Enhancement of heavy metal tolerance and accumulation efficiency by expressing Arabidopsis ATP sulfurylase gene in alfalfa.

Transgenic alfalfa (Medicago sativa L.) plants overexpressing the Arabidopsis ATP sulfurylase gene were generated using Agrobacterium-mediated genetic transformation to enhance their heavy metal accumulation efficiency. The ATP sulfurylase gene was cloned from Arabidopsis, following exposure to vanadium (V) and lead (Pb), and transferred into an Agrobacterium tumefaciens binary vector. This was co-cultivated with leaf explants of the alfalfa genotype Regen SY. Co-cultivated leaf explants were cultured on callus and somatic embryo induction medium, followed by regeneration medium for regenerating complete transgenic plants. The transgenic nature of the plants was confirmed using PCR and southern hybridization. The expression of Arabidopsis ATP sulfurylase gene in the transgenic plants was evaluated through RT-PCR. The selected transgenic lines showed increased tolerance to a mixture of five heavy metals and also demonstrated enhanced metal uptake ability under controlled conditions. The transgenic lines were fertile and did not exhibit any apparent morphological abnormality. The results of this study indicated an effective approach to improve the heavy metal accumulation ability of alfalfa plants which can then be used for the remediation of contaminated soil in arid regions.

Characterization of sida golden mottle virus isolated from Sida santaremensis Monteiro in Florida.

The genome of sida golden mottle virus (SiGMoV) (GU997691 and GU997692) isolated from Sida santaremensis Monteiro in Manatee County, Florida, was sequenced and characterized. SiGMoV was determined to be a bipartite virus belonging to the genus Begomovirus with a genome organization typical of the New World viruses in the genus. SiGMoV DNA-A had the highest identity scores (89%) and showed the closest evolutionary relationships to sida golden mosaic Buckup virus (SiGMBuV) (JX162591 and HQ008338). However, SiGMoV DNA-B had the highest identity scores (93%) and showed the closest evolutionary relationship to corchorus yellow spot virus (DQ875869), SiGMBuV (JX162592) and sida golden mosaic Florida virus (SiGMFlV) (HE806443). There was extensive recombination in the SiGMoV DNA-A and much less in DNA-B. Full-length clones of SiGMoV were infectious and were able to infect and cause symptoms in several plant species.

Microbial dynamics and properties of aerobic granules developed in a laboratory-scale sequencing batch reactor with an intermediate filamentous bulking stage.

Aerobic granules offer enhanced biological nutrient removal and are compact and dense structures resulting in efficient settling properties. Granule instability, however, is still a challenge as understanding of the drivers of instability is poorly understood. In this study, transient instability of aerobic granules, associated with filamentous outgrowth, was observed in laboratory-scale sequencing batch reactors (SBRs). The transient phase was followed by the formation of stable granules. Loosely bound, dispersed, and pinpoint seed flocs gradually turned into granular flocs within 60 days of SBR operation. In stage 1, the granular flocs were compact in structure and typically 0.2 mm in diameter, with excellent settling properties. Filaments appeared and dominated by stage 2, resulting in poor settleability. By stage 3, the SBRs were selected for larger granules and better settling structures, which included filaments that became enmeshed within the granule, eventually forming structures 2-5 mm in diameter. Corresponding changes in sludge volume index were observed that reflected changes in settleability. The protein-to-polysaccharide ratio in the extracted extracellular polymeric substance (EPS) from stage 1 and stage 3 granules was higher (2.8 and 5.7, respectively), as compared to stage 2 filamentous bulking (1.5). Confocal laser scanning microscopic (CLSM) imaging of the biomass samples, coupled with molecule-specific fluorescent staining, confirmed that protein was predominant in stage 1 and stage 3 granules. During stage 2 bulking, there was a decrease in live cells; dead cells predominated. Denaturing gradient gel electrophoresis (DGGE) fingerprint results indicated a shift in bacterial community composition during granulation, which was confirmed by 16S rRNA gene sequencing. In particular, Janthinobacterium (known denitrifier and producer of antimicrobial pigment) and Auxenochlorella protothecoides (mixotrophic green algae) were predominant during stage 2 bulking. The chitinolytic activity of Chitinophaga is likely antagonistic towards Auxenochlorella and may have contributed to stage 3 stable granule formation. Rhodanobacter, known to support complete denitrification, were predominant in stage 1 and stage 3 granules. The relative abundance of Rhodanobacter coincided with high protein concentrations in EPS, suggesting a role in microbial aggregation and granule formation.

