De novo variants in KLF7 are a potential novel cause of developmental delay/intellectual disability, neuromuscular and psychiatric symptoms.

Due to small numbers of reported patients with pathogenic variants in single genes, the phenotypic spectrum associated with genes causing neurodevelopmental disorders such as intellectual disability (ID) and autism spectrum disorder is expanding. Among these genes is KLF7 (Krüppel-like factor 7), which is located at 2q33.3 and has been implicated in several developmental processes. KLF7 has been proposed to be a candidate gene for the phenotype of autism features seen in patients with a 2q33.3q34 deletion. Herein, we report 4 unrelated individuals with de novo KLF7 missense variants who share similar clinical features of developmental delay/ID, hypotonia, feeding/swallowing issues, psychiatric features and neuromuscular symptoms, and add to the knowledge about the phenotypic spectrum associated with KLF7 haploinsufficiency.

Crystallisation-enhanced bulk hole mobility in phenothiazine-based organic semiconductors.

A series of three novel donor-acceptor systems based on C(3)-malononitrile-substituted phenothiazines was synthesised in good overall yields and their thermal, spectroscopic, and electrochemical properties were characterised. The compounds were prepared through a sequence of Ullmann-coupling, Vilsmeier-Haack formylation and Knoevenagel-condensation, followed by Suzuki-coupling reactions for introduction of aryl substitutents at C(7) position of the phenothiazine. The introduction of a donor unit at the C(7) position exhibited a weak impact on the optical and electrochemical characteristics of the compounds and led to amorphous films with bulk hole mobilities in the typical range reported for phenothiazines, despite the higher charge delocalisation as attested by computational studies. In contrast, highly ordered films were formed when using the C(7)-unsubstituted 3-malononitrile phenothiazine, exhibiting an outstanding mobility of 1 × 10-3 cm2 V-1 s-1, the highest reported for this class of compounds. Computational conformational analysis of the new phenothizanes suggested that free rotation of the substitutents at the C(7) position suppresses the ordering of the system, thereby hampering suitable packing of the new materials needed for high charge carrier mobility.

Further evidence that de novo missense and truncating variants in ZBTB18 cause intellectual disability with variable features.

Identification of rare genetic variants in patients with intellectual disability (ID) has been greatly accelerated by advances in next generation sequencing technologies. However, due to small numbers of patients, the complete phenotypic spectrum associated with pathogenic variants in single genes is still emerging. Among these genes is ZBTB18 (ZNF238), which is deleted in patients with 1q43q44 microdeletions who typically present with ID, microcephaly, corpus callosum (CC) abnormalities, and seizures. Here we provide additional evidence for haploinsufficiency or dysfunction of the ZBTB18 gene as the cause of ID in five unrelated patients with variable syndromic features who underwent whole exome sequencing revealing separate de novo pathogenic or likely pathogenic variants in ZBTB18 (two missense alterations and three truncating alterations). The neuroimaging findings in our cohort (CC hypoplasia seen in 4/4 of our patients who underwent MRI) lend further support for ZBTB18 as a critical gene for CC abnormalities. A similar phenotype of microcephaly, CC agenesis, and cerebellar vermis hypoplasia has been reported in mice with central nervous system-specific knockout of Zbtb18. Our five patients, in addition to the previously described cases of de novo ZBTB18 variants, add to knowledge about the phenotypic spectrum associated with ZBTB18 haploinsufficiency/dysfunction.

Design of selective TACE inhibitors using molecular docking studies: Synthesis and preliminary evaluation of anti-inflammatory and TACE inhibitory activity.

Tumor necrosis factor-α (TNF-α) converting enzyme (TACE) has been considered one of the principal therapeutic targets for the treatment of TNF-dependent pathologies. Several TACE inhibitors have been reported, but none of them has been successfully passed to phase II clinical trials. In the present work, we attempted to design highly selective new non-hydroxamate sulfonamide TACE inhibitors. The docking study was performed on one of the crystal structures of TACE, selected based on its resolution and R value, to tackle the flexibility issue of the active site. The results allowed us to distinguish the analogues with a higher binding affinity toward the active site of TACE and to identify the substituent of analogues needed for binding with the surrounding site of the enzyme. Finally the analogues were docked on crystal structures of six different matrix metalloproteinases (MMPs) for a selectivity study of TACE over MMPs. Some of these analogues were synthesized and subjected to preliminary testing for in vivo anti-inflammatory activity and TACE inhibitory activity.

Extended x-ray absorption fine structure measurements on asymmetric bipolar pulse direct current magnetron sputtered Ta(2)O(5) thin films.

