High-throughput genotyping of high-homology mutant mouse strains by next-generation sequencing.

Genotyping of knockout alleles in mice is commonly performed by end-point PCR or gene-specific/universal cassette qPCR. Both have advantages and limitations in terms of assay design and interpretation of results. As an alternative method for high-throughput genotyping, we investigated next generation sequencing (NGS) of PCR amplicons, with a focus on CRISPR-mediated exon deletions where antibiotic selection markers are not present. By multiplexing the wild type and mutant-specific PCR reactions, the genotype can be called by the relative sequence counts of each product. The system is highly scalable and can be applied to a variety of different allele types, including those produced by the International Mouse Phenotyping Consortium and associated projects. One potential challenge with any assay design is locating unique areas of the genome, especially when working with gene families or regions of high homology. These can result in misleading or ambiguous genotypes for either qPCR or end-point assays. Here, we show that genotyping by NGS can negate these issues by simple, automated filtering of undesired sequences. Analysis and genotype calls can also be fully automated, using FASTQ or FASTA input files and an in-house Perl script and SQL database.

Elucidation of larvicidal potential of metallic and environment friendly food-grade nanostructures against Aedes albopictus.

To combat health challenges associated with mosquito-borne diseases, the larvicidal activity of metallic nanoparticles, food-grade polymeric nano-capsules and insecticides was investigated against larvae of Aedes albopictus as an effective alternate control approach. The Ae. albopictus was identified using sequencing and phylogenetic analyses of COXI, CYTB and ITS2 genes. The characterization of synthesized nanostructures was performed through Zetasizer, UV-VIS spectroscopy, atomic force microscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy. The mosquito larvae were exposed to varying concentration of nanostructures and insecticides, and their percentage mortality was evaluated at different time intervals of 24 h and 48 h exposure. The highest efficacy was observed in zinc oxide nanoparticles (ZnO-NPs) and polymeric nanocapsules FG-Cur E-III (LC50 = 0.24 mg/L, LC90 = 0.6 mg/L) and (LC50 = 3.8 mg/L, LC90 = 9.33 mg/L), respectively, after 24 h; while (LC50 = 0.18 mg/L, LC90 = 0.43 mg/L) and (LC50 = 1.95 mg/L, LC90 = 6.46 mg/L), respectively, after 48 h against fourth instar larvae of Ae. albopictus. Ag, CuO, NiTiO3 and CoTiO3 nanoparticles evaluated in this study also showed promising larvicidal activity. Although ZnO-NPs proved to be effective larvicides, their possible toxicity (producing ROS species) can limit their use. The curcumin nanostructures (FG-Cur E-III) stabilized by food-grade materials are thought to exert their larvicidal activity by binding to sterol carrier protein-2, and depriving the larvae from the essential dietary cholesterol, and bears effective larvicidal potential as safe alternative for chemical larvicides, due to their environment friendly, food-grade and easy biodegradability.

The HAPPY (Healthy and Active Parenting Programmme for early Years) feasibility randomised control trial: acceptability and feasibility of an intervention to reduce infant obesity.

BACKGROUND: The prevalence of infant obesity is increasing, but there is a lack of evidence-based approaches to prevent obesity at this age. This study tested the acceptability and feasibility of evaluating a theory-based intervention aimed at reducing risk of obesity in infants of overweight/obese women during and after pregnancy: the Healthy and Active Parenting Programme for Early Years (HAPPY). METHODS: A feasibility randomised controlled trial was conducted in Bradford, England. One hundred twenty overweight/obese pregnant women (Body Mass Index [BMI] ≥25 kg/m(2)) were recruited between 10-26 weeks gestation. Consenting women were randomly allocated to HAPPY (6 antenatal, 6 postnatal sessions: N = 59) or usual care (N = 61). Appropriate outcome measures for a full trial were explored, including: infant's length and weight, woman's BMI, physical activity and dietary intake of the women and infants. Health economic data were collected. Measurement occurred before randomisation and when the infant was aged 6 months and 12 months. Feasibility outcomes were: recruitment/attrition rates, and acceptability of: randomisation, measurement, and intervention. Intra-class correlations for infant weight were calculated. Fidelity was assessed through observations and facilitator feedback. Focus groups and semi-structured interviews explored acceptability of methods, implementation, and intervention content. RESULTS: Recruitment targets were met (~20 women/month) with a recruitment rate of 30 % of eligible women (120/396). There was 30 % attrition at 12 months; 66 % of recruited women failed to attend intervention sessions, but those who attended the first session were likely to continue to attend (mean 9.4/12 sessions, range 1-12). Reaction to intervention content was positive, and fidelity was high. Group clustering was minimal; an adjusted effect size of -0.25 standard deviation scores for infant weight at 12 months (95 % CI: -0.16-0.65) favouring the intervention was observed using intention to treat analyses. No adverse events were reported. CONCLUSIONS: The HAPPY intervention appeared feasible and acceptable to participants who attended and those delivering it, however attendance was low; adaptations to increase initial attendance are recommended. Whilst the study was not powered to detect a definitive effect, our results suggest a potential to reduce risk of infant obesity. The evidence reported provides valuable lessons to inform progression to a definitive trial. TRIAL REGISTRATION: Current Controlled Trials ISRCTN56735429.

