An Amphiphilic Cell-Penetrating Macrocycle for Efficient Cytosolic Delivery of Proteins, DNA, and CRISPR Cas9.

The discovery of safe platforms that can circumvent the endocytic pathway is of great significance for biological therapeutics that are usually degraded during endocytosis. Here we show that a self-assembled and dynamic macrocycle can passively diffuse through the cell membrane and deliver a broad range of biologics, including proteins, CRISPR Cas9, and ssDNA, directly to the cytosol while retaining their bioactivity. Cell-penetrating macrocycle CPM can be easily prepared from the room temperature condensation of diketopyrrolopyrrole lactams with diamines. We attribute the high cellular permeability of CPM to its amphiphilic nature and chameleonic properties. It adopts conformations that partially bury polar groups and expose hydrophobic side chains, thus self-assembling into micellar-like structures. Its superior fluorescence makes CPM trackable inside cells where it follows the endomembrane system. CPM outperformed commercial reagents for biologics delivery and showed high RNA knockdown efficiency of CRISPR Cas9. We envisage that this macrocycle will be an ideal starting point to design and synthesize biomimetic macrocyclic tags that can readily facilitate the interaction and uptake of biomolecules and overcome endosomal digestion.

Investigating dysregulation of TGF-ß1/SMAD3 signaling in Atopic Dermatitis: A Molecular and Immunohistochemical Analysis".

Atopic Dermatitis (AD) is a persistent and recurring inflammatory condition affecting the skin. An expanding corpus of evidence indicates the potential participation of TGF-ß1 in the modulation of inflammation and tissue remodeling in AD. The primary objective of this study was to examine the aberrant modulation of TGF-ß1/SMAD3 signaling through a comprehensive analysis of their molecular and protein expression profiles. The study encompassed an aggregate of 37 participants, which included 25 AD patients and 12 controls. The assessment of mRNA and protein levels of TGF-ß1 and SMAD3 was conducted utilizing quantitative real-time PCR and immunohistochemistry, whereas serum IgE and vitamin D levels were estimated by ELISA and chemiluminescence, respectively. Quantitative analysis demonstrated a 2.5-fold upregulation of TGF-ß1 mRNA expression in the lesional AD skin (p<0.0001). Immunohistochemistry also exhibited a comparable augmented pattern, characterized by moderate to strong staining intensities. In addition, TGF-ß1 mRNA showed an association with vitamin D deficiency in serum (p<0.02), and its protein expression was linked with the disease severity (p<0.01) Furthermore, a significant decrease in the expression of the SMAD3 gene was observed in the affected skin (p = 0.0004). This finding was further confirmed by evaluating the protein expression and phosphorylation of SMAD3, both of which exhibited a decrease. These findings suggest that there is a dysregulation in the TGF-ß1/SMAD3 signaling pathway in AD. Furthermore, the observed augmentation in mRNA and protein expression of TGF-ß1, along with its correlation with the disease severity, holds considerable clinical significance and emphasizes its potential role in AD pathogenesis.

Comparative study of TNF-α and vitamin D reveals a significant role of TNF-α in NSCLC in an ethnically conserved vitamin D deficient population.

