Correction to: Screening and computational analysis of colorectal associated non-synonymous polymorphism in CTNNB1 gene in Pakistani population.

Following publication of the original article [1], the authors have flagged that the article has published with an error in the order of the affiliations.

A novel microfluidic platform for size and deformability based separation and the subsequent molecular characterization of viable circulating tumor cells.

Circulating tumor cells (CTCs) were introduced as biomarkers more than 10 years ago, but capture of viable CTCs at high purity from peripheral blood of cancer patients is still a major technical challenge. Here, we report a novel microfluidic platform designed for marker independent capture of CTCs. The Parsortix™ cell separation system provides size and deformability-based enrichment with automated staining for cell identification, and subsequent recovery (harvesting) of cells from the device. Using the Parsortix™ system, average cell capture inside the device ranged between 42% and 70%. Subsequent harvest of cells from the device ranged between 54% and 69% of cells captured. Most importantly, 99% of the isolated tumor cells were viable after processing in spiking experiments as well as after harvesting from patient samples and still functional for downstream molecular analysis as demonstrated by mRNA characterization and array-based comparative genomic hybridization. Analyzing clinical blood samples from metastatic (n = 20) and nonmetastatic (n = 6) cancer patients in parallel with CellSearch(®) system, we found that there was no statistically significant difference between the quantitative behavior of the two systems in this set of twenty six paired separations. In conclusion, the epitope independent Parsortix™ system enables the isolation of viable CTCs at a very high purity. Using this system, viable tumor cells are easily accessible and ready for molecular and functional analysis. The system's ability for enumeration and molecular characterization of EpCAM-negative CTCs will help to broaden research into the mechanisms of cancer as well as facilitating the use of CTCs as "liquid biopsies."

Maternal and foetal outcome of 206 high risk pregnancy cases in border guard hospital, dhaka.

This observational study was carried out to identify the various types of high risk pregnancy and to determine the maternal and foetal outcome. The study was carried out on 206 pregnant high risk women in the Gynecology and Obstetrics department of Border Guard Hospital, Dhaka from January 2012 to December 2012. During mentioned period among 598 pregnant women 206 high risk pregnancy cases were randomly selected. Pregnant women (gestational age from 34 weeks upto 40 weeks) having medical condition and pregnancy related high risk factors were included and uncomplicated pregnancy, pregnancy before 37 weeks, post dated pregnancy were excluded from this study. Data was collected from semi structured history sheet and data analysis done by percentage. High risk pregnant women were grouped into three. Group A and Group B includes pregnant women having medical condition before and during pregnancy respectively. Group C consists of pregnant women had pregnancy related high risk issues. Among 206 high risk pregnancy cases majority 47.57% women had medical condition during pregnancy, 31.55% patient had medical condition before pregnancy. Among them majority 30.58% of the patient suffered from pregnancy induced hypertension, 15.04% patients suffered from gestational Diabetes Mellitus and premature rupture of membranes were 12.13%. In this study majority 43.68% of high risk pregnant patients were in age group of 30-35 years, 19.90% pregnant women were in age group of >35 years and 19.40% were in age group of upto 20 years. Among study groups maximum 65.04% of the patients were multiparous. Among 206 study population 60.19% high risk pregnant women were at term at the time of delivery and 39.8% women delivered their babies preterm. Caesarean section was done in 69.41% of high risk pregnant women. After delivery majority 77.66% women had no complication, only 10.19%, 8.25%, 2.91% and 0.97% high risk pregnant women suffered from fever, UTI, abdominal wound infection and post partum hemorrhage respectively. In this study, among 206 pregnancy cases 91.31% of the neonates had Apgar score >7 and 8.61% neonates had Apgar score <7%, 33.49% neonates had low birth weight and premature 39.80%. During the study period no maternal and neonatal death were observed.

Composition and Antimicrobial Activity of the Essential Oil from Leaves of Curcuma longa L. Kasur Variety.

The essential oil from the leaves of Curcuma longa L. Kasur variety grown in Pakistan was extracted by hydro-distillation. Chemical constituents of the essential oil were identified by gas chromatography/mass spectrometry. The chromatographic analysis of oil showed 25 constituents, out of which nine chemical constituents were identified. The eucalyptol (10.27%) was the major component of the essential oil. α-pinene (1.50%), ß-phellandrene (2.49%), ß-pinene (3.57%), limonene (2.73%), 1,3,8-p-menthatriene (1.76%), ascaridole epoxide (1.452%), 2-methylisoborneol (2.92%), 5-isopropyl-6-methyl-hepta-3, dien-2-ol (2.07%) were also present in considerable quantity. The antimicrobial properties of leaves of Curcuma longa were tested by disc diffusion method against various human pathogens, including eight fungal and five bacterial strains. Essential oil showed maximum resistance against Fusarium miniformes MAY 3629 followed by Bacillus subtilis ATCC 6633 whereas; it exhibited least resistance against Fusarium oxysporium ATCC 48122. The results of the antimicrobial assay revealed that essential oil showed significant inhibitory activity against the tested organisms.