Overexpression of the Hspa13 (Stch) gene reduces prion disease incubation time in mice.

Prion diseases are fatal neurodegenerative disorders that include bovine spongiform encephalopathy (BSE) and scrapie in animals and Creutzfeldt-Jakob disease (CJD) in humans. They are characterized by long incubation periods, variation in which is determined by many factors including genetic background. In some cases it is possible that incubation time may be directly correlated to the level of gene expression. To test this hypothesis, we combined incubation time data from five different inbred lines of mice with quantitative gene expression profiling in normal brains and identified five genes with expression levels that correlate with incubation time. One of these genes, Hspa13 (Stch), is a member of the Hsp70 family of ATPase heat shock proteins, which have been previously implicated in prion propagation. To test whether Hspa13 plays a causal role in determining the incubation period, we tested two overexpressing mouse models. The Tc1 human chromosome 21 (Hsa21) transchromosomic mouse model of Down syndrome is trisomic for many Hsa21 genes including Hspa13 and following Chandler/Rocky Mountain Laboratory (RML) prion inoculation, shows a 4% reduction in incubation time. Furthermore, a transgenic model with eightfold overexpression of mouse Hspa13 exhibited highly significant reductions in incubation time of 16, 15, and 7% following infection with Chandler/RML, ME7, and MRC2 prion strains, respectively. These data further implicate Hsp70-like molecular chaperones in protein misfolding disorders such as prion disease.

Microglial Cx3cr1 knockout reduces prion disease incubation time in mice.

BACKGROUND: Microglia are resident mononuclear phagocytes of the brain that become activated in response to insults including neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and prion disease. In the central nervous system the chemokine Cx3cl1 (Fractalkine) is expressed by neurons and its exclusive receptor Cx3cr1 is expressed solely on microglia. Cx3cl1/Cx3cr1 signalling is thought to maintain microglia in their resting state and disrupting this equilibrium may allow microglia to become activated. In prion disease, microglial proliferation has been suggested to contribute to overall disease progression, however, in different mouse models of neurodegeneration, loss of Cx3cr1 has been shown to either worsen or improve the phenotype depending on the paradigm. RESULTS: To investigate the role of Cx3cl1/Cx3cr1 signalling in prion disease we infected Cx3cr1 null mice with three different strains of prions. Following challenge with Chandler/RML, ME7 and MRC2 prion strains, Cx3cr1 knockout mice showed highly significant reductions in incubation time. No differences were seen in the pattern and localisation of activated microglia in the brain or in the mRNA expression levels of chemokines/cytokines (Cxcl10, Il-12b, Il-1b, Arg-1 and Cxc3l1). CONCLUSION: Our data suggest a protective role for Cx3cl1/Cx3cr1 cross-talk in prion disease.

Personal protective equipment and micro-nano plastics: A review of an unavoidable interrelation for a global well-being hazard.