Growing evidence has implicated tumor necrosis factor-alpha (TNF-α) as an important regulator of the tumor microenvironment. Moreover, various molecular epidemiological studies have proposed vitamin D deficiency to be a mediator of cancer progression. Here we comparatively analyzed the role of TNF-α and vitamin D in non-small cell lung cancer (NSCLC) in an ethnically conserved vitamin D deficient population. Confirmed NSCLC cases (n = 190) matchedfor age, gender, dwelling, and smoking against cancer-free healthy controls ((n = 200) were genotyped for TNF-α promoter polymorphisms (rs361525 and rs1800629) by PCR-RFLP. 48 NSCLC tumor and adjacent normal tissues were quantified for TNF-α mRNA expression by RT-qPCR. 48 NSCLC cases and 60 healthy controls were analyzed for TNF-α and vitamin D serum levels by ELISA and chemiluminescence respectively. Our study indicates thatrs361525 and rs1800629 bear a significant risk towards NSCLC. Both mutant genotype and mutant allele of rs361525 elicit a likelihood of NSCLC reflected by their odds ratio (OR) of 3.16 and 1.81 respectively. In case of rs1800629, the heterogeneous genotype (GA) showed two fold higher risk for NSCLC (OR-2.07, P = 0.006), which could be attributed to the presence of the mutant allele as reflected by overall frequency of mutant A allele vs wild G allele (OR-1.92, P = 0.01). A combined effect of genotypes for rs361525 and rs1800629 revealed a 3.7 fold higher risk towards NSCLC for the presence of heterozygous genotype at both loci. Our preliminary expression results showed significant increase of TNF-α mRNA expression in tumor tissues of NSCLC as compared to adjacent normal tissues [cases- 8.56 ± 3.90vs controls-4.88 ± 2.96, P < 0.0001)] which was further affirmed by extrapolation of TNF-α expression in serum (Cases- 55.75 ± 22.50vs controls- 21.46 ± 27.75, P < 0.0001). Multivariate regression analyses revealed TNF-α mRNA expression to be significantly associated with NSCLC cases less than 50 years of age (P < 0.05). In comparison to the putative role of TNF-α in NSCLC as suggested by the results observed, vitamin D showed no significance towards any of the analyzed parameters or with the risk of NSCLC. This study suggests that TNF-α could be a potential mediator of NSCLC which bears important clinical implications and could be an important therapeutic marker in NSCLC.

Supramolecular Interaction of Molecular Cage and ß-Galactosidase: Application in Enzymatic Inhibition, Drug Delivery and Antimicrobial Activity.

Enzyme inhibitors play a crucial role in diagnosis of a wide spectrum of diseases related to bacterial infections. We report here the effect of a water-soluble self-assembled PdII8 molecular cage towards ß-galactosidase enzyme activity. The molecular cage is composed of a tetrapyridyl donor (L) and cis-[(en)Pd(NO3 )2 ] (en=ethane-1,2-diamine) acceptor and it has a hydrophobic internal cavity. We have observed that the acceptor moiety mainly possesses the ability to inactivate the ß-galactosidase enzyme activity. Kinetic investigation revealed the mixed mode of inhibition. This inhibition strategy was extended to control the growth of methicillin-resistant Staphylococcus aureus. The internalization of the Pd(II) cage inside the bacteria was confirmed when bacterial solutions were incubated with curcumin loaded cage. The intrinsic green fluorescence of curcumin made the bacteria glow when put under an optical microscope. Furthermore, this curcumin loaded molecular cage shows an enhanced antibacterial activity. Thus, PdII8 molecular cage is quite attractive due to its dual role as enzyme inhibitor and drug carrier.

Coordination-Driven Self-Assembly of Discrete Molecular Nanotubular Architectures.

Two new M8L4 tetrafacial nanotubes (T1 and T3) of different lengths have been synthesized in water using ligands L1 and L2, respectively, with acceptor cis-[(dch)Pt(NO3)2] (M) using coordination-driven self-assembly [where dch is 1,2-diaminocyclohexane, L1 is 1,4-di(pyrimidin-5-yl)benzene, and L2 is 4,4'- di(pyrimidin-5-yl)-1,1'-biphenyl]. In addition to complex T1, a tetrahedral cage of composition [M12(L1)6] (T2) was also formed in the self-assembly reaction of ligand L1 with cis-[(dch)Pt(NO3)2]. The precise composition of the products (T1 and T2) in solution was confirmed by 1H NMR and ESI-MS. Pure tube T1 was separated out by a crystallization technique and fully characterized by 1H NMR and X-ray diffraction. Temperature- and concentration-dependent NMR studies indicated no equilibrium between T1 and T2 in the solution phase, and the proportion of T1 and T2 in the mixture depends on the temperature of the reaction. In contrast to ligand L1, the self-assembly of the longer ligand, L2, with cis-[(dch)Pt(NO3)2] gave only tetrafacial tube [M8(L2)4] (T3) without any tetrahedral cage.