Prevalent genotypes of meticillin-resistant Staphylococcus aureus: report from Pakistan.

Meticillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen in Pakistan and is emerging in the community. This is one of the first reports of the prevalent genotypes of MRSA in both hospital and community settings in Pakistan. Isolates collected in 2006-2007 were characterized by PFGE, staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). PFGE identified nine pulsotypes, the majority of isolates belonging to pulsotypes A (n=70) and B (n=38), which were predominant among hospital-onset MRSA (HO-MRSA) and community-onset MRSA (CO-MRSA) isolates, respectively. Among the HO-MRSA isolates, variants of SCCmec type III were prevalent, whilst SCCmec type IV or variants were predominant in the CO-MRSA isolates. MLST identified two principal sequence types, ST8 and ST239. An association was observed between ST8, PFGE pulsotype B and SCCmec type IV in the CO-MRSA (ST8-MRSA-IV). Similarly, ST239, PFGE pulsotype A and SCCmec type III were associated with HO-MRSA (ST239-MRSA-III). Therefore, the prevalent genotypes circulating in Pakistan at the time of study were ST8-MRSA-IV and ST239-MRSA-III in the community and hospital settings, respectively. A set of HO-MRSA isolates collected in 1997 were characterized by PFGE and SCCmec typing for comparison. The isolates belonged to two PFGE pulsotypes (A, n=28; B, n=11) and contained just two SCCmec types. These results suggest that an increase in genetic diversity occurred over the period 1997-2007 as a result of either microevolution or the importation of strains from surrounding areas.

Pattern of police torture in Punjab, Pakistan.

A total of 1820 victims of alleged police torture were examined at the office of Surgeon Medicolegal Punjab Lahore during a period of 5 years. Most of the victims at the time of examination were showing visible evidence of Physical trauma. Victims were mainly men. Examination was conducted on the directions of various courts (Judicial Magistrates, District and Session Judges, and Lahore High Court). A wide range of different types of injuries of different durations were observed on various parts of the body. Blunt trauma was most frequent. Psychologic element of torture was also seen in some victims.

Birth weight percentiles by gestational age: a hospital based study.

BACKGROUND: There are no present or old large population-based birth weight normograms available for Pakistani population. The aim of the study was to develop birth weight for gestational age normograms for singleton and twin births, based on perinatal data collected prospectively. METHODS: Birth weight percentiles by gestational age were determined in women delivering at Agha Khan University Hospital Karachi from January 1992 to December 1994. This was a prospective cohort study and included all women who delivered from 28 weeks to 44 weeks of gestation. Singleton as well as twin pregnancies were included but women having intrauterine or intrapartum deaths were excluded. RESULTS: Data of 4041 live births were collected from 4041 deliveries. Total infants were 4112, there being 71 sets of twins. The male babies were heavier than female babies at each gestational week. There was a preponderance of male infants for each week. Out of 4041 deliveries, 414 babies were born before 37 completed weeks; percentage of preterm birth is 10.24%. Males were more likely to be born preterm then were females, although females were more likely to be of low birth weight. Out of 414 preterm births, 281 were male and 183 were female. Out of them data of 4030 live births was compiled and analysed; percentiles were formed and compared with other studies.

D-Aspartate oxidase and D-amino acid oxidase are localised in the peroxisomes of terrestrial gastropods.

D-Aspartate oxidase and D-amino acid oxidase were found in high activity in the tissues of representative species of terrestrial gastropods. Analytical subcellular fractionation demonstrated that both of these oxidases co-localised with the peroxisome markers, acyl-CoA oxidase and catalase, in the digestive gland homogenate. Electron microscopy of peak peroxisome fractions showed particles of uniform size with generally well preserved variably electron-dense matrices bounded by an apparently single limiting membrane. Many of the particles exhibited a core region of enhanced electron density. Catalase cytochemistry of peak fractions confirmed the peroxisome identity of the organelles. Peroxisome-enriched subcellular fractions were used to investigate the properties of gastropod D-aspartate oxidase and D-amino acid oxidase activities. The substrate and inhibitor specificities of the two activities demonstrated that two distinct enzymes were present analogous to, but not identical to, the equivalent mammalian peroxisomal enzymes.

Stability of organophosphate and pyrethroid pesticides on wheat in storage.

The pesticides chlorpyriphos-methyl, pirimiphos-methyl and permethrin were applied to wheat and stored for 52 weeks at 25, 30, 35 and 40 degrees C, and at 10 and 13% m.c. Rates of loss were calculated from the residue analyses of pesticides in treated wheat at monthly intervals during the storage period. Calculated half-lives and pseudo first-order rate constants of these pesticides are discussed with reference to temperature and moisture.

Gastropod mollusc aliphatic alcohol oxidase: subcellular localisation and properties.