The usage and the demand for personal protective equipments (PPEs) for our day-to-day survival in this pandemic period of COVID-19 have seen a steep rise which has consequently led to improper disposal and littering. Fragmentation of these PPE units has eventually given way to micro-nano plastics (MNPs) emission in the various environmental matrices and exposure of living organisms to these MNPs has proven to be severely toxic. Numerous factors contribute to the toxicity imparted by these MNPs that mainly include their shape, size, functional groups and their chemical diversity. Even though multiple studies on the impacts of MNPs toxicity are available for other organisms, human cell line studies for various plastic polymers, other than the most common ones namely polyethylene (PE), polystyrene (PS) and polypropylene (PP), are still at their nascent stage and need to be explored more. In this article, we cover a concise review of the literature on the impact of these MNPs in biotic and human systems focusing on the constituents of the PPE units and the additives that are essentially used for their manufacturing. This review will subsequently identify the need to gather scientific evidence at the smaller level to help combat this microplastic pollution and induce a more in-depth understanding of its adverse effect on our existence.

The commitment to a midwifery centre care model in Bangladesh: An interview study with midwives, educators and students.

BACKGROUND: Midwifery-led care is a key factor in reducing maternal and new-born mortality globally. In Bangladesh, only a third of births are attended by professionals and almost 70% of births occur outside healthcare facilities. Midwifery is a relatively new profession in Bangladesh and a midwifery centre care model has only recently been introduced. This study aims to explore the willingness within the healthcare system to support a greater role for midwifery centres in maternity services. METHODS: Data were collected through individual semi-structured interviews with 55 midwives, midwifery educators and final year midwifery students. Two of the midwifery educators were principals of nursing institutes involved in the government's midwifery leadership and considered as experts in the midwifery care system. The data was analysed using qualitative content analysis. The transcribed interviews comprised 150 pages. The study received ethical approval from the Directorate General of Nursing and Midwifery in Bangladesh. RESULTS: One main category emerged from the study: "The foundations of a midwifery centre care model need to be strengthened for the sustainable implementation of midwifery centres in Bangladesh to continue". Five additional categories were identified: 1) The midwifery centre care model is inaccessible for communities, 2) Striving for acceptable standards of care within a midwifery centre care model is not a priority 3) Respectful, woman-centred care is weak, 4) Community engagement with the midwifery centre care model is insufficient, and 5) The midwifery centre care model is not integrated into the healthcare system. These categories were supported by the identification of 11 sub-categories. CONCLUSION: The willingness to commit to a midwifery centre care model is not yet in place in Bangladesh. Advocacy, information, and education about the benefits of normal birth assisted by professional midwives is needed at all levels of Bangladeshi society.

Efficacy of itraconazole versus fluconazole in vaginal candidiasis.

OBJECTIVE: To compare the efficacy of fluconazole 150mg single dose and itraconazole 200mg twice for one day in the treatment of acute vulvovaginal candidiasis. METHODS: The study was carried out at the Department of Dermatology, PNS Shifa Hospital, Karachi, from March, 2008 to February 2009 and comprised 60 women with clinical and mycological diagnosis of vaginal candidiasis. Diagnosis was based on history, clinical examination and relevant investigations. The women were divided into two equal groups. After initial assessment, Group 1 was treated with capsule fluconazole 150mg stat, and Group 2 with capsule itraconazole 200mg twice for one day. They were assessed clinically for cure and relapse on day 7 and 21 respectively. All findings were recorded in the proforma. Data was analysed using SPSS 12. RESULTS: The overall clinical evaluation showed 70% (n = 21) cure rate with itraconazole and 50% (n = 15) with fluconazole. In Group-1, 7 (23.33%) and in Group-2 8 (26.6%) showed some improvement, while 2 (6.66%) in Group 1, and 7 (23%) in Group 2 failed to respond. Relapse was observed in 9 (28.5%) and 16 (53%) of the cured cases in Group 1 and Group 2 respectively. CONCLUSION: Itraconazole was found to be more effective in the treatment of vulvovaginal candidiasis compared to fluconazole with high cure and low relapse rate.

Frequency and clinical variants of specific dermatoses in third trimester of pregnancy: a study from a tertiary care centre.