A Discrete Self-Assembled Pd12 Triangular Orthobicupola Cage and its Use for Intramolecular Cycloaddition.

Water-soluble Pd12 L6 coordination cage TC-1 was synthesized by coordination-driven self-assembly of symmetrical tetrapyridyl donor L with 90° ditopic acceptor cis-[Pd(NO3 )2 (tmeda)] [tmeda=N,N,N',N'-tetramethylethane-1,2-diamine]. The Pd12 L6 coordination assembly is an uncommon example of a coordination cage having triangular orthobicupola-like geometry. It was characterized by multinuclear NMR spectroscopy, ESI-MS, and single-crystal X-ray diffraction. Self-assembly of a tetratopic donor with a cis-blocked 90° ditopic acceptor generally yields tri-/tetra-/hexagonal barrels or closed cubic cages. However, in the present case the donor and acceptor are arranged in an unusual fashion to generate an orthobicupola geometry in which two triangular cupola share a common irregular hexagonal face. The cage was used to perform intramolecular cycloaddition reactions of O-propargylated benzylidinebarbituric acid derivatives in nitromethane. Several penta-/tetracyclouracil derivatives were synthesized through cage-catalyzed [4+2] cycloaddition reactions in a concerted manner with good to high conversion under mild reaction conditions, whereas in the absence of cage TC-1 similar reactions led to lower conversion to the cyclized products in organic solvent. This approach is of particular importance compared to the literature reports on the synthesis of similar compounds under high-temperature reflux conditions with high catalyst loading.

Water-Soluble Pd8L4 Self-assembled Molecular Barrel as an Aqueous Carrier for Hydrophobic Curcumin.

A tetrafacial water-soluble molecular barrel (1) was synthesized by coordination driven self-assembly of a symmetrical tetrapyridyl donor (L) with a cis-blocked 90° acceptor [cis-(en)Pd(NO3)2] (en = ethane-1,2-diamine). The open barrel structure of (1) was confirmed by single crystal X-ray diffraction. The presence of a hydrophobic cavity with large windows makes it an ideal candidate for encapsulation and carrying hydrophobic drug like curcumin in an aqueous medium. The barrel (1) encapsulates curcumin inside its molecular cavity and protects highly photosensitive curcumin from photodegradation. The photostability of encapsulated curcumin is due to the absorption of a high proportion of the incident photons by the aromatic walls of 1 with a high absorption cross-sectional area, which helps the walls to shield the guest even against sunlight/UV radiations. As compared to free curcumin in water, we noticed a significant increase in solubility as well as cellular uptake of curcumin upon encapsulation inside the water-soluble molecular barrel (1) in aqueous medium. Fluorescence imaging confirmed that curcumin was delivered into HeLa cancer cells by the aqueous barrel (1) with the retention of its potential anticancer activity. While free curcumin is inactive toward cancer cells in aqueous medium at room temperature due to negligible solubility, the determined IC50 value of ∼14 µM for curcumin in aqueous medium in the presence of the barrel (1) reflects the efficiency of the barrel as a potential curcumin carrier in aqueous medium without any other additives. Thus, two major challenges of increasing the bioavailability and stability of curcumin in aqueous medium even in the presence of UV light have been addressed by using a new supramolecular water-soluble barrel (1) as a drug carrier.

Self-Assembly of Discrete RuII8 Molecular Cages and Their in Vitro Anticancer Activity.