Revision of type III and type IVB acetabular defects with Burch-Schneider anti-Protrusio cages.

Extensive bone loss in severe acetabular deficiencies can make repair with acetabular shells impossible. We retrospectively analyzed mid-term to long-term results of acetabular revision with Burch-Schneider reinforcement cages in 33 hips in patients with a mean age of 59.03years after a mean follow-up period of 6.19years. Under the American Academy of Orthopaedic Surgeons classification, 9 patients had type III acetabular bone defects and 24 had type IVB. Merle D'Aubigné scores improved by a mean of 9.3 points. The prosthesis failure rate was 15.1%, the series complication rate was 21.2%, and the mean survivorship was 11.57years. The Burch-Schneider cage is useful for reconstruction in massive acetabular deficiency, but complication rates are high and long-term mechanical failure is a concern.

Limits to sustained energy intake. XXII. Reproductive performance of two selected mouse lines with different thermal conductance.

Maximal sustained energy intake (SusEI) appears limited, but the factors imposing the limit are disputed. We studied reproductive performance in two lines of mice selected for high and low food intake (MH and ML, respectively), and known to have large differences in thermal conductance (29% higher in the MH line at 21°C). When these mice raised their natural litters, their metabolisable energy intake significantly increased over the first 13 days of lactation and then reached a plateau. At peak lactation, MH mice assimilated on average 45.3% more energy than ML mice (222.9±7.1 and 153.4±12.5 kJ day(-1), N=49 and 24, respectively). Moreover, MH mice exported on average 62.3 kJ day(-1) more energy as milk than ML mice (118.9±5.3 and 56.6±5.4 kJ day(-1), N=subset of 32 and 21, respectively). The elevated milk production of MH mice enabled them to wean litters (65.2±2.1 g) that were on average 50.2% heavier than litters produced by ML mothers (43.4±3.0 g), and pups that were on average 27.2% heavier (9.9±0.2 and 7.8±0.2 g, respectively). Lactating mice in both lines had significantly longer and heavier guts compared with non-reproductive mice. However, inconsistent with the 'central limit hypothesis', the ML mice had significantly longer and heavier intestines than MH mice. An experiment where the mice raised litters of the opposing line demonstrated that lactation performance was not limited by the growth capacity of offspring. Our findings are consistent with the idea that the SusEI at peak lactation is constrained by the capacity of the mothers to dissipate body heat.

Retraction: Intraosseous Calcaneal Lipoma Misdiagnosed as Plantar Fasciitis: An Orthopedic Case From Family Practice.

[This retracts the article DOI: 10.7759/cureus.21136.].

High Occurrence Among Calves and Close Phylogenetic Relationships With Human Viruses Warrants Close Surveillance of Rotaviruses in Kuwaiti Dairy Farms.

Rotavirus, one of the main pathogens causing morbidity and mortality in neonatal dairy calves worldwide, is responsible for 30-44% of cattle deaths. It is considered to be the most common etiologic agent of diarrhea in neonatal dairy calves and children, the dominant type being group A. Two hundred seventy animals from 27 farms from 2 regions of Kuwait were tested for the presence of Rotavirus serogroup A (RVA) using latex agglutination test (LAT) and reverse transcription-polymerase chain (RT-PCR) testing. RVA non-structural proteins NSP1-2, NSP4-5 and capsid protein genes VP1-7 were characterized by next generation sequencing. LAT was positive in 15.56% of the animals, and RT-PCR in 28.89%. Using RT-PCR as a reference method, LAT was 100% specific but only 83.33% sensitive. ANOVA analysis showed correlation only with the location of the farms but no significant correlation with the age and sex of the animals. Although there was a tendency of clustering of RVA positive animals, it did not reach statistical significance (p = 0.035 for LAT). The phylogenetic analysis showed that Kuwaiti isolates of group A rotavirus clustered with human rotaviruses. Taken together, it seems that rotavirus was present in most of the dairy farms in Kuwait. The high occurrence of the virus in calves in Kuwaiti dairy farms and the close phylogenetic affinity with human isolates warrants urgent action to minimize and control its spread between calves in farms.

