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1.  Thyroid dysfunction among type 2 diabetic female Egyptian subjects 
High prevalence of thyroid disorders is more common in type 1 diabetes compared to type 2 diabetes, due to associated autoimmunity. Hypothyroidism is the most common disorder. The objective was to assess the prevalence of thyroid dysfunction among type 2 diabetic Egyptian females and to find the correlation between metabolic syndrome components and autoimmune thyroid dysfunction.
Materials and methods
The study included 62 type 2 diabetic Egyptian females and 27 sex- and age-matched controls. All patients in the study were subjected to anthropometric measures, including HbA1c, lipid profile, serum uric acid, thyroid-stimulating hormone (TSH), free triiodothyronine, free thyroxine, anti-thyroid peroxidase (TPO), antithyroglobulin (anti-Tg), and thyroid ultrasound.
Hypothyroidism was found in 45.2% of patients (5.49±3.37 μIU/mL) versus 11.1% of controls (1.79±1.21 μIU/mL) (P<0.001). Anti-TPO was found in 75.8% (347.15±244.87 IU/mL) of patients versus 7.4% (32.89±33.26 IU/mL) of controls (P<0.001). Anti-Tg was found in 61.3% (508.03±369.16 IU/mL) of patients versus 0 (51.26±35.53 IU/mL) controls (P<0.001). A significant positive correlation was found between TSH and antithyroid antibodies (anti-Tg, anti-TPO; P=0.002 and P=0.043, respectively) and between TSH and thyroid-gland volume (P=0.002) in diabetic patients. No correlation was found between any components of metabolic syndrome and thyroid antibodies in diabetic patients.
Autoimmune thyroid disease is more common in Egyptian women with type 2 diabetes than nondiabetic women, and thus points to a role of autoimmunity in the pathogenesis of type 2 diabetes.
PMCID: PMC5123662  PMID: 27920545
autoimmune thyroid dysfunction; TSH; anti-TPO; anti-Tg; T2 diabetes; metabolic syndrome
2.  Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples 
Journal of Parasitology Research  2016;2016:7376207.
Ascaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise. We have developed a loop mediated isothermal amplification (LAMP) for diagnosis of ascariasis in fecal samples, based on the first internal transcribed (ITS-1) spacer region of the ribosomal DNA. We used Primer Explorer V4 software to design primers. Ascaris adult and ova were obtained from naturally infected school children, whose parents/guardians gave consent for their participation in the study. Genomic DNA was extracted using alkaline lysis method and amplified by LAMP at 63°C for 45 minutes. LAMP products were visualized by naked eyes after adding SYBR Green dye and also on agarose gel. LAMP successfully and reliably detected Ascaris DNA from a single egg and in fecal samples. The assay specifically detected Ascaris DNA without amplifying DNA from ova of other parasites which commonly coexist with A. lumbricoides in feces. The developed LAMP assay has great potential for use in ascariasis diagnosis at the point of care and in low infection intensity situation that characterize control and elimination campaigns.
PMCID: PMC5108867  PMID: 27882242
3.  Prevalence of irritable bowel syndrome among Qassim school teachers, and its impact on their performance and life duties 
Saudi Medical Journal  2016;37(7):817.
To determine the prevalence of irritable bowel syndrome (IBS) among school teachers in Qassim region, Kingdom of Saudi Arabia (KSA) and it’s impact on their performance -as teachers- and on life duties.
A cross-sectional study was held in Qassim region, KSA between 2012 and 2013. The study was conducted among 300 school teachers selected by multistage stratified random sample method in 2013. The sample size was calculated according to the formula: n = (Z2 * (p) * (q))/c2. A confidential, anonymous, and a self-administered questionnaire was used to collect personal and sociodemographic data, teacher performance oriented questions were included. Rome III Criteria were used. Teachers with red flag symptoms were excluded.
The prevalence of IBS symptoms among teachers in Qassim region of KSA is 40.7% with no significant gender predilection. The IBS symptoms significantly affect the work performance of the teachers (61.6%) with high absenteeism rate (46.6%), especially among 51-60 years and 21-30 years, age groups (p<0.05 in chi-square test). Significant proportion (45.3%) of IBS criteria +ve teachers find IBS symptoms to be hindrance in their life duties.
Prevalence of IBS in Qassim school teachers is becoming an important issue with 40.7% of teachers reporting symptoms suggestive of IBS in a random sample. Majority of teachers are having frequent performance problems - as teachers - and in usual life duties associated with their IBS Symptoms. Significantly, IBS criteria +ve teachers are more likely to have discipline problems compared with healthy teachers. We recommend a health care program to tackle the problem among the teachers, to improve their performance and reduce working hours loss. Also, follow up studies need to be conducted to assess the health care of IBS patients in local public health care centers.