Four new octanuclear Ru(II) cages (OC-1-OC-4) were synthesized from dinuclear p-cymene ruthenium(II) acceptors [Ru2(µ-η4-C2O4)(CH3OH)2(η6-p-cymene)2](O3SCF3)2 (A1), [Ru2(µ-η4-C6H2O4)(CH3OH)2(η6-p-cymene)2](O3SCF3)2 (A2), [Ru2(dhnq)(H2O)2(η6-p-cymene)2](O3SCF3)2 (A3), and [Ru2(dhtq)(H2O)2(η6-p-cymene)2](O3SCF3)2 (A4) separately with a tetradentate pyridyl ligand (L1) in methanol using coordination-driven self-assembly [L1= N,N,N',N'-tetra(pyridin-4-yl)benzene-1,4-diamine]. The octanuclear cages are fully characterized by various spectroscopic techniques including single-crystal X-ray diffraction analysis of OC-4. The self-assembled cages show strong in vitro anticancer activity against human lung adenocarcinoma A549 and human cervical cancer HeLa cell lines as observed from the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Of all the octanuclear cages, OC-3 exhibits remarkable anticancer activity against both cancer cell lines and is more active than that reported for cisplatin. The excellent anticancer activity of OC-3 and OC-4 highlights the importance of the synergistic effects of the spacer component of the dinuclear p-cymene Ru(II) acceptor clips.

Association of IL1 beta gene polymorphism and allograft functions in renal transplant recipients :a case control study from Kashmir Valley.

BACKGROUND: Cytokines have been found to be the important mediators during renal graft outcome. Therefore, we designed this study to investigate the role of recipients' IL-1 ß promoter (-511) and IL-1 ß exon-5 (+3954) polymorphisms with the risk of graft outcome. METHODOLOGY: We enrolled one hundred recipients of living-related renal transplants together with the age and sex matched controls from the healthy population not having any renal abnormality for this study. Genotype frequencies of the IL-1 ß promoter (-511) and IL-1 ß exon-5 (+3954) were analyzed using PCR-RFLP technique. RESULTS: Our results revealed significant differences in the healthy control group and patient group in IL 1ß +3954 (p < 0.001). The frequency of variant type TT genotype was higher in RE group as compared to SGF and showed 4 fold risk of rejection (OR = 4.54, p < 0.069) although p value was not significant. The frequency of wild type CC genotype and CT was not significant (p value 0.89 and 0.74 respectively). CONCLUSION: Our findings suggest that there is a prevalence of mutated allele of IL-1 gene cluster in our population, which may be responsible for renal dysfunction.

Synthesis, molecular docking and evaluation of antifungal activity of Ni(II), Co(II) and Cu(II) complexes of porphyrin core macromolecular ligand.

Porphyrin core dendrimeric ligand (L) was synthesized by Rothemund synthetic route in which p-hydroxy benzaldehyde and pyrrole were fused together. The prepared ligand was complexed with Ni(II), Cu(II) and Co(II) ions, separately. Both the ligand and its complexes were characterized by elemental analysis and spectroscopic studies (FT-IR, UV-Vis, (1)HNMR). Square planar geometries were proposed for Cu(II), Ni(II) and Co(II) ions in cobalt, Nickel and copper complexes, respectively on the basis of UV-Vis spectroscopic data. The ligand and its complex were screened on Candida albicans (ATCC 10231), Aspergillus fumigatus (ATCC 1022), Trichophyton mentagrophytes (ATCC 9533) and Pencillium marneffei by determining MICs and inhibition zones. The activity of the ligand and its complexes was found to be in the order: CuL ˃ CoL ≈ NiL ˃ L. Detection of DNA damage at the level of the individual eukaryotic cell was observed by commet assay. Molecular docking technique was used to understand the ligand-DNA interactions. From docking experiment, we conclude that copper complex interacts more strongly than rest two.

A Pd24 Pregnant Molecular Nanoball: Self-Templated Stellation by Precise Mapping of Coordination Sites.

We found that Pd(II) ion (M) and the smallest 120° bidentate donor pyrimidine (L(a)) self-assemble into a mononuclear M(L(a))4 complex (1a) instead of the expected smallest M12(L(a))24 molecular ball (1), presumably due to the weak coordination nature of the pyrimidine. To construct such a pyrimidine bridged nanoball, we employed a new donor tris(4-(pyrimidin-5-yl)phenyl)amine (L); which upon selective complexation with Pd(II) ions resulted in the formation of a pregnant M24L24 molecular nanoball (2) consisting of a pyrimidine-bridged Pd12 baby-ball supported by a Pd12 larger mother-ball. The formation of the baby-ball was not successful without the support of the mother-ball. Thus, we created an example of a self-assembly where the inner baby-ball resembling to the predicted M12(L(a))24 ball (1) was incarcerated by the giant outer mother-ball by means of geometrical constraints. Facile conversion of the pregnant ball 2 to a smaller M12(L(b))24 ball 3 with dipyridyl donor was achieved in a single step.

