The structural characterization and bioactivity assessment of nonspecific lipid transfer protein 1 (nsLTP1) from caraway (Carum carvi) seeds.

BACKGROUND: Carum carvi (caraway) of the Apiaceae family has been used in many cultures as a cooking spice and part of the folk medicine. Previous reports primarily focus on the medicinal properties of caraway seed essential oil and the whole seeds extract. However, no effort has been made to study caraway proteins and their potential pharmacological properties, including nonspecific lipid transfer protein (nsLTP), necessitating further research. The current study aimed to characterize nonspecific lipid transfer protein 1 (nsLTP1) from caraway seed, determine its three-dimensional structure, and analyze protein-ligand complex interactions through docking studies. We also evaluated nsLTP1 in vitro cytotoxic effect and antioxidant capacity. Additionally, nsLTP1 thermal- and pH- stability were investigated. METHODS: Caraway nsLTP1 was purified using two-dimensional chromatography. The complete amino acid sequence of nsLTP1 was achieved by intact protein sequence for the first 20 residues and the overlapping digested peptides. The three-dimensional structure was predicted using MODELLER. Autodock Vina software was employed for docking fatty acids against caraway nsLTP1. Assessment of nsLTP1 cytotoxic activity was achieved by MTS assay, and the Trolox equivalent antioxidant capacity (TAC) was determined. Thermal and pH stability of the nsLTP1 was examined by circular dichroism (CD) spectroscopy. RESULTS: Caraway nsLTP1 is composed of 91 residues and weighs 9652 Da. The three-dimensional structure of caraway nsLTP1 sequence was constructed based on searching known structures in the PDB. We chose nsLTP of Solanum melongena (PDB ID: 5TVI) as the modeling template with the highest identity among all other homologous proteins. Docking linolenic acid with caraway protein showed a maximum binding score of -3.6 kcal/mol. A preliminary screening of caraway nsLTP1 suppressed the proliferation of human breast cancer cell lines MDA-MB-231 and MCF-7 in a dose­dependent manner with an IC50 value of 52.93 and 44.76 µM, respectively. Also, nsLTP1 (41.4 µM) showed TAC up to 750.4 µM Trolox equivalent. Assessment of nsLTP1 demonstrated high thermal/pH stability. CONCLUSION: To the best of our knowledge, this is the first study carried out on nsLTP1 from caraway seeds. We hereby report the sequence of nsLTP1 from caraway seeds and its possible interaction with respective fatty acids using in silico approach. Our data indicated that the protein had anticancer and antioxidant activities and was thermally stable.

Aflatoxin M1 in fresh milk collected from local markets of Karachi, Pakistan.

During 2016-2017, 156 samples of fresh milk samples were collected from local markets of Karachi, Pakistan and analysed for aflatoxin M1 (AFM1) contamination using ELISA technique. AFM1 was detected in 143 (91.7%) samples, ranged from 20 to 3090 ng L-1 with a mean level of 346.2 ng L-1. In 125 (80.1%) samples, the AFM1 contamination was greater than the maximum limit (ML = 50 ng L-1) set by EU. However, in 51 (32.7%) samples, the AFM1 level was higher than the ML of 500 ng L-1 as assigned by the USA. Statistical analysis showed that the AFM1 level in milk samples from summer was significantly (p < 0.05) higher than that obtained in winter. It was concluded that the AFM1 levels in the tested samples appear to be a serious public health problem. Therefore, immediate measures should be taken and re-evaluation done for the procedures for farming, transportation, refrigeration, and storage for the control of AFM1 level in milk samples.

Engaging with community-based public and private mid-level providers for promoting the use of modern contraceptive methods in rural Pakistan: results from two innovative birth spacing interventions.

