability.

Investigation on Hamburg Wheel-Tracking Device Stripping Performance Properties of Recycled Hot-Mix Asphalt Mixtures.

Moisture damage in hot mix asphalt pavements is a periodic but persistent problem nowadays, even though laboratory testing is performed to identify different moisture-susceptible mixtures. In this study, a Hamburg Wheel Tracking device (HWTD) was used for rutting tests which were conducted on control and a high percentage of recycled asphalt pavement (RAP), i.e., 30%, 50% and 100% of virgin mixtures, under air dry and water-immersed conditions. Similarly, the extracted bitumen from RAP was tested for binder physical properties. Results showed that the asphalt mixtures containing RAP have less rut depth as compared to the control mix both in air dry and immersion conditions and hence showed better anti-rutting properties and moisture stability. Stripping performance of control and RAP containing mixtures was also checked, concluding that the RAP mixture was greatly dependent on the interaction between the binder (virgin plus aged) and aggregates.

Involvement of NMDA receptors in the antidepressant-like effect of tramadol in the mouse forced swimming test.

Tramadol is an analgesic agent that is mainly used to treat moderate to severe pain. There is evidence that tramadol may have antidepressant property. However, the mechanisms underlying the antidepressant effects of tramadol have not been elucidated yet. Considering that fact that N-methyl-d-aspartate (NMDA) receptor signaling may play an important role in the pathophysiology of depression, the aim of the present study was to investigate the role of NMDA receptor signaling in the possible antidepressant-like effects of tramadol in the mouse forced swimming test (mFST). We found that tramadol exerted antidepressant-like effects at high dose (40mg/kg, intraperitoneally [i.p.]) in the mFST. Co-administration of non-effective doses of NMDA receptor antagonists (ketamine [1mg/kg, i.p.], MK-801 [0.05mg/kg, i.p.], or magnesium sulfate [10mg/kg, i.p.]) with sub-effective dose of tramadol (20mg/kg, i.p.) exerted significant antidepressant-like effects in the mFST. The antidepressant-like effects of tramadol (40mg/kg) was also inhibited by pre-treatment with non-effective dose of the NMDA receptor agonist NMDA (75mg/kg, i.p.). Our data suggest a role for NMDA receptor signaling in the antidepressant-like effects of tramadol in the mFST.

Homozygosity Mapping and Targeted Sanger Sequencing Identifies Three Novel CRB1 (Crumbs homologue 1) Mutations in Iranian Retinal Degeneration Families

Background: Inherited retinal diseases (IRDs) are a group of genetic disorders with high degrees of clinical, genetic and allelic heterogeneity. IRDs generally show progressive retinal cell death resulting in gradual vision loss. IRDs constitute a broad spectrum of disorders including retinitis pigmentosa and Leber congenital amaurosis. In this study, we performed genotyping studies to identify the underlying mutations in three Iranian families. Method: Having employed homozygosity mapping and Sanger sequencing, we identified the underlying mutations in the crumbs homologue 1 gene. The CRB1 protein is a part of a macromolecular complex with a vital role in retinal cell polarity, morphogenesis, and maintenance. Results: We identified a novel homozygous variant (c.1053_1061del; p.Gly352_Cys354del) in one family, a combination of a novel (c.2086T>C; p.Cys696Arg) and a known variant (c.2234C>T, p.Thr745Met) in another family and a homozygous novel variant (c.3090T>A; p.Asn1030Lys) in a third family. Conclusion: This study shows that mutations in CRB1 are relatively common in Iranian non-syndromic IRD patients.

Diagnosis of Splenic Lymphoma by Endoscopic Ultrasound Guided Fine Needle Aspiration: A Case Report and Review of the Literature.

Introduction. Splenic tumor is usually found as an incidental finding on CT of abdomen. Traditionally, ultrasound (US) or computed tomography (CT) guided biopsies were employed for the purpose of sampling; however they have been reported to have a complication rate of 5.3%. Endoscopic ultrasound-fine needle aspiration (EUS-FNA) has been recently utilized for the purpose of sampling splenic tumors. In literature there are 7 reported instances where splenic lymphoma was diagnosed using EUS-FNA. We present a case of follicular B cell lymphoma of the spleen diagnosed using EUS-FNA. Case Report. 58-year-old female presented to her primary care physician for left upper quadrant abdominal pain for one week. Physical exam was significant for left upper quadrant tenderness. Her laboratory tests were within normal limits. She underwent CT scan of abdomen which revealed approximately 5 cm × 5 cm mass in spleen. EUS-FNA of the spleen revealed a large hypoechoic, heterogeneous, well-demarcated mass measuring 54.7 mm × 43.0 mm. Fine needle aspiration was performed, and the sample was submitted for cytology and flow cytometry. Flow cytometry revealed a lambda monotypic population of B cells displaying dim CD19 and CD10. Diagnosis of B cell non-Hodgkin low grade follicular lymphoma was made. Conclusion. Endoscopic ultrasound with fine needle aspiration is a very rare but safe, reliable method of diagnosis of splenic lymphomas.