Significant impact of IL-6 -174G/C but inverse relation with -634 C/G polymorphism in patients with non-small cell lung cancer in Kashmiri population.

To study the possible role of proinflammatory interleukin 6 -174 G>C (rs 1800795) and -634 C>G (rs 1800796) polymorphism in the pathogenesis of non-small cell lung cancer (NSCLC). A total of 190 NSCLC patients and 200 healthy controls were evaluated for polymorphic analysis of -174 G/C and -634 C/G by PCR-RFLP followed by DNA sequencing. A significant association was observed in the genotypic and allelic distribution of IL-6 -174 G/C in the NSCLC group as compared to control group [OR = 2.7 (1.77-4.11), p < 0.0001]. Smokers with the -174C allele were found to be significantly associated with NSCLC (p = 0.01), while 634C/G SNP showed an inverse relation [OR-0.4, p < 0.0001]. The present investigation revealed a significant association of the IL6 -174 G/C gene promoter polymorphism with NSCLC, and thus, the IL-6 -174G/C genotype can be considered as one of the biological markers in the etiology of NSCLC.

COX-2 overexpression and -8473 T/C polymorphism in 3' UTR in non-small cell lung cancer.

A new class of compounds targeting cyclooxygenase 2 (COX-2) together with other different clinically used therapeutic strategies has recently shown a promise for the chemoprevention of several solid tumors including lung cancer. The aim was to study the possible role of COX-2 -8473 T/C NP and its expression in the pathogenesis of non-small cell lung cancer. One hundred ninety non-small cell lung cancer (NSCLC) patients and 200 healthy age-, sex-, and smoking-matched controls were used for polymorphic analysis, and 48 histopathologically confirmed NSCLC patients were analyzed for COX-2 messenger RNA (mRNA) and protein expression. Our results showed that the frequencies of variant genotypes 8473 CT/CC were significantly less common in the cases (30.0%) than in the controls (36%), suggesting that the 8473 C variant allele is related with lower susceptibility in NSCLC (OR = 0.79, 95% CI 0.54-1.4). However, the frequency of COX-2 -8473 TC and CC genotypes were significantly associated with age in NSCLC (P = 0.02). Quantitative real-time expression analysis showed a significant increase in the COX-2 mRNA in tumor tissues as compared to their adjacent normal tissues [delta cycle threshold (ΔCT) = 9.25 ± 4.67 vs 5.63 ± 3.85, P = 0.0001]. Multivariate logistic regression analyses revealed that the COX-2 expression was associated significantly with age (P = 0.044). Also, an increasing trend was observed in stages I and II and in female patients compared to stages III and IV and male patients, respectively, but no statistical significance was observed. However, COX-2 mRNA expression shown no association with the -8473 C variant allele. Our findings indicate that the COX-2 T8473C polymorphism may contribute to NSCLC cancer susceptibility in the Kashmiri population, while our expression analysis revealed a significant increase of COX-2 in tumor tissues as compared to their adjacent normal tissues, suggesting that it could become an important therapeutic marker in NSCLC in the future.

Stroke-induced immune depression-a randomized case control study in Kashmiri population of North India.