OBJECTIVE: To determine the frequency of patients with dermatoses in the third trimester of pregnancy and to identify various clinical types of dermatoses in the third trimester. METHODS: The study was carried out at the Department of Dermatology and the Department of Gynaecology & Obstetrics, PNS Shifa Hospital, Karachi, from January 2 to July 1, 2008. Two hundred pregnant women in their third trimester were included in the study. The diagnosis was based on history, clinical examination and relevant investigations. Patients with physiological dermatoses and dermatoses which flare up during pregnancy were excluded. A comprehensive pro-forma was used to evaluate different dermatoses. Skin biopsy for histopathology was also done where necessary. RESULTS: The age of the study population ranged from 17 to 36 years (mean = 27.3 +/- 4.86). Five (2.5%) patients had prurigo of pregnancy, 4 (2%) had dermatoses associated with Intrahepatic Cholestasis of Pregnancy (ICP), 3 (1.5%) patients had polymorphic eruption of pregnancy, and 1 (0.5%) patient had pruritic folliculitis of pregnancy. No case of pemphigoidgestationis was observed. CONCLUSION: In the study, 6.5% patients presented with specific dermatoses. Prurigo of pregnancy was the commonest condition. Polymorphic eruption of pregnancy was more common in primigravida, while dermatoses with intrahepatic cholestasis was seen more often in multigravida.

Role of intrinsic physicochemical parameters on multi-element distribution in surface sediment of the Devi River estuary, eastern India.

Multi-element composition including rare earth elements (REE) of surface sediment from the Devi river estuary, eastern coast of India, have been analysed in order to study the weathering characteristics and provenance of sediment along with their behaviour under different physicochemical conditions. These sediments with dominantly felsic provenance have undergone low to moderate chemical alterations. Bulk chemical composition is mainly represented by SiO2, Al2O3, Fe2O3 and K2O. Concentrations of Ba, Nb, Pb, Rb, Th and Zr are above their respective upper crustal abundances. High LREE/HREE ratio, negative Eu anomalies, and (La/Yb)n and (Tb/Yb)n values confirm that sediments are dominantly derived from the Eastern Ghat Group of rocks. Upper estuary sediments show negative Eu anomalies which is similar to that of the source. However, positive Eu anomaly is mostly observed in lower estuary. Contrasting Eu anomalies between upper- and lower-estuarine sediments are uncharacteristic of previously studied major global estuaries. Strong negative correlation between Mn and Eu suggests control of redox conditions over distribution of Eu. Concentration of REEs, Sc, Fe, Mo, V, Zn, Zr, Nb, U, Ti, Na and P increases up to 20 ppt salinity, and followed by declining trend towards mouth. This is mostly due to removal through flocculation of colloidal particles from water column during fresh- and saline-water interaction. This could be the first report about coagulation-based behaviour of Mo in estuarine environment. There is gradual decline in concentration of Cr, Co, Ni, Cu, Rb, Sr, Sb, Cs, Ba, Pb, Al, Mn, Mg, Ca and K with increase in salinity which is attributed to saline induced desorption of elements from sediments. The SiO2 content shows increasing trend towards mouth. Findings of this study highlight the importance of intrinsic physicochemical parameters, mainly salinity and redox condition, on governing geochemical behaviour of different elements including REE in mangrove dominated estuarine sediment.

Assessing the feasibility of evaluating and delivering a physical activity intervention for pre-school children: a pilot randomised controlled trial.