Agmatine enhances the antidepressant-like effect of lithium in mouse forced swimming test through NMDA pathway.

Depression is one the world leading global burdens leading to various comorbidities. Lithium as a mainstay in the treatment of depression is still considered gold standard treatment. Similar to lithium another agent agmatine has also central protective role against depression. Since, both agmatine and lithium modulate various effects through interaction with NMDA receptor, therefore, in current study we aimed to investigate the synergistic antidepressant-like effect of agmatine with lithium in mouse force swimming test. Also to know whether if such effect is due to interaction with NMDA receptor. In our present study we found that when potent dose of lithium (30mg/kg) was administered, it significantly decreased the immobility time. Also, when subeffective dose of agmatine (0.01mg/kg) was coadministered with subeffective dose of lithium (3mg/kg), it potentiated the antidepressant-like effect of subeffective dose of lithium. For the involvement of NMDA receptor in such effect, we administered NMDA receptor antagonist MK-801 (0.05mg/kg) with a combination of subeffective dose of lithium (3mg/kg) and agmatine (0.001mg/kg). A significant antidepressant-like effect was observed. Furthermore, when subeffective dose (50 and 75mg/kg) of NMDA was given it inhibited the synergistic effect of agmatine (0.01mg/kg) with lithium (3mg/kg). Hence, our finding demonstrate that agmatine have synergistic effect with lithium which is mediated by NMDA receptor pathway.

Atorvastatin attenuates the antinociceptive tolerance of morphine via nitric oxide dependent pathway in male mice.

The development of morphine-induced antinociceptive tolerance limits its therapeutic efficacy in pain management. Atorvastatin, or competitive inhibitor of 3-hydroxy-methyl-glutaryl coenzyme A (HMG-CoA) reductase, is mainstay agent in hypercholesterolemia treatment. Beyond the cholesterol-lowering activity, exploration of neuroprotective properties of this statin indicates its potential benefit in central nervous disorders. The aim of the present study was to assess the effects of atorvastatin in development and expression of morphine-induced analgesic tolerance in male mice and probable involvement of nitric oxide. Chronic and acute treatment with atorvastatin 10 and 20mg/kg, respectively, could alleviate morphine tolerance in development and expression phases. Chronic co-administration of nitric oxide synthase (NOS) inhibitors including L-NAME (non selective NOS inhibitor; 2mg/kg), aminoguanidine (selective inducible NOS inhibitor; 50mg/kg) and 7-NI (selective neuronal NOS inhibitor; 15mg/kg) with atorvastatin blocked the protective effect of atorvastatin in tolerance reversal. Moreover, reversing the atorvastatin effect was also observed in acute simultaneous treatment of L-NAME (5mg/kg) and aminoguanidine (100mg/kg) with atorvastatin. Co-treatment of guanylyl cyclase inhibitor, ODQ (chronic dose: 10mg/kg and acute dose: 20mg/kg) was associated with prevention of atorvastatin anti-tolerance properties. Our results revealed that the atorvastatin modulating role in morphine antinociceptive tolerance is mediated at least in part via nitric oxide in animal pain models of hot plate and tail flick.

Antidepressant effect of pramipexole in mice forced swimming test: A cross talk between dopamine receptor and NMDA/nitric oxide/cGMP pathway.