BACKGROUND: Stroke leads to transient immunedepression, which leads to increased incidence of poststroke infections. Because infection is one of the most common causes of increased mortality in patients with stroke, this study was undertaken to document immunedepression after stroke in our population. METHODS: A case-controlled study wherein 39 patients with acute ischemic stroke in the age group of 18 and 60 years without any evidence of previous immunedepression were included. Interleukin 6 (IL-6) and interleukin 10 (IL-10) levels were checked in plasma in both the groups on day 3 and day 45. Also Cortisol and epinephrine levels were checked in the urine samples collected on day 3 and day 8. RESULTS: No significant difference was seen between the IL-6 and the IL-10 levels in samples collected on day 3 between the controls and cases, whereas Cortisol and norepinephrine were significantly raised in samples collected on day 3 in cases who developed infection as compared with controls. CONCLUSIONS: The higher levels of urinary cortisol and norepinephrine were observed in patients with stroke who developed infections, which indirectly reflected increased amount of stroke related stress. Furthermore, the levels of plasma IL-6 and IL-10 were also elevated in the same group of patients, which means transformation of immunecompetence to immunedepression, which is responsible for higher mortality. Subsequently on recovery from infection the plasma levels of interleukins and urinary cortisol and norepinephrine did not show any difference, which indirectly means recovery of the immune system on recovery from acute stage of stroke. Mortality in the patients with infection was increased than controls.

Anti-lung Cancer Activity of Synthesized Substituted 1,4-Benzothiazines: An Insight from Molecular Docking and Experimental Studies.

BACKGROUND: Thiazine, a 6-membered distinctive heterocyclic motif with sulfur and nitrogen atoms, is one of the heterocyclic compounds that functions as a core scaffold in a number of medicinally significant molecules. Small thiazine-based compounds may operate simultaneously on numerous therapeutic targets and by employing a variety of methods to halt the development, proliferation, and vasculature of cancer cells. We have, herein, reported a series of substituted 1,4 benzothiazines as potential anticancer agents for the treatment of lung cancer. METHODS: In order to synthesize 2,3-disubstituted-1,4 benzothiazines in good yield, a facile green approach for the oxidative cycloaddition of 2-amino benzenethiol and 1,3-dicarbonyls employing a catalytic amount of ceric ammonium nitrate has been devised. All the molecules have been characterized by spectral analysis and tested for anticancer activity against the A-549 lung cancer cell line using various functional assays. Further in silico screening of compound 3c against six crucial inflammatory molecular targets, such as Il1-α (PDB ID: 5UC6), Il1- ß (PDB ID: 6Y8I), Il6 (PDB ID: 1P9M), vimentin (PDB ID: 3TRT), COX-2 (PDB ID: 5KIR), Il8 (PDB ID: 5D14), and TNF-α (PDB ID: 2AZ5), was done using AutoDock tool. RESULTS: Among the synthesized compounds, propyl 3-methyl-3,4-dihydro-2H-benzo[b][1,4]thiazine-2- carboxylate (3c) was found to be most active based on cell viability assays using A-549 lung cancer cell line and was found to effectively downregulate various pro-inflammatory genes, like Il1-α, Il1-ß, Il6, vimentin, COX-2, Il8, and TNF-α in vitro. The ability of the molecule to effectively suppress the proliferation and migration of lung cancer cells in vitro has been further demonstrated by the colony formation unit assay and wound healing assay. Molecular docking analysis showed the maximal binding affinity (- 7.54 kcal/mol) to be exhibited by compound 3c against IL8. CONCLUSION: A green unconventional route for the synthesis of 2,3-disubstituted-1,4 benzothiazines has been developed. All the molecules were screened for their activity against lung cancer and the data suggested that the presence of an additional unbranched alkyl group attached to the thiazine ring increased their activity. Also, in vitro and in silico modeling confirmed the anti-cancer efficiency of compound 3c, encouraging the exploration of such small molecules against cancer.

Role of vitamin D receptor (VDR) polymorphisms in susceptibility to multiple myeloma in ethnic Kashmiri population.