PMCID: PMC5018652  PMID: 27381548
4.  Inflammatory Breast Cancer: A Distinct Clinicopathological Entity Transcending Histological Distinction 
PLoS ONE  2016;11(1):e0145534.
Although well recognized in breast oncology literature, histologic subtypes have not been previously described in inflammatory breast cancer (IBC). The purpose of this study was to describe lobular subtype in IBC and assess the impact of histology on patient outcomes.
We performed a retrospective analysis of 659 IBC patients at MD Anderson Cancer Center between January 1984 and December 2009. Patients with Invasive Lobular, Mixed Invasive Ductal and Lobular, or Invasive Ducal Carcinomas (ILC, MIC, IDC, respectively) comprise the subject of this report. Patient characteristics and survival estimates were compared by using chi-square test and Kaplan-Meier method with log-rank statistic. Cox proportional hazards models were fit to determine association of histology with outcomes after adjustment for other characteristics.
A total of 30, 37, and 592 patients were seen to have invasive lobular, mixed, or ductal histology, respectively. Grade 3 tumors were more common in the ductal group (78%) than in the lobular (60%) or mixed (61%) group (P = 0.01). The 3-year overall survival rates were 68%, 64%, and 62% in the lobular, mixed, and ductal groups, respectively (P = 0.68). After adjustment, histology did not have a significant effect on death in the lobular group (HR = 0.70, 95% confidence interval [CI]: 0.26–1.94; P = 0.50) or mixed group (HR = 0.53, 95% CI: 0.25–1.13; P = 0.10) compared with the ductal group.
In this cohort of IBC patients, lobular histology was seen in 4.5% cases. Histology does not appear to have a significant effect on survival outcomes in IBC patients, unlike in patients with non-inflammatory breast cancer (n-IBC), indicating the distinct biological behavior of the IBC phenotype.
PMCID: PMC4709074  PMID: 26752563
5.  Development and evaluation of a Loop Mediated Isothermal Amplification (LAMP) technique for the detection of hookworm (Necator americanus) infection in fecal samples 
Parasites & Vectors  2015;8:574.
Hookworm infection is a major concern in sub-Saharan Africa, particularly in children and pregnant women. Necator americanus and Ancylostoma duodenale are responsible for this condition. Hookworm disease is one of the Neglected tropical diseases (NTDs) that are targeted for elimination through global mass chemotherapy. To support this there is a need for reliable diagnostic tools. The conventional diagnostic test, Kato-Katz that is based on microscopic detection of parasite ova in faecal samples, is not effective due to its low sensitivity that is brought about mainly by non-random distribution of eggs in stool and day to day variation in egg output. It is tedious, cumbersome to perform and requires experience for correct diagnosis. LAMP-based tests are simple, relatively cheap, offer greater sensitivity, specificity than existing tests, have high throughput capability, and are ideal for use at the point of care.
We have developed a LAMP diagnostic test for detection of hookworm infection in faecal samples. LAMP relies on auto cycling strand displacement DNA synthesis performed at isothermal temperature by Bst polymerase and a set of 4 specific primers. The primers used in the LAMP assay were based on the second Internal Transcribed Spacer (ITS-2) region and designed using Primer Explorer version 4 Software. The ITS-2 region of the ribosomal gene (rDNA) was identified as a suitable target due to its low mutation rates and substantial differences between species. DNA was extracted directly from human faecal samples, followed by LAMP amplification at isothermal temperature of 63 °C for 1 h. Amplicons were visualized using gel electrophoresis and SYBR green dye. Both specificity and sensitivity of the assay were determined.
The LAMP based technique developed was able to detect N. americanus DNA in faecal samples. The assay showed 100 % specificity and no cross-reaction was observed with other helminth parasites (S. mansoni, A. lumbricoides or T. trichiura). The developed LAMP assay was 97 % sensitive and DNA at concentrations as low as 0.4 fg were amplified.
The LAMP assay developed is an appropriate diagnostic method for the detection of N. americanus DNA in human stool samples because of its simplicity, low cost, sensitivity, and specificity. It holds great promise as a useful diagnostic tool for use in disease control where infection intensities have been significantly reduced.
PMCID: PMC4636844  PMID: 26546069
Loop mediated isothermal amplification; ITS-2; Necator americanus; Kato-Katz
6.  Loin to groin pain: The importance of a differential diagnosis 
•We present a rare case of exertional paravertebral lumbar compartment syndrome that was initially misdiagnosed as ureteric colic.•Exertional lumbar paravertebral compartment syndrome typically present with intractable lumbar back pain 6–12 h following lumbar specific exercise.•There is currently no consensus on management with both conservative and surgical techniques used and case reports the only standard for comparison.•Long term back pain is more likely in patients managed conservatively.