BACKGROUND: Few evidence-based physical activity interventions for pre-school children are available. This two-armed pilot cluster randomised controlled trial aimed to evaluate the feasibility of conducting a full-scale trial and of delivering an outdoor physical activity intervention for pre-school children. METHODS: School was the unit of randomisation, and follow-up occurred at 10 and 52 weeks. Trial feasibility was assessed by recruitment, retention and completion rates of primary (daily moderate-to-vigorous physical activity (MVPA)) and secondary (anthropometric, quality of life, self-efficacy) outcomes. Potential effectiveness was assessed for the primary outcome using a linear regression model comparing MVPA between trial arms adjusting for clustering by school. Feasibility of delivering the intervention was assessed by intervention fidelity and attendance. Semi-structured interviews with parents, intervention facilitators, and head teachers explored acceptability and capability to deliver the intervention as well as acceptability of the study design. RESULTS: Recruitment rates were 37 % of schools (n = 10 schools) and 48 % of pre-school children (n = 164 children). Retention of children to the trial at 52 weeks was 83.5 %. Thirty-nine percent of children had valid primary outcome accelerometer data at baseline and 52 weeks. Response rates for secondary outcome measures ranged from 52 to 88 % at 10 weeks and 59 to 80 % at 52 weeks. The mean difference in daily MVPA between trial arms at 52 weeks was 0.4, 95 % CI 16.3 to 17.0; p = 0.96. Fidelity of intervention implementation was 81 %. Intervention attendance was higher (82 %) during the summer initiation phase compared to autumn/spring initiation (50 %). Parents, facilitators and head teachers found the intervention acceptable and beneficial. CONCLUSIONS: Recruitment and retention rates suggest a trial in this outdoor setting with this population was feasible but is weather sensitive. However, strategies to increase accelerometer wear-time would need to be implemented for reliable primary outcome data to be obtained. There was high implementation fidelity by facilitators, and the intervention was seen as acceptable and deliverable. However, attendance was low and preliminary data showed no evidence of intervention effectiveness. A revised intervention, building on the successful elements of this pilot alongside adapting implementation strategies to improve attendance, should therefore be considered. TRIAL REGISTRATION: Trial registry name and number: Current Controlled Trials, ISRCTN54165860. Date of registration: 4 September 2012.

Sod1 deficiency reduces incubation time in mouse models of prion disease.

Prion infections, causing neurodegenerative conditions such as Creutzfeldt-Jakob disease and kuru in humans, scrapie in sheep and BSE in cattle are characterised by prolonged and variable incubation periods that are faithfully reproduced in mouse models. Incubation time is partly determined by genetic factors including polymorphisms in the prion protein gene. Quantitative trait loci studies in mice and human genome-wide association studies have confirmed that multiple genes are involved. Candidate gene approaches have also been used and identified App, Il1-r1 and Sod1 as affecting incubation times. In this study we looked for an association between App, Il1-r1 and Sod1 representative SNPs and prion disease incubation time in the Northport heterogeneous stock of mice inoculated with the Chandler/RML prion strain. No association was seen with App, however, significant associations were seen with Il1-r1 (P = 0.02) and Sod1 (P<0.0001) suggesting that polymorphisms at these loci contribute to the natural variation observed in incubation time. Furthermore, following challenge with Chandler/RML, ME7 and MRC2 prion strains, Sod1 deficient mice showed highly significant reductions in incubation time of 20, 13 and 24%, respectively. No differences were detected in Sod1 expression or activity. Our data confirm the protective role of endogenous Sod1 in prion disease.

"Pre-schoolers in the playground" an outdoor physical activity intervention for children aged 18 months to 4 years old: study protocol for a pilot cluster randomised controlled trial.

BACKGROUND: The pre-school years are considered critical for establishing healthy lifestyle behaviours such as physical activity. Levels of physical activity track through childhood into adulthood, thus establishing habitual physical activity early in life is vital. Time spent outdoors is associated with greater physical activity and playground interventions have been shown to increase physical activity in school aged children. There are few pre-school, playground-based interventions, and evaluations of these have found mixed results. A recent report published by the UK Chief Medical Officer (CMO) highlighted that new interventions to promote movement in the early years (0-5 years old) are needed. The aim of this study is to undertake a pilot cluster randomised controlled trial (RCT) of an outdoor playground-based physical activity intervention for parents and their children aged 18 months to 4 years old ("Pre-schoolers in the Playground"; PiP) and to assess the feasibility of conducting a full scale cluster RCT. The PiP intervention is grounded in behavioural theory (Social Cognitive Theory), and is in accordance with the CMO guidance for physical activity in the early years. It is informed by existing literature and data collected from focus groups with parents. METHODS/DESIGN: One hundred and fifty pre-school children affiliated to 10 primary schools will be recruited. Schools will be randomised to either the PiP intervention arm or the control arm (usual practice). Children in the intervention arm will be invited to attend three 30 minute outdoor play sessions per week for 30 weeks (3 school terms) at the school. Feasibility will be assessed by examining recruitment rates, attendance, attrition, acceptability of the trial and of the PiP intervention to parents, fidelity of intervention implementation, capability and capacity for schools to deliver the intervention. Health outcomes and the feasibility of outcome measurement tools will be assessed. These include physical activity via triaxial, accelerometry (Actigraph GT3X+), anthropometry (height, body mass, BMI, waist and upper arm circumference), health related quality of life for child (PedsQL) and parent (EQ5D), parent wellbeing (ComQol-A5), injuries and health service use. A health economic evaluation will also be undertaken. DISCUSSION: It is anticipated that results of this pilot trial will be published in spring 2015. TRIAL REGISTRATION: Current controlled trials: ISRCTN54165860.