BACKGROUND: Family planning (FP) interventions aimed at reducing population growth have negligible during the last two decades in Pakistan. Innovative FP interventions that help reduce the growing population burden are the need of the hour. Marie Stopes Society--Pakistan implemented an operational research project--'Evidence for Innovating to Save Lives', to explore effective and viable intervention models that can promote healthy timing and spacing of pregnancy in rural and under-served communities of Sindh, Punjab and Khyber Pakhtunkhwa provinces of Pakistan. METHODS: We conducted a quasi-experimental (pre- and post-intervention with control arm) study to assess the effectiveness of each of the two intervention models, (1) Suraj model (meaning 'Sun' in English), which uses social franchises (SF) along with a demand-side financing (DSF) approach using free vouchers, and (2) Community Midwife (CMW) model, in promoting the use of modern contraceptive methods compared to respective controls. Baseline and endline cross-sectional household surveys were conducted, 24 months apart, by recruiting 5566 and 6316 married women of reproductive age (MWRA) respectively. We used Stata version 8 to report the net effect of interventions on outcome indicators using difference-in-differences analysis. Multivariate Cox proportional hazard regression analysis was used to assess the net effect of the intervention on current contraceptive use, keeping time constant and adjusting for other variables in the model. RESULTS: The Suraj model was effective in significantly increasing awareness about FP methods among MWRA by 14% percentage points, current contraceptive use by 5% percentage points and long term modern method--intrauterine device (IUD) use by 6% percentage points. The CMW model significantly increased contraceptive awareness by 28% percentage points, ever use of contraceptives by 7% percentage points and, IUD use by 3% percentage points. Additionally the Suraj intervention led to a 35% greater prevalence (prevalence ratio: 1.35, 95% CI: 1.22-1.50) of contraceptive use among MWRA. CONCLUSION: Suraj intervention highlights the importance of embedding subsidized FP services within the communities of the beneficiaries. The outcomes of the CMW intervention also improved the use of long-term contraceptives. These findings indicate the necessity of designing and implementing FP initiatives involving local mid-level providers to expand contraceptive coverage in under-served areas.

Family Planning Knowledge, Attitudes, and Practices among Married Men and Women in Rural Areas of Pakistan: Findings from a Qualitative Need Assessment Study.

This paper presents the findings of a qualitative assessment aimed at exploring knowledge, attitudes, and practices regarding family planning and factors that influence the need for and use of modern contraceptives. A descriptive exploratory study was conducted with married women and men aged between 15 and 40. Overall, 24 focus group discussions were conducted with male and female participants in three provinces of Pakistan. The findings reveal that the majority knew about some modern contraceptive methods, but the overall contraceptive use was very low. Knowledge and use of any contraceptive method were particularly low. Reasons for not using family planning and modern contraception included incomplete family size, negative perceptions, in-laws' disapproval, religious concerns, side-effects, and lack of access to quality services. The majority preferred private facilities over the government health facilities as the later were cited as derided. The study concluded the need for qualified female healthcare providers, especially for long term family planning services at health facilities instead of camps arranged occasionally. Addressing issues around access, affordability, availability, and sociocultural barriers about modern contraception as well as involving men will help to meet the needs and ensure that the women and couples fulfill their childbearing and reproductive health goals.

Aflatoxins contamination in Pakistani brown rice: a comparison of TLC, HPLC, LC-MS/MS and ELISA techniques.

Advancement in the field of analytical food-chemistry has explored various experimental techniques for aflatoxins (AFs) quantification. The present study was aimed to compare four different techniques; thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC-MS/MS) and enzyme-linked immunosorbent assay (ELISA) for the analysis of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in brown rice (n = 120) being collected from Karachi, Pakistan. All the four assays provide precised, accurate and comparable results. However, some differences were observed. For instance, TLC, HPLC and LC-MS/MS methodologies offered the advantage of the quantification of individual toxins in contrast to ELISA technique. The contamination ranges of AFB1/AFB2 as determined by TLC, HPLC and LC-MS/MS were 1.18-9.97/0.59-1.52, 0.16-10.54/0.26-1.35 and 0.11-10.88/0.38-1.48 µg/kg, respectively. However, AFG1 and AFG2 were not detected in any tested samples. Furthermore, owing to low-detection limit and sensitivity, HPLC and LC-MS/MS methodologies have identified greater number of contaminated samples in comparison to TLC and ELISA techniques. The overall average results of total AFs as provided by HPLC (3.79 µg/kg) and LC-MS/MS (3.89 µg/kg) were found higher in comparison to TLC (3.68 µg/kg) and ELISA (3.70 µg/kg). On the basis of achieved results, it was concluded that TLC, HPLC, LC-MS/MS and ELISA techniques are valuable tool for the quantification of AFs in cereals and grains. Furthermore, HPLC and LC-MS/MS techniques offer an added advantage for the detection of AFs in diminutive levels.

Impact of social franchising on contraceptive use when complemented by vouchers: a quasi-experimental study in rural Pakistan.