Survival From Ninety-Five Percent Total Body Surface Area Burn: A Case Report and Literature Review.

Burns can be devastating and result in unwanted consequences with prolonged length of hospital stay. The mortality rate increases as the total body surface area increases, so proper management of patients with extensive degrees of burns is crucial for their survival. We present the hospital course, management, and survival of a patient after he sustained a 95% total body surface area, second-degree burn from a gas flame. Furthermore, we present from the literature different cases of patients with large total body surface area burns and survived after being managed in burns specialty centers. Although large total body surface area burns can result in significant morbidity and mortality, early management and intervention by an expert surgical team can result in positive outcomes.

Intraosseous Calcaneal Lipoma Misdiagnosed as Plantar Fasciitis: An Orthopedic Case From Family Practice.

Foot pain is a prevalent musculoskeletal complaint with plantar fasciitis being one of the most common underlying etiologies. Diagnostic imaging studies are rarely performed for patients with plantar fasciitis. We report the case of a 34-year-old woman with a six-month history of left heel pain that was worsened by prolonged standing and walking. She was diagnosed as having plantar fasciitis. Considering the lack of clinical improvement on conservative measures, the patient underwent a plain radiograph, which demonstrated a geographic radiolucent lesion in the calcaneus with well-defined non-sclerotic margins. Subsequently, a computed tomography scan re-demonstrated the lesion as having a fat-attenuation appearance that was classic of lipoma. The patient underwent surgical excision of the tumor and the histopathological examination of the specimen revealed spindle cell lipoma. The patient had complete resolution of the pain following the surgery. Intraosseous lipoma is a very rare benign primary bone tumor. The case demonstrated that the intraosseous lipoma may have similar clinical features to plantar fasciitis. Hence, physicians should keep a high index of suspicion for the alternate diagnoses of plantar fasciitis in patients who do not exhibit any clinical improvement despite a prolonged period of conservative management.

Comparative Agronomic Performance and Reaction to Fusarium wilt of Lens culinaris × L. orientalis and L. culinaris × L. ervoides derivatives.

The development of transgressive phenotype in the segregating populations has been speculated to contribute to niche divergence of hybrid lineages, which occurs most frequently at larger genetic distances. Wild Lens species are considered to be more resistant against major biotic and abiotic stresses than that of the cultivated species. In the present study, we assessed the comparative agronomic performance of lentil (Lens culinaris subsp. culinaris) inter-sub-specific (L. culinaris subsp. orientalis) and interspecific (L. ervoides) derivatives, also discussed its probable basis of occurrence. The F3, F4, and F5 inter sub-specific and interspecific populations of ILL8006 × ILWL62 and ILL10829 × ILWL30, respectively revealed a substantial range of variation for majority of agro-morphological traits as reflected by the range, mean and coefficient of variation. A high level of fruitful heterosis was also observed in F3 and F4 progeny for important traits of interest. Phenotypic coefficient of variation (PCV) was higher in magnitude than genotypic coefficient of variation (GCV) in all generations for several quantitative characters. The results showed high heritability estimates for majority of traits in conjunction with low to high genetic advance in F3 and F4 generations. Further, F5 progeny of ILL10829 × ILWL30, manifested resistant disease reaction for fifteen recombinant inbred lines (RILs) against (Fusarium oxysporum f. sp. lentis (Vasd. Srin.) Gord.). The multilocation agronomic evaluation of both crosses showed better results for earliness, desirable seed yield and Fusarium wilt resistance under two agro-ecological regions of north-western India. These better performing recombinants of ILL8006 × ILWL62 and ILL10829 × ILWL30 can be advanced for further genetic improvement and developing high yielding disease resistant cultivars of lentil.