Pramipexole is a dopamine D2 receptor agonist indicated for treating Parkinson disorder. This study was aimed to investigate the effect of pramipexole in forced swimming test (FST) in mice and the possible involvement of activation of D2 receptors and inhibition of N-methyl-d-aspartate (NMDA) receptors and nitric oxide-cyclic guanosine monophosphate (NO-cGMP) on this effect. Intraperitoneal administration of pramipexole (1-3mg/kg) reduced the immobility time in the FST similar to fluoxetine (20mg/kg, i.p.). This effect of pramipexole (1mg/kg, i.p.) was ceased when mice were pretreated with haloperidol (0.15mg/kg, i.p,) and sulpiride (5mg/kg, i.p) as D2 receptor antagonists, NMDA (75mg/kg,i.p.), l-arginine (750mg/kg, i.p., a substrate for nitric oxide synthase) or sildenafil (5mg/kg, i.p., a phosphodiesterase 5 inhibitor). The administration of MK-801 (0.05mg/kg, i.p., a NMDA receptor antagonist) l-NG-Nitro arginine methyl ester (l-NAME, 10mg/kg, i.p., a non-specific nitric oxide synthase (NOS) inhibitor), 7-nitroindazole (30mg/kg, i.p., a neuronal NOS inhibitor) and methylene blue (10mg/kg, i.p.), an inhibitor of both NOS and soluble guanylyl cyclase (sGC) in combination with the sub-effective dose of pramipexole (0.3mg/kg, i.p.) reduced the immobility. Altogether, our data suggest that the antidepressant-like effect of pramipexole is dependent on the activation of D2 receptor and inhibition of either NMDA receptors and/or NO-cGMP synthesis. These results contribute to the understanding of the mechanisms underlying the antidepressant-like effect of pramipexole and reinforce the role of D2 receptors, NMDA receptors and l-arginine-NO-GMP pathway in the antidepressant mechanism of this agent.

Involvement of Inflammatory Cytokines in Antiarrhythmic Effects of Clofibrate in Ouabain-Induced Arrhythmia in Isolated Rat Atria.

Considering the cardioprotective and anti-inflammatory properties of clofibrate, the aim of the present experiment was to investigate the involvement of local and systemic inflammatory cytokines in possible antiarrhythmic effects of clofibrate in ouabain-induced arrhythmia in rats. Rats were orally treated with clofibrate (300 mg/kg), and ouabain (0.56 mg/kg) was administered to animals intraperitoneally. After induction of anesthesia, the atria were isolated and the onset of arrhythmia and asystole was recorded. The levels of inflammatory cytokines in atria were also measured. Clofibrate significantly postponed the onset of arrhythmia and asystole when compared to control group (P ≤ 0.05 and P ≤ 0.01, resp.). While ouabain significantly increased the atrial beating rate in control group (P ≤ 0.05), same treatment did not show similar effect in clofibrate-treated group (P > 0.05). Injection of ouabain significantly increased the atrial and systemic levels of all studied inflammatory cytokines (P ≤ 0.05). Pretreatment with clofibrate could attenuate the ouabain-induced elevation of IL-6 and TNF-α in atria (P ≤ 0.01 and P ≤ 0.05, resp.), as well as ouabain-induced increase in IL-6 in plasma (P ≤ 0.05). Based on our findings, clofibrate may possess antiarrhythmic properties through mitigating the local and systemic inflammatory factors including IL-6 and TNF-α.

Proconvulsant effect of post-weaning social isolation stress may be associated with dysregulation of opioid system in the male mice.

Opioid system has been reported to be involved in the consequences of post-weaning social isolation stress (SIS) such as hypoalgesia and social behaviors. Also, previous studies have shown that SIS increases mu opioid receptor expression in the regions of the brain associated with epileptogenesis such as basolateral amygdala and cortex. Interestingly, experiencing SIS increases seizure risk in the adulthood. Regarding the SIS-induced alterations in the opioid system, we hypothesize that increase in opioidergic system activity (mostly by mu receptor) may be associated with increase in vulnerability to seizures. In non-stressed mice, morphine at low doses (1 mg/kg) has an anticonvulsant effect on seizure threshold while higher doses (60 mg/kg) are proconvulsant. To support the hypothesis, we showed that administration of anticonvulsant dose of morphine (1 mg/kg) to socially isolated male mice not only was not able to reverse the negative effect of SIS on seizure susceptibility to pentylenetetrazole but also enhanced it. These results support our hypothesis that proconvulsant effect of post-weaning social isolation stress may be associated with dysregulation of opioid system in the adult male mice.

Involvement of the nitrergic system in the proconvulsant effect of social isolation stress in male mice.