Sex effects in mouse prion disease incubation time.

Prion disease incubation time in mice is determined by many factors including PrP expression level, Prnp alleles, genetic background, prion strain and route of inoculation. Sex differences have been described in age of onset for vCJD and in disease duration for both vCJD and sporadic CJD and have also been shown in experimental models. The sex effects reported for mouse incubation times are often contradictory and detail only one strain of mice or prions, resulting in broad generalisations and a confusing picture. To clarify the effect of sex on prion disease incubation time in mice we have compared male and female transmission data from twelve different inbred lines of mice inoculated with at least two prion strains, representing both mouse-adapted scrapie and BSE. Our data show that sex can have a highly significant difference on incubation time. However, this is limited to particular mouse and prion strain combinations. No sex differences were seen in endogenous PrP(C) levels nor in the neuropathological markers of prion disease: PrP(Sc) distribution, spongiosis, neuronal loss and gliosis. These data suggest that when comparing incubation times between experimental groups, such as testing the effects of modifier genes or therapeutics, single sex groups should be used.

The retinoic acid receptor beta (Rarb) region of Mmu14 is associated with prion disease incubation time in mouse.

In neurodegenerative conditions such as Alzheimer's and prion disease it has been shown that host genetic background can have a significant effect on susceptibility. Indeed, human genome-wide association studies (GWAS) have implicated several candidate genes. Understanding such genetic susceptibility is relevant to risks of developing variant CJD (vCJD) in populations exposed to bovine spongiform encephalopathy (BSE) and understanding mechanisms of neurodegeneration. In mice, aspects of prion disease susceptibility can be modelled by examining the incubation period following experimental inoculation. Quantitative trait linkage studies have already identified multiple candidate genes; however, it is also possible to take an individual candidate gene approach. Rarb and Stmn2 were selected as candidates based on the known association with vCJD. Because of the increasing overlap described between prion and Alzheimer's diseases we also chose Clu, Picalm and Cr1, which were identified as part of Alzheimer's disease GWAS. Clusterin (Clu) was considered to be of particular interest as it has already been implicated in prion disease. Approximately 1,000 heterogeneous stock (HS) mice were inoculated intra-cerebrally with Chandler/RML prions and incubation times were recorded. Candidate genes were evaluated by sequencing the whole transcript including exon-intron boundaries and potential promoters in the parental lines of the HS mice. Representative SNPs were genotyped in the HS mice. No SNPs were identified in Cr1 and no statistical association with incubation time was seen for Clu (P = 0.96) and Picalm (P = 0.91). Significant associations were seen for both Stmn2 (P = 0.04) and Rarb (P = 0.0005), however, this was only highly significant for Rarb. This data provides significant further support for a role for the Rarb region of Mmu14 and Stmn2 in prion disease.

Reconstruction of complex chest wall defects by using polypropylene mesh and a pedicled latissimus dorsi flap: a 6-year experience.