Oxidative costs of reproduction in mouse strains selected for different levels of food intake and which differ in reproductive performance.

Oxidative damage caused by reactive oxygen species has been hypothesised to underpin the trade-off between reproduction and somatic maintenance, i.e., the life-history-oxidative stress theory. Previous tests of this hypothesis have proved equivocal, and it has been suggested that the variation in responses may be related to the tissues measured. Here, we measured oxidative damage (protein carbonyls, 8-OHdG) and antioxidant protection (enzymatic antioxidant activity and serum antioxidant capacity) in multiple tissues of reproductive (R) and non-reproductive (N) mice from two mouse strains selectively bred for high (H) or low (L) food intake, which differ in their reproductive performance, i.e., H mice have increased milk energy output (MEO) and wean larger pups. Levels of oxidative damage were unchanged (liver) or reduced (brain and serum) in R versus N mice, and no differences in multiple measures of oxidative protection were found between H and L mice in liver (except for Glutathione Peroxidase), brain or mammary glands. Also, there were no associations between an individual's energetic investment (e.g., MEO) and most of the oxidative stress measures detected in various tissues. These data are inconsistent with the oxidative stress theory, but were more supportive of, but not completely consistent, with the 'oxidative shielding' hypothesis.

The epidemiology of syphilis in pregnancy.

This study evaluated the magnitude, risk factors and outcomes of syphilis in pregnancy in a large cohort of women in four countries participating in the World Health Organization (WHO) antenatal care trial. All women attending the first prenatal care at each selected clinic were enrolled. Screening at the first antenatal visit was routinely performed with either rapid plasma reagin or Venereal Disease Research Laboratory and confirmed by fluorescent treponemal antibody absorption. All women also had the same syphilis tests after delivery. The initial prevalence, the incidence during pregnancy and the overall prevalence of syphilis at delivery were 0.9%, 0.4% and 1.3% respectively. Risk factors for syphilis during pregnancy were younger age for the incidence and older age and a history of stillbirth for the prevalence. Women with syphilis during pregnancy had significantly more adverse outcomes. We support the recommendation that in addition to the initial testing, a second routine test for syphilis ought to be established early in the third trimester even in low prevalence areas.

Risk factors for spontaneous preterm birth in a Saudi population.

Preterm birth (< 37 weeks completed gestation), the primary predictor of infant morbidity and mortality, can result from diverse biologic and sociodemographic variables. A case-control study was undertaken to determine risk factors that were significantly associated with preterm birth in our population. Pertinent data were collected by structured interviews with eligible subjects and by medical record abstraction. The study population consisted of 118 mothers of singleton, preterm, appropriate for gestational age infants (cases) and 118 mothers of singleton, term, appropriate for gestational age infants (controls). A multiple logistic regression analysis indicated that significant risk factors for preterm birth were first or second trimester vaginal bleeding during current pregnancy, a previous preterm birth, inadequate prenatal care, consanguinity, maternal body mass index of < 23, and short inter-pregnancy interval. Awareness of such risk factors is essential in planning public education programs and in considering appropriate perinatal care options for women at potentially higher risk for preterm delivery.

The routine use of cefazolin in cesarean section.

OBJECTIVE: To determine the effectiveness and safety of the routine use of antibiotic prophylaxis in women undergoing cesarean section. METHOD: Four hundred and forty-one women undergoing cesarean sections were randomly assigned either to a single dose of 1 g intravenous cefazolin or placebo after clamping of the umbilical cord. The primary outcome was the development of post-operative febrile morbidity and the secondary outcomes were infection-related complications. RESULT: There were 211 emergency and 230 elective cesarean sections. In the emergency cesarean sections, 34 (30.6%) women developed post-operative febrile morbidity in the placebo group compared to 11 (11%) women in the cefazolin group. This was a statistically significant difference (P = 0.001). Similarly, there were statistically significant differences between the two groups in the development of wound infection (P<0.001), use of therapeutic antibiotics (P = 0.001), and post-operative days in hospital (P = 0.003). No statistically significant differences were detected in the development of other infection-related complications. In the elective cesarean sections, no statistically significant differences were found in post-operative febrile morbidity and infection-related complications. There were no serious side effects related to the use of cefazolin. CONCLUSION: The routine use of a single dose of cefazolin is safe and effective in emergency but not elective cesarean section.