Social isolation stress (SIS) in adolescence is accompanied by neurobehavioral disturbances and pathophysiological changes in certain regions of the CNS such as the hippocampus. In this study, we tested whether SIS impacts seizure susceptibility in postnatal male mice due to a role of hippocampal nitric oxide (NO). To do this, we used the pentylenetetrazole (PTZ) model of clonic seizures, open-field test, hole-board test, forced swimming test, and plasma corticosterone assay. We aimed to evaluate if 4 weeks of SIS is capable of decreasing seizure threshold along with altering affective and neuroendocrine responses in isolated conditioned (IC) animals in comparison with socially conditioned (SC) animals. In addition, we applied subeffective doses of NO precursor L-arginine (25, 50, and 100mg/kg) and NOS inhibitors 7-NI (15 and 40 mg/kg), aminoguanidine (50 and 100mg/kg), and L-NAME (10 and 15 mg/kg) to both IC and SC groups prior to the determination of seizure threshold. Injection of a single dose of all mentioned drugs did not induce changes in seizure threshold of SC mice. On the other hand, L-NAME and 7-NI, but not aminoguanidine, modulated the proconvulsant effect of SIS, while L-arginine augmented the latter effect. We also measured the hippocampal nitrite levels after the administration of the aforementioned drugs. Social isolation stress increased the nitrite levels in comparison with those in SC mice, whereas 7-NI and L-NAME, unlike aminoguanidine, mitigated the effect of SIS. Additionally, L-arginine boosted the effects of SIS on nitrite production. In summary, we showed that SIS enhanced seizure susceptibility in the PTZ model of clonic seizures through the activation of the nitrergic system in the hippocampus. Also, we proved that nNOS, but not iNOS, accounts for these changes following SIS.

Anti-pruritic activity of pioglitazone on serotonin-induced scratching in mice: possible involvement of PPAR-gamma receptor and nitric oxide.

Pioglitazone is a member of peroxisome proliferator-activated receptor gamma (PPARγ) agonists, particularly used in management of type II diabetes. However it also has effects in some dermatological disorders. The current study was designed to investigate the effects of oral administration of pioglitazone and the association of nitric oxide, in serotonin-induced scratching in mice. In order to produce the scratching activity, serotonin (141 nm/site) was administered intradermally in the nape of the neck. Pioglitazone in concentrations of 10, 20, 40 and 80 mg/kg, was peroral administered (p.o) as a single dose, 4 h before the serotonin injection. PPAR-γ antagonist, GW9662 (2 mg/kg, i.p); a non-specific nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME; 1 mg/kg, i.p); or a nitric oxide precursor, L-arginine (100 mg/kg, i.p.); adminstrated 15 min before pioglitazone were analyzed for anti-scratching activity. Results obtained showed that pioglitazone (40 and 80 mg/kg, p.o) reduced the scratching in a dose-dependent manner. GW9662 inverted the anti-scratching effect of pioglitazone (80 mg/kg). Acute dose of L-NAME (1 mg/kg, i.p) also prevented the anti-scratching property of pioglitazone (80 mg/kg, p.o); although L-arginine was used in sub-effective dose (100 mg/kg, i.p), however it potentiated the anti-scratching behavior when co-injected with pioglitazone (20 mg/kg, p.o). The results indicate that acute pioglitazone has an anti-scratching effect on serotonin-induced scratching in mice. It is concluded that anti-scratching outcome of acute pioglitazone is initiated via activation of PPAR-γ receptor and to some extent by the NO pathway.

Degradation of phenanthrene and pyrene in spiked soils by single and combined plants cultivation.

The present study was conducted to investigate the capability of four plant species (tall fescue, ryegrass, alfalfa, and rape seed) grown alone and in combination to the degradation of phenanthrene and pyrene (polycyclic aromatic hydrocarbons, PAHs) in spiked soil. After 65 days of plant growth, plant biomass, dehydrogenase activity, water-soluble phenolic (WSP) compounds, plant uptake and accumulation and residual concentrations of phenanthrene and pyrene were determined. Our results showed that presence of vegetation significantly enhanced the dissipation of phenanthrene and pyrene from contaminated soils. Higher degradation rates of PAHs were observed in the combined plant cultivation (98.3-99.2% phenanthrene and 88.1-95.7% pyrene) compared to the single plant cultivation (97.0-98.0% phenanthrene and 79.8-86.0% pyrene). Contribution of direct plant uptake and accumulation of phenanthrene and pyrene was very low compared to the plant enhanced dissipation. By contrast, plant-promoted biodegradation was the predominant contribution to the remediation enhancement. The correlation analysis indicates a negative relation between biological activities (dehydrogenase activity and WSP compounds) and residual concentrations of phenanthrene and pyrene in planted soils. Our results suggest that phytoremediation could be a feasible choice for PAHs contaminated soil. Moreover, the combined plant cultivation has potential to enhance the process.