Tantalum pentoxide (Ta2O5) thin films have been deposited on fused silica substrates using a novel asymmetric bipolar DC magnetron sputtering technique under a mixed ambient of oxygen and argon. Films have been prepared at different oxygen-to-argon ratios, and the sputtering ambient and optical properties of the films have been investigated by spectroscopic ellipsometry, while the structural analysis of the films has been carried out by grazing incidence x-ray diffraction and extended x-ray absorption fine structure (EXAFS) measurements. The concentration of oxygen and tantalum in the Ta2O5 films has been estimated by Rutherford backscattering spectrometry (RBS). The variation of the optical constants of the films with changes in deposition parameters has been explained in the light of the change in average Ta-O bond lengths and oxygen coordination around Ta sites as obtained from EXAFS measurements. The trend in variation of the oxygen-to-tantalum ratio in the films obtained from RBS measurement, as a function of oxygen partial pressure used during sputtering, is found to follow the trend in variation of the oxygen coordination number around Ta sites obtained from EXAFS measurement.

HAADF-STEM atom counting in atom probe tomography specimens: Towards quantitative correlative microscopy.

The geometry of atom probe tomography tips strongly differs from standard scanning transmission electron microscopy foils. Whereas the later are rather flat and thin (<20 nm), tips display a curved surface and a significantly larger thickness. As far as a correlative approach aims at analysing the same specimen by both techniques, it is mandatory to explore the limits and advantages imposed by the particular geometry of atom probe tomography specimens. Based on simulations (electron probe propagation and image simulations), the possibility to apply quantitative high angle annular dark field scanning transmission electron microscopy to of atom probe tomography specimens has been tested. The influence of electron probe convergence and the benefice of deconvolution of electron probe point spread function electron have been established. Atom counting in atom probe tomography specimens is for the first time reported in this present work. It is demonstrated that, based on single projections of high angle annular dark field imaging, significant quantitative information can be used as additional input for refining the data obtained by correlative analysis of the specimen in APT, therefore opening new perspectives in the field of atomic scale tomography.

Shaking-induced crystallization of dense sphere packings.

We use a hybrid Monte Carlo algorithm to simulate the shaking of spheres at different vibrational amplitudes and find that spontaneous crystallization occurs in specific dynamical regimes. Several crystallizing transitions are typically observed, leading to end states which can be fully or partially ordered, depending on the shaking amplitude, which we investigate using metrics of global and local order. At the lowest amplitudes, crystallization is incomplete, at least for our times of observation. For amplitude ranges where crystallization is complete, there is typically a competition between hcp and fcc ordering. It is seen that fcc ordering typically predominates; in fact for an optimal range of amplitudes, spontaneous crystallization into a pure fcc state is observed. An interesting feature is the breakdown of global order when there is juxtaposition of fully developed hcp and fcc order locally: we suggest that this is due to the interfaces between the different domains of order, which play the same role as dislocations.

Optimization of sample preparation variables for wedelolactone from Eclipta alba using Box-Behnken experimental design followed by HPLC identification.

INTRODUCTION: Coumestan wedelolactone is an important phytocomponent from Eclipta alba (L.) Hassk. It possesses diverse pharmacological activities, which have prompted the development of various extraction techniques and strategies for its better utilization. The aim of the present study is to develop and optimize supercritical carbon dioxide assisted sample preparation and HPLC identification of wedelolactone from E. alba (L.) Hassk. METHODS: The response surface methodology was employed to study the optimization of sample preparation using supercritical carbon dioxide for wedelolactone from E. alba (L.) Hassk. The optimized sample preparation involves the investigation of quantitative effects of sample preparation parameters viz. operating pressure, temperature, modifier concentration and time on yield of wedelolactone using Box-Behnken design. The wedelolactone content was determined using validated HPLC methodology. The experimental data were fitted to second-order polynomial equation using multiple regression analysis and analyzed using the appropriate statistical method. RESULTS: By solving the regression equation and analyzing 3D plots, the optimum extraction conditions were found to be: extraction pressure, 25 MPa; temperature, 56 °C; modifier concentration, 9.44% and extraction time, 60 min. Optimum extraction conditions demonstrated wedelolactone yield of 15.37 ± 0.63 mg/100 g E. alba (L.) Hassk, which was in good agreement with the predicted values. DISCUSSION AND CONCLUSION: Temperature and modifier concentration showed significant effect on the wedelolactone yield. The supercritical carbon dioxide extraction showed higher selectivity than the conventional Soxhlet assisted extraction method.

Analgesic and anti-inflammatory activities of 18-acetoxy-ent-kaur-16-ene from Annona squamosa L. bark.