Post partum thyroiditis in Saudi women.

Two hundred and seventy seven Saudi women were prospectively evaluated post-partum to determine the frequency of post-partum thyroiditis. Four to six weeks after delivery, 12 (4.3%) had positive antimicrosomal antibodies and 8 (2.9%) had positive antithyroglobulin antibodies. At 6-8 weeks post-partum, out of 82 subjects followed up, 4 (4.9%) had positive antimicrosoMal antibodies and 1 (1.2%) had positive antithyroglobulin antibodies. At 8-12 weeks post-partum, out of 70 subjects, antimicrosomal antibodies were detected in 5 (3.5%) subjects and antithyroglobulin in 2 (1.4%). Their routine thyroid function tests (T3, T4, and TSH) remained within the normal range. There was no clinical evidence of hyper- or hypothyroidism in any of the patients during the follow up. Post-partum thyroiditis, appears to be relatively uncommon in Saudi Arabia. It does not produce any clinical illness or cause significant subclinical thyroid dysfunction.

Low birthweight in Riyadh, Saudi Arabia: incidence and risk factors.

In a multicentre prospective study, we have determined the incidence of low birthweight (LBW) and the main predisposing risk factors. Among 4651 consecutive births over a 5-month period in five hospitals in Riyadh, the overall incidence of LBW was 8.4%. When stillbirths were excluded the incidence of LBW was 7.4% of all live births. Statistical analysis was performed among 638 births (319 LBW infants, i.e. less than or equal to 2499 g and 319 babies weighing 2500 g or more). Of the 28 antenatal risk variables analysed, 13 were found to be significant when studied separately. Of these 13 variables, six were found to be significant predictors of LBW, using stepwise multiple logistic regression. These six variables together correctly predicted 72% and 88% of the LBW babies or normal birthweight babies, respectively. The risk factors thus identified were (i) short gestation, (ii) multiple gestation, (iii) low maternal body mass index, (iv) nulliparity, (v) availability of housework help, and (vi) absence of consanguinity. Measures for reducing these factors are also discussed.

Epidermal clitoral inclusion cyst after type I female genital mutilation.

OBJECTIVE: To document the occurrence of long-term sequelae after type I female genital mutilation (FGM) and describe the surgical treatment of epidermal clitoral inclusion cyst. STUDY DESIGN: Twenty-one women presented with epidermal clitoral inclusion cyst after type I FGM at the Department of Obstetrics and Gynecology at King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia. The duration (mean +/- SD, range) of symptoms was 10.3 +/- 5.4, 2 to 20 years. They were treated by excision of the cyst with particular attention to preserve the remaining part of the clitoris. The technique involves making a vertical incision in the skin, dissecting and excising the cyst, removing the excessive skin, and reapproximating the skin edges. RESULTS: The procedure was done on all patients without intraoperative complications. All except one were discharged home on the second postoperative day. Follow-up showed no recurrence of symptoms. CONCLUSION: Long-term sequelae can occur after type I FGM. The surgical treatment of clitoral inclusion cyst is simple and effective.

The practice of antenatal care: comparing four study sites in different parts of the world participating in the WHO Antenatal Care Randomised Controlled Trial.