BACKGROUND: Pakistan has had a low contraceptive prevalence rate for the last two decades; with preference for natural birth spacing methods and condoms. Family planning services offered by the public sector have never fulfilled the demand for contraception, particularly in rural areas. In the private sector, cost is a major constraint. In 2008, Marie Stopes Society - a local NGO started a social franchise programme along with a free voucher scheme to promote uptake of IUCDs amongst the poor. This paper evaluates the effectiveness of this approach, which is designed to increase modern long term contraceptive awareness and use in rural areas of Pakistan. METHODOLOGY: We used a quasi-experimental study design with controls, selecting one intervention district and one control district from the Sindh and Punjab provinces. In each district, we chose a total of four service providers. A baseline survey was carried out among 4,992 married women of reproductive age (MWRA) in February 2009. Eighteen months after the start of intervention, an independent endline survey was conducted among 4,003 women. We used multilevel logistic regression for analysis using Stata 11. RESULTS: Social franchising used alongside free vouchers for long term contraceptive choices significantly increased the awareness of modern contraception. Awareness increased by 5% in the intervention district. Similarly, the ever use of modern contraceptive increased by 28.5%, and the overall contraceptive prevalence rate increased by 19.6%. A significant change (11.1%) was recorded in the uptake of IUCDs, which were being promoted with vouchers. CONCLUSION: Family planning franchise model promotes awareness and uptake of contraceptives. Moreover, supplemented with vouchers, it may enhance the use of IUCDs, which have a significant cost attached. Our research also supports a multi-pronged approach- generating demand through counselling, overcoming financial constraints by offering vouchers, training, accreditation and branding of the service providers, and ensuring uninterrupted contraceptive supplies.

Assessment of safety and efficacy of arterial embolisation in the management of intractable epistaxis.

To evaluate outcomes of intractable epistaxis managed with arterial embolisation. Fourteen sequential cases of intractable epistaxis that underwent embolisation in our centre were evaluated retrospectively and interviewed over the phone. All patients had several failed treatment modalities prior to embolisation. Patients' follow up ranged from 1 to 57 months with median of 26 months. All 14 cases underwent a single embolisation procedure with successful arrest of epistaxis. Four cases (29%) developed recurrent epistaxis at a later date. One (7%) required re-embolisation 19 months after his first procedure. One bled 17 days after embolisation, but this settled with hospital admission and Bismuth Iodoform Paraffin Paste packing. The other two developed minor episodes of epistaxis, which did not require hospital admission. Two patients developed local ischaemic complications following arterial embolisation. Of those, one developed necrosis of the left alar skin and cartilage that healed reasonably well after 5 months. The other case developed mucosal necrosis of the right side of the hard palate; this patient was the one who bled 17 days post-embolisation. The palatal necrosis healed in a satisfactory manner without causing any functional impairment of the oral cavity. Embolisation is a successful intervention in management of persistent epistaxis, when other interventions fail. The risks of major complications such as stroke are well known, and discussed with patients prior to the procedure. It is also important to discuss the risks of ischaemic damage to the face and oral cavity. In our experience, these complications have been minor and the benefits still outweigh the complications.

The "doctor-customer" relationship: Hippocrates in the modern marketplace.

We performed a consecutive survey of 100 people presenting to a hospital injury clinic to ascertain their attitude to terminology currently used to describe them in our own institution and in the international literature. The results of this demonstrated that the subjects significantly preferred the traditional assignation "patient" rather than terms such as client or customer. This finding reflects the need to remember peoples' attitudes and expectations from their consultation with their doctor. Although business models undoubtedly help in the provision of an efficient health care service, remaining at the centre of this encounter is a doctor-patient relationship that involves a more complex interaction than simply a market transaction.

Simultaneous hip and upper limb fracture in the elderly: incidence, features and management considerations.

We performed a retrospective analysis of all patients admitted to a single unit over 4 years with fractured neck of femur. Of the 760 patients admitted over this period of time 36 (4.7%) were found to also have a fracture of the upper limb. The associated upper limb fractures were distal radius (n=28), olecranon (n=2) and neck of humerus (n=5) and fifth metacarpal (n=1), with the same ratio of intracapsular to extracapsular fractures as the whole group. The female to male ratio in the isolated hip fracture group was 2.4:1 and for the combined fractures group was 8:1 (difference P=0.014). The mean patient age was 77.3 years for isolated hip fractures and 83.9 for the combined group (P=0.037). The mean total length of stay in hospital for isolated hip fracture was 15.6 days and for combined fractures was 20.4 days (P=0.010). We have demonstrated that combined upper limb and neck of femur fractures occur in a population that is older and predominantly female. They are associated with a significant increase in hospital stay and increased difficulties in mobilisation as a consequence of the combined fractures. It is therefore important to recognise this specific subgroup of patients presenting with hip fractures in order to ensure that they receive adequate treatment and rehabilitation and that the use of valuable health care resources are optimised.