The Annona squamosa L. bark was collected from Ahmednagar district, India. 18-Acetoxy-ent-kaur-16-ene was isolated from petroleum ether extract (PE) and studied for its analgesic and anti-inflammatory activities. 18-Acetoxy-ent-kaur-16-ene at the doses of 12.5 and 25 mg/kg, and PE at a dose of 50 mg/kg exhibited significant analgesic along with anti-inflammatory activity.

Stability indicating LC method for simultaneous determination of irbesartan and hydrochlorothiazide in pharmaceutical preparations.

A simple and precise stability-indicating liquid chromatography method is developed and validated for the quantitative simultaneous estimation of irbesartan (IRB) and hydrochlorothiazide (HCTZ) in combined pharmaceutical dosage form. A chromatographic separation of the two drugs was achieved with an Ace5 C(18) 25-cm analytical column using buffer-acetonitrile (70:30 v/v). The buffer used in mobile phase contains 50 mM ammonium acetate pH adjusted 5.5 with acetic acid. The instrumental settings are flow rate of 1.5 mL/min, column temperature at 30 degrees C, and detector wavelength of 235 nm using a photodiode array detector. IRB, HCTZ, and their combination drug products were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. Peak homogeneity data of IRB and HCTZ is obtained using photodiode array detector. In the stressed sample chromatograms, it demonstrated the specificity of the assay method for their estimation in presence of degradation products. The described method shows excellent linearity over a range of 10-200 microg/mL for IRB and 5-100 microg/mL for HCTZ. Methylparaben was used as internal standard. The correlation coefficient for IRB and HCTZ are 0.998 and 0.999. The mean recovery values for IRB and HCTZ ranged from 100.45% to 101.25%. The limit of detection for IRB and HCTZ were 0.019 and 0.023 microg/mL, respectively, and the limit of quantification were 0.053 and 0.070 microg/mL, respectively. The proposed method was suitable for quantitative determination and stability study of IRB and HCTZ in pharmaceutical preparations and also can be used in the quality control of bulk manufacturing and pharmaceutical dosage forms.

Stability indicating LC method for the simultaneous determination of amlodipine and olmesartan in dosage form.

A simple, rapid, and precise method is developed for the quantitative simultaneous estimation of amlodipine (AM) and olmesartan (OL) in combined pharmaceutical dosage form. A chromatographic separation of the two drugs was achieved with an ACE 5 C(18) 25-cm analytical column using buffer-acetonitrile (60:40, v/v). The resolution between OL and AM was found to be more than 12. Theoretical plates for OL and AM were 6970 and 11,841, respectively. Tailing factor for OL and AM was 0.90 and 0.98, respectively. OL, AM, and combination drug product were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. Peak homogeneity data of OL and AM is obtained by photodiode array detector in the stressed sample chromatograms, demonstrating the specificity of the method for their estimation in presence of degradation product. The described method shows excellent linearity over a range of 20-400 microg/mL for OL and 5-100 microg/mL for AM. The correlation coefficient for OL and AM are 0.9995 and 0.9998, respectively. The relative standard deviation for six measurements in two sets of each drug in tablets is always less than 2%. The proposed method was found to be suitable and accurate for quantitative determination and stability study of OL and AM in pharmaceutical preparations.

Development of GlcNAc-inspired iminocyclitiols as potent and selective N-acetyl-beta-hexosaminidase inhibitors.

Human N-acetyl-beta-hexosaminidase (Hex) isozymes are considered to be important targets for drug discovery. They are directly linked to osteoarthritis because Hex is the predominant glycosidase released by chondrocytes to degrade glycosaminoglycan. Hex is also associated with lysosomal storage disorders. We report the discovery of GlcNAc-type iminocyclitiols as potent and selective Hex inhibitors, likely contributed by the gain of extra electrostatic and hydrophobic interactions. The most potent inhibitor had a K(i) of 0.69 nM against human Hex B and was 2.5 x 10(5) times more selective for Hex B than for a similar human enzyme O-GlcNAcase. These glycosidase inhibitors were shown to modulate intracellular levels of glycolipids, including ganglioside-GM2 and asialoganglioside-GM2.

Analgesic and anti-inflammatory activity of Caryophyllene oxide from Annona squamosa L. bark.

Caryophyllene oxide was isolated from an unsaponified petroleum ether extract of the bark of Annona squamosa and studied for its analgesic and anti-inflammatory activity. Caryophyllene oxide at the doses of 12.5 and 25mg/kg body wt. and unsaponified petroleum ether extract at a dose of 50mg/kg body wt. showed significant central as well as peripheral analgesic, along with anti-inflammatory, activity. These activities of caryophyllene oxide were comparable with the standard drug used in the respective experiments.

Enantiomeric LC separation of valsartan on amylose based stationary phase.