In the preparation of a randomised controlled trial to evaluate a new programme of antenatal care (ANC) in different parts of the world, we conducted a baseline survey of the ANC procedures in all 53 clinics participating in the trial. There were two components of this survey: (1) description of clinic characteristics and services offered: the staff of each clinic was interviewed and direct observation was made by field supervisors, and (2) the actual use of services by pregnant women attending these clinics: we reviewed a random sample of 2913 clinical histories. The clinical units surveyed were offering most of the activities, screening, laboratory tests and interventions recommended as effective according to the Cochrane Pregnancy and Childbirth Database (PCD), although some of these were not available in some sites. On the other hand, some tests and interventions that are considered not effective according to these criteria are reportedly offered. There was a difference across sites in the availability and offer to low-risk women of vaginal examination, evaluation of pelvic size, dental examination, external version for breech presentation and formal risk score classification, and a notable difference in the type of principal provider of ANC. There was a large variation in the actual use of screening and laboratory tests and interventions that should be offered to all women according to Cochrane PCD criteria: some of these are simply not available in a site; others are available, but only a fraction of women attending the clinics are receiving them. The participating sites all purport to follow the traditional 'Western' schedule for ANC, but in three sites we found that a high percentage of women initiate their ANC after the first trimester, and therefore do not have either the recommended minimum number of visits during pregnancy or the minimum first trimester evaluation. It is concluded that the variability and heterogeneity of ANC services provided in the four study sites are disturbing to the profession and cast doubts on the rationale of routine ANC.

PIP: This paper reports the results of an antenatal care (ANC) baseline survey in four study sites in different parts of the world participating in the WHO ANC randomized controlled trial. The health care units surveyed are providing most of the activities, screening, laboratory tests and interventions recommended as effective based on the Cochrane Pregnancy and Childbirth Database (PCD), although some of these are not available in some clinics. Some tests and interventions that are considered ineffective according to the PCD criteria are reportedly offered. A variation is observed across sites in the availability and offer to low-risk women of vaginal exam, pelvic size assessment, dental exam, external version for breech presentation, and formal risk score classification. Moreover, there is a significant difference in the type of principal ANC provider and a great variation in the actual performance of screening and laboratory tests and interventions that should be provided to all women according to these criteria. This is partly due to the unavailability of these services in the site, or only a fraction of women attending the clinics are receiving them. The participating clinics all declared to follow the traditional Western schedule for ANC; however, three clinics are found to have a high percentage of women who begin their ANC after the first trimester of pregnancy. It is generalized that the variability and heterogeneity of ANC services offered in the four sites are annoying to the profession and cast doubts on the rationale behind routine ANC.

Determinants of term intrauterine growth retardation: the Saudi experience.

In a clinical study from an unselected Saudi obstetric population, the incidence of and risk factors for intrauterine growth retardation among live births were investigated. From a total study group of 4578 consecutive live births, 76 (1.7%) infants were found to be growth retarded. These infants were then compared with a randomly selected control group of 76 term newborns with appropriate birthweight for their gestational ages. Delivery at term of a growth-retarded infant was significantly associated with maternal age under 20 years, maternal body mass index less than 23, first degree consanguinity, poor housing, primiparity, and inadequate prenatal care in univariate analysis. When considered jointly in multivariate logistic regression analysis, the significant determinants were reduced to primiparity, first degree consanguinity, and poor housing. These risk factors correctly predicted 63% and 71% of the intrauterine growth-retarded infants or normal birthweight infants, respectively.

PIP: In Saudi Arabia, physicians compared data on 76 term intrauterine growth retardation (IUGR) infants with data on 76 randomly selected term newborns of appropriate birth weight to study the determinants of term IUGR. The term IUGR rate was 1.7% (76/4578 consecutive live births). The univariate analysis found significant risk factors to be a maternal age of less than 20 years (odds ratio [OR] = 4.89), a maternal body mass index of less than 23 (OR = 2.86), first degree consanguinity (OR = 3.1), living in a mud house (OR = 5.10), primiparity (OR = 3), and inadequate prenatal care (OR = 2.86). A stepwise multiple logistic regression model revealed that the significant risk factors of term IUGR included primiparity (OR = 3.3), first degree consanguinity (OR = 3.4), and living in a mud house (OR = 7.5). These three risk factors correctly predicted 63% and 71% of the IUGR and normal-birth-weight infants, respectively. These findings suggest that health care providers may be able to identify pregnancies at risk of IUGR and can thus prevent and manage IUGR births.