BACKGROUND: Reconstruction of full thickness defects of the chest wall is controversial and presents a complicated treatment scenario for thoracic and reconstructive plastic surgeons. It requires close cooperation between the cardiothoracic and reconstructive surgeons to achieve an optimal outcome and reduce the incidence of complications. OBJECTIVE: The purpose of this study is to evaluate our results in patients who underwent prosthetic bony reconstruction with polypropylene mesh and pedicle latissimus dorsi flap after chest wall resection. The principles of chest wall reconstruction include: wide excision of primary chest wall tumour with macroscopically healthy margins, wound excision and debridement of necrotic devitalised and irradiated tissues, control of infection and local wound care. STUDY DESIGN: This is a descriptive study. It includes 20 patients who underwent chest wall resection due to various causes and followed by reconstruction with polypropylene mesh along with pedicled latissimus dorsi flap. PLACE AND DURATION OF STUDY: The study was conducted at the Department of Plastic and Reconstructive Surgery, Federal Postgraduate Medical Institute, Sheikh Zayed Hospital Lahore, over a period of 6 years from August 1999 to August 2005. PATIENTS AND METHODS: This study included 20 patients who underwent chest wall reconstruction using polypropylene mesh and pedicled latissimus dorsi flap from August 1999 to August 2005. Patient demographic data including age, sex, pathological diagnosis, extent and type of resection, size of defect, and outcome were recorded. All patients were followed up in our outpatients department for 1 year. RESULTS: There was a total of 20 patients, 16 males and four females. The average age was 54 years (range 44-64 years). The indications for resection were primary chest wall tumours in 13 (65%) patients, local recurrence from breast tumours in one (5%) patient, post median sternotomy in three (15%) patients and radionecrosis in three (15%) patients. Ribs along with a part of sternum were resected in 14 (70%) patients, ribs along with clavicle in two (10%) patients and ribs only in four (20%) patients. The average area of chest wall defect after resection was 16.5 x 13 cm. In all patients, skeletal defect was reconstructed with polypropylene mesh. Soft tissue coverage was provided with a pedicled latissimus dorsi flap in all cases. Three patients with a chest wall tumour developed a recurrence within 6 months. Among these three, one patient died within 8 months of follow up due to myocardial infarction. CONCLUSION: Chest wall resection and reconstruction with synthetic polypropylene mesh and local muscle flaps can be performed as a safe, effective one-stage surgical procedure for a variety of major chest wall defects.

Versatility of the sural fasciocutaneous flap in the coverage of lower third leg and hind foot defects.

BACKGROUND: Reconstruction of soft tissue defects of the lower third of the leg, the heel and the hind foot remains a challenge. The distally based sural artery fasciocutaneous flap has been used effectively to resurface these defects. In many instances, it has obviated the need for free tissue transfer. OBJECTIVE: The objective of the study is to evaluate the efficacy of reverse sural artery fasciocutaneous flap for coverage of lower third leg, posterior heel, malleoli and hind foot. STUDY DESIGN: This is a descriptive study, which was conducted on 84 patients who presented with soft tissue defects in the area of lower third leg, heel, malleoli and hind foot. PLACE AND DURATION OF STUDY: The study was conducted at department of plastic and reconstructive surgery, Federal Postgraduate Medical Institute Shaikh Zayed Hospital Lahore, over a period of 7 years from February 1997 to February 2005. PATIENTS AND METHODS: Over a period of 7 years, a total of 84 patients with Soft tissue defect of lower third leg, heel, malleoli and hind foot were included. Preoperative data, the age and sex of each patient, cause and site of defect, dimension of flap, transposition of pedicle (through a tunnel or laid open and covered with a skin graft), postoperative results and complications were recorded. All patients were followed up in out patients department for 6 months. RESULTS: Out of 84 patients, 54 were males and 30 females. Their ages ranged from 8 to 55 years with a mean of 31 years. Road traffic accidents was the cause of the defects in 53 patients, wheel spoke injury in 12 patients, trophic ulcer in five patients, osteomyelitis in five patients, marjolin ulcer in seven patients and diabetic ulcer in two patients. The site of 84 defects comprised 52 distal tibia; 20 tendo-Achillis and posterior heel defects; seven-malleolar region; three-anterior ankle and two-foot amputation stumps. The dimension of flap ranged from 5 to 15 cm in length and 4 to 12 cm in width. Postoperatively 66 flaps survived completely while marginal necrosis was seen in six patients and infection in four patients. The complete flap necrosis occurred in eight patients. There was no considerable morbidity at donor site and all patients had satisfactory functional outcome. CONCLUSIONS: The distally based superficial sural artery flap is a versatile, reliable procedure, useful in reconstruction of lower third leg, heel, malleoli and hind foot defects. The surgical technique is safe, of short duration and provides alternative to microsurgical reconstruction.