Ureteric colic frequently presents as loin to groin pain and accounts for a significant proportion of emergency urological admissions. However, a number of differential diagnoses should be considered in a systematic approach when assessing patients.
Presentation of case
We report a case of a 30 year old man admitted with severe unilateral loin to groin pain following lumbar specific weightlifting exercises. After a significant delay due to initial mis-diagnosis he was diagnosed with acute paravertebral lumbar compartment syndrome (PVCS) and managed conservatively.
Exertional PVCS is a rare and potentially life threatening condition arising following lumbar specific exercise that has only been recorded a handful of times previously. Patients typically present with intractable lumbar pain and rhabdomyolysis 6–12 h following exercise. Due to initial diagnostic delay our case was managed conservatively with fluid resuscitation and monitoring of renal function.
Assessment of patients with loin pain requires a systematic approach. PVCS is a rare cause of lumbar back and loin pain but one that should be considered, particularly in active young males. Early diagnosis is key to prevent the potential sequalae of untreated rhabdomyolysis. There is currently no consensus on management option for PVCS with only a few cases being reported in the literature. We describe successful management with supportive non operative treatment.
PMCID: PMC4643462  PMID: 26453939
Paravertebral; Compartment; Syndrome; Rhabdomyolysis; Colic
7.  Assessment of metal artefact reduction around dental titanium implants in cone beam CT 
Dentomaxillofacial Radiology  2014;43(7):20140019.
The aim of this study was to investigate if the metal artefact reduction (MAR) tool used in the software of the ORTHOPANTOMOGRAPH® OP300 (Instrumentarium Dental, Tuusula, Finland) can improve the gray value levels in post-operative implant scans.
20 potential implant sites were selected from 5 edentulous human dry mandibles. Each mandible was scanned by a CBCT scanner, and images were produced under three different conditions: implant sites drilled but no implants inserted, implants inserted without application of MAR and implants inserted with application of MAR. Using Geomagic® Studio 2012 (Geomagic, Morrisville, NC) and 3Diagnosys® v. 5.3.1 (3Diemme® SRL, Cantù, Italy) software, three scans of each mandible were superimposed. The mean gray value of identical regions of bone around the implants was derived for each condition. The differences between gray value measurements at implant sites derived from different conditions were assessed.
A significant difference was found between mean gray values from the scans with no implants inserted and with implants inserted (with and without MAR) (p = 0.012). No significant difference was revealed for gray values measured from scans with and without MAR (p = 0.975).
The MAR tool in the software of the ORTHOPANTOMOGRAPH OP300 CBCT scanner does not significantly correct the voxel gray values affected by the metal artefact in the vicinity of an implant in human dry mandibles.
PMCID: PMC4170846  PMID: 25135316
cone beam computed tomography; artefacts; dental implants; titanium
8.  Non-trauma surgical emergencies in adults: Spectrum, challenges and outcome of care 
Annals of Medicine and Surgery  2015;4(4):325-330.
Significant deaths of between 21% and 38% occur from non-trauma surgical conditions in the accident and emergency room. Access to emergency surgical care is limited in many developing countries including Nigeria. We aimed to study the spectrum of non-trauma surgical emergencies, identify challenges in management and evaluate outcomes.
A one year prospective cohort study of all non-trauma emergencies in adults seen at the surgical emergency room of LASUTH from 1st October, 2011 to 30th September, 2012 was conducted. Data was analyzed using SPSS version 15.0.
Of a total of 7536 patients seen, there were 7122 adults. Those with non-trauma conditions were 2065 representing 29% of adult emergencies. Age ranged between 15 and 97 years and male to female ratio was 1.7:1. Acute abdomen (30%), urological problems (18%) and malignancies (10%) were the most common. Among 985 patients requiring admission only 464 (47%) were admitted while the remaining 53% were referred to other centers. Emergency surgical intervention was carried out in 222 patients representing 48% of admitted patients. There were 12 (24%) non-trauma deaths in the emergency room. They were due to acute abdomen and malignancies in half of the cases.
Facilities for patients needing emergency care were inadequate with more than half of those requiring admission referred. Attention should be paid to the provision of emergency surgical services to the teeming number of patients seen on yearly basis in the Teaching Hospital.
•Non-trauma conditions constitute a third of surgical emergencies and a quarter of deaths in the emergency room