BACKGROUND: Vitamin D regulates many biological processes including bone metabolism, innate immune response, and cell proliferation and differentiation by binding to its receptor VDR. Vitamin D receptor (VDR) gene polymorphisms have been associated with many cancers like breast, colorectal, prostate, and skin. The main aim of this study was to determine whether VDR polymorphisms (ApaI, BsmI and FokI) are associated with increased risk of multiple myeloma. METHODS: We designed a case control study where 75 multiple myeloma cases were studied for VDR polymorphisms (ApaI, BsmI and FokI) against 150 controls taken from general population. The polymorphisms of VDR gene were investigated using PCR-RFLP method. RESULTS: We did not find any significant association between ApaI and BsmI polymorphisms and multiple myeloma risk (P>0.05), but FokI polymorphism was significantly associated with increased risk for multiple myeloma. We also found a significant association between the ff variant genotype with creatinine levels, albumin levels, and Durie-Salmon stage III. CONCLUSION: Our findings suggest that the FokI polymorphism is involved in the increased susceptibility to development and progression in multiple myeloma in the ethnic Kashmiri population. Furthermore these results suggest that ff genotype is associated with higher risk for developing multiple myeloma.

Effect of in situ mixed micellization of ester-functionalized gemini surfactant at different pHs on solubilization and cosolubilization of various polycyclic aromatic hydrocarbons of varying hydrophobicities.

A pH controlled cleavability unfolds the 3-in-1 surfactant feature of an ester-bonded gemini surfactant, 2, 2'-[(oxybis (ethane-1,2-diyl))bis (oxy)]bis (N-hexadecyl-N,Ndimethyl-2-oxoethanaminium) dichloride (C16-C4O2-C16), by reinforcing in-situ mixed micellization between cleaved components at non-neutral pH (pH 3,12). The triplicity is assigned to two mixed-micelle variants at pH 3 and pH 12 besides the unhydrolyzed C16-C4O2-C16 at pH 7. The pH-controlled aggregation of such trichotomic surfactant dramatically enhances the micellar solubilization/cosolubilization of PAHs viz. naphthalene (Np), phenanthrene (Ph), pyrene (Py), perylene (Pe). The cosolubilization of binary/ternary PAH mixtures in such remarkable micellar assemblies at pH 3, 7 and 12 yields intriguing synergistic or antagonistic solubility outcomes correlated to PAH-PAH and PAH-micelle interactions. This study provides valuable insights into the potential applications of the ester-bonded gemini surfactant for the cosolubilization of undesirable hydrophobic compounds at natural sites having variable pH.

Analysis of intronic SNP (rs4147358) and expression of SMAD3 gene in Atopic Dermatitis: A case-control study.

BACKGROUND: Atopic Dermatitis (AD) is a multifactorial cutaneous disorder associated with chronic inflammation of the skin. Growing evidence points to TGF-ß/SMAD signaling as a key player in mediating inflammation and the subsequent tissue remodeling, often resulting in fibrosis. This study investigates the role of a core transcription factor involved in TGF-ß signaling i.e., SMAD3 genetic variants (rs4147358) in AD predisposition and its association with SMAD3 mRNA expression, serum IgE levels, and sensitization to various allergens in AD patients. METHODS: A total of 246 subjects including 134 AD cases and 112 matched healthy controls were genotyped for SMAD3 intronic SNP by PCR-RFLP. mRNA expression of SMAD3 was determined by quantitative Real-Time PCR (qRT-PCR), Vitamin-D levels by chemiluminescence, and total serum IgE levels by ELISA. In-vivo allergy testing was performed for the evaluation of allergic reactions to house dust mites (HDM) and food allergens. RESULTS: A significantly higher frequency of mutant genotype AA (cases: 19.4% vs controls: 8.9%) (OR = 2.8, CI = 1.2 - 6.7, p = 0.01) was observed in AD cases. The mutant allele 'A' also showed a 1.9-fold higher risk for AD compared to the wild allele 'C' indicating that the carriers of the A allele have a higher risk for AD predisposition (OR-1.9, CI = 1.3-2.8, p < 0.001). In addition, quantitative analysis of SMAD3 mRNA in peripheral blood showed 2.8-fold increased expression in AD cases as compared to healthy controls. Stratification analysis revealed the association of the mutant AA genotype with deficient serum Vitamin D levels (p = 0.02) and SMAD3 mRNA overexpression with HDM sensitization (p = 0.03). Furthermore, no significant association of genotypes with SMAD3 mRNA expression was observed. CONCLUSION: Our study indicates that SMAD3 intronic SNP bears a significant risk of AD development. Moreover, overexpression of SMAD3 mRNA and its association with HDM sensitization highlights the possible role of this gene in AD pathogenesis.