A simple, rapid and robust LC method was developed and validated for the enantiomeric separation of valsartan in bulk drug and formulation. The enantiomers of valsartan were resolved on a Chiralpak AD-H (amylose based stationary phase) column using a mobile phase consisting of n-hexane: 2-propanol: trifluoroacetic acid (85:15:0.2, v/v/v) at a flow rate of 1.0 mL/min. The resolution between the enantiomers was found to be not less than 3.2. The presence of trifluoroacetic acid in the mobile phase played an important role in enhancing chromatographic efficiency and resolution between the enantiomers. The calibration curve for the (R)-enantiomer showed excellent linearity over the concentration range of 600 ng/mL (LOQ) to 6000 ng/mL. The limit of detection and limit of quantification for the (R)-enantiomer were 200 and 600 ng/mL, respectively. The percentage recovery of the (R)-enantiomer ranged between 98.7 to 100.05 % in bulk drug samples of valsartan. The proposed method was found to be suitable and accurate for quantitative determination of (R)-enantiomer in bulk drug substance.

Development and validation of chiral LC method for the enantiomeric separation of duloxetine on amylose based stationary phase.

A simple, rapid, and robust liquid chromatography method was developed and validated for the enantiomeric separation of duloxetine in bulk drug substance. The enantiomers of duloxetine were resolved on a Chiralpak AD-H (amylose based stationary phase) column using a mobile phase consisting of n-hexane-ethanol-diethyl amine (80:20:0.2, v/v/v) at a flow rate of 1.0 mL/min. The resolution between the enantiomers was found to be not less than 2.8 in optimized method. The presence of diethyl amine in the mobile phase played an important role in enhancing chromatographic efficiency and resolution between the enantiomers. The developed method was extensively validated and proved to be robust. The calibration curve for (R)-enantiomer showed excellent linearity over the concentration range of 750 ng/mL (LOQ) to 7500 ng/mL. The limit of detection and quantitation for (R)-enantiomer were 250 and 750 ng/mL, respectively. The percentage recovery of the (R)-enantiomer ranged between 98.3% to 101.05% in bulk drug samples of duloxetine. The proposed method was found to be suitable and accurate for quantitative determination of (R)-enantiomer in bulk drug substance.

Simultaneous high-performance liquid chromatographic determination of telmisartan and hydrochlorothiazide in pharmaceutical preparation.

A simple, rapid, and precise method was developed for the quantitative simultaneous determination of telmisartan and hydrochlorothiazide in combined pharmaceutical dosage form. Chromatographic separation of two drugs was achieved on an ACE 5 C18 25-cm analytical column using buffer-acetonitrile (60:40, v/v) of pH 5.5, adjusted with acetic acid. The buffer used in mobile phase contains 50mM ammonium acetate in double distilled water. The instrumental settings were: flow rate, 1 mL/min; column temperature, 30 degrees C; and detector wavelength, 260 nm. The internal standard method was used for the quantitation of the ingredients of this combination. Methyl paraben was used as an internal standard. The method was validated for linearity, accuracy, precision, limit of detection, limit of quantification, and robustness. The calibration curve shows excellent linearity over the concentration range for telmisartan and hydrochlorothiazide were 10-150 and 5-75 microg/mL, respectively. The correlation coefficient for telmisartan and hydrochlorothiazide were 0.9999. The relative standard deviation for six replicate measurements in two sets of each drug in tablets are always less than 2%. The proposed method was found to be suitable and accurate for quantitative determination of telmisartan and hydrochlorothiazide in pharmaceutical preparation and it can be used for the quality control of formulation products.

Assessment of seasonal variations in surface water quality.

Assessment of seasonal changes in surface water quality is an important aspect for evaluating temporal variations of river pollution due to natural or anthropogenic inputs of point and non-point sources. In this study, surface water quality data for 16 physical and chemical parameters collected from 22 monitoring stations in a river during the years from 1998 to 2001 were analyzed. The principal component analysis technique was employed to evaluate the seasonal correlations of water quality parameters, while the principal factor analysis technique was used to extract the parameters that are most important in assessing seasonal variations of river water quality. Analysis shows that a parameter that is most important in contributing to water quality variation for one season may not be important for another season except for DOC and electrical conductance, which were always the most important parameters in contributing to water quality variations for all four seasons.

Major volatile constituents of Annona squamosa L. bark.

The volatile constituents of Annona squamosa L. bark were identified from the essential oil obtained by steam distillation and studied by GC/MS. Six major components were identified as 1H-Cycloprop(e)azulene (3.46%), germacrene D (11.44%), bisabolene (4.48%), caryophyllene oxide (29.38%), bisabolene epoxide (3.64%) and kaur-16-ene (19.13%). The oil was also screened for its antimicrobial activity, which exhibited a significant antimicrobial activity against Bacillus subtilis and Staphylococcus aureus.