The digestive gland and other tissues of several species of terrestrial gastropod mollusc contain an aliphatic alcohol oxidase activity (EC1.1.3.13). The enzyme is FAD dependent, consumes oxygen and generates hydrogen peroxide and the corresponding aldehyde. Saturated primary alcohols are favoured as substrates with octanol preferred with an apparent Km of 3-4 microM. The activity is clearly distinguishable from previously reported molluscan aromatic alcohol oxidase (EC1.1.3.7) on the basis of FAD dependence, sensitivity to heat treatment and high salt concentration and with regard to substrate preferences. The aliphatic alcohol oxidase is membrane associated and most likely localised to the endoplasmic reticulum. Extraction of membranes with 1% Igipal solubilises the enzyme in active form. This enzyme is a further example of an oxidase apparently restricted to molluscs.

Spleen necrosis virus-derived C-type retroviral vectors for gene transfer to quiescent cells.

Gene therapy applications of retroviral vectors derived from C-type retroviruses have been limited to introducing genes into dividing target cells. Here, we report genetically engineered C-type retroviral vectors derived from spleen necrosis virus (SNV), which are capable of infecting nondividing cells. This has been achieved by introducing a nuclear localization signal (NLS) sequence into the matrix protein (MA) of SNV by site-directed mutagenesis. This increased the efficiency of infecting nondividing cells and was sufficient to endow the virus with the capability to efficiently infect growth-arrested human T lymphocytes and quiescent primary monocyte-derived macrophages. We demonstrate that this vector actively penetrates the nucleus of a target cell, and has potential use as a gene therapy vector to transfer genes into nondividing cells.

Technology evaluation: PRO-542, Progenics Pharmaceuticals inc.

Progenics's rCD4-IgG2 (PRO-542) is a recombinant fusion protein, which has been developed using the company's Universal Antiviral Binding (UnAB) technology, and is in phase I/II clinical trials for the treatment of human immunodeficiency virus type I (HIV-1) infection [273391]. At the beginning of 1997, Progenics received a Phase II Small Business Innovation Research Program (SBIR) grant from the National Institute of Allergy and Infectious diseases (NIAID) to fund the development of PRO-542 [236048]. A further grant of $2.7 million was awarded in August 1998 for the clinical evaluation of PRO-542 and other anti-HIV therapies [294200]. Progenics is collaborating with the Aaron Diamond AIDS Research Center (ADARC) in New York and the Center for Disease Control and Prevention in Atlanta [178410]. In February 2000, Progenics and Genzyme Transgenics Corp signed an agreement to continue the development of a transgenic source of PRO-542. Genzyme will develop transgenic goats that produce PRO-542 in their milk in exchange for undisclosed fees and milestone payments. Genzyme will supply PRO-542 to Progenics for clinical trials with a possibility for eventual commercial supply [357291]. Following on from this, in October 2000, Progenics received an SBIR grant to fund a two-year project with Genzyme Transgenics into the development of cost-effective methods for the manufacture of PRO-542, by optimization of the production of the drug in the milk of transgenic dairy animals [385982]. In August 2000, Punk, Ziegel & Company predicted that Progenics Pharmaceuticals will become sustainably profitable in 2003 following the launch of PRO-542 and GMK (Progenics Pharmaceuticals) in 2002 [390063].

Antibacterial steroidal alkaloids from Sarcococca saligna.

Two new pregnane-type steroidal alkaloids, saligcinnamide [(20S,2'E)-20-(N,N-dimethylamino)-3beta-(3'-phenyl-2'-propenyl-N-meth ylamido)pregnane](1) and N(a)-methyl epipachysamine-D [(20S)-20-(N,N-dimethylamino)-3beta-(N-methylbenzamido)pregnane](2 ), along with a known base, epipachysamine D [(20S)-20-(N,N-dimethylamino)-3beta-(benzamido)pregnane] (3), were isolated from the EtOH extracts of the roots and stems of Sarcococca saligna. The new bases exhibited antibacterial activity against several human pathogenic bacteria. Two derivatives of 1, dihydrosaligcinnarnide [(20S)-20-(N,N-dimethylamino)-3beta-(3'-phenylpropionoyl-N-meth ylamido)pregnane](4) and dihydrosaligcinnamine [(20S)-20-(N,N-dimethylamino)-3beta-N-(3'-phenylpropyl-N-methylamino)pre gnane](5), and a derivative of 2, N(a)-methyl epipachysamine [(20S)-20-(N,N-dimethylamino)-3beta-(N-benzyl,N-methylamino)pregnane](6) were prepared and their antibacterial activity determined.

New steroidal alkaloids from Buxus longifolia.

Four new steroidal alkaloids, (+)-cyclovirobuxeine F (1), N-benzoyl-O-acetylbuxalongifoline (2), buxasamarine (3), and (+)-cyclobuxamidine (4), along with two known steroidal bases, 16 alpha-acetoxybuxabenzamidienine (5) and trans-cyclosuffrobuxinine (6), were isolated from the leaves of Buxus longifolia. The alkaloids 1-4 showed significant antibacterial activity.