Gene variants and mRNA expression analysis of SOCS3 and its association with serum IL-4 levels in atopic diseases.

BACKGROUND: The suppressors of cytokine signaling (SOCS) are a class of negative regulators for several aspects of cytokine signaling that have been attributed to the pathophysiology of inflammatory disorders. Given the role of the SOCS3 gene in regulating Th2 cell proliferation, our study aimed to analyze two SOCS3 SNPs viz. rs8074003 and rs7222391, and their potential influence on IL-4 levels and SOCS3 mRNA expression besides analyzing the interaction of the SOCS3 genotypes with the various clinicopathological parameters. METHODS: A total of 314 subjects including 154 atopic cases and 160 healthy controls were genotyped for SOCS3 polymorphisms by PCR-RFLP. SOCS3 mRNA was quantified by Real-Time PCR. The serum IL-4 and total IgE levels were determined by ELISA and Vitamin-D levels were quantified by chemiluminescence. RESULTS: The CC genotype of rs8074003 was more frequent in atopic cases and posed a 3- fold risk of atopy (p = 0.001) whereas CG and GG genotypes were widespread in the controls (p = 0.1). For the other SNP rs7222391, there was no difference in genotypic and allelic distribution. The SOCS3 mRNA expression and serum IL-4 levels were substantially increased in the atopic cases with a significant positive correlation between them (p < 0.05). CONCLUSION: SOCS3 SNP rs8074003 poses a convincing risk of atopic disease development. The SOCS3 expression and IL-4 levels were up-regulated in total atopy and in its different presentations. It seems plausible to target SOCS3 and IL-4 as a potential target for the diagnosis of atopy and for the development of reliable personalized therapeutic strategies to control atopic conditions.

Metabolic Abnormalities, Inflammatory Markers and Endothelial Dysfunction in Hyperprolactinemia due to Prolactinoma before and after Normalization of Serum Prolactin: A Prospective Case Control Study.

Background: Hyperprolactinemia is associated with obesity, dyslipidemia, insulin resistance, and low-grade inflammation which may promote endothelial dysfunction (EnD). Limited work has been done on EnD in prolactinomas and we, therefore, studied serum markers of inflammation and EnD in patients with prolactinomas before and after treatment with dopamine agonists. Methodology: Fifty-six treatment naïve patients with prolactinomas and fifty-three (apparently healthy age and sex-matched) controls were enrolled in the study and subjected to clinical assessment and laboratory investigations including blood glucose, total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, urea, creatinine, uric acid, erythrocyte sedimentation rate (ESR), highly sensitive C-reactive protein (hsCRP) and markers of EnD i.e., intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Patients were treated with a dopamine agonist (cabergoline) and parameters (like ESR, hsCRP, ICAM-1, and VCAM-1) were measured at 12 weeks. Results: The majority of the patients (84%) were female, more than half (52%) had metabolic syndrome and over a third (36%) were obese. Blood glucose fasting, HbA1c, lipid fractions, ESR, hsCRP, ICAM-1, and VCAM-1 were significantly higher in patients than in controls. Median ICAM-1 was 1331.95 ng/ml (IQR 803.43-1825.99) in patients vs 753.04 ng/ml (IQR 402.04-871.55) in controls, P < 0.001 and median VCAM-1in patients was 971.35 ng/ml (IQR 695.03-1285.23) as against 634.56 ng/ml (IQR 177.49-946.50) in controls, p0.001. Serum ICAM-1 and VCAM-1 correlated positively with hsCRP. On multivariate regression analysis, serum hsCRP was the only significant predictor of change in ICAM-1 and VCAM-1. Normalization of serum PRL with CAB resulted in a significant decrease in metabolic parameters, ESR, hsCRP, ICAM-1, and VCAM-1. Conclusion: Hyperprolactinemia because of prolactinoma is associated with EnD secondary to systemic inflammation and metabolic abnormalities which improve after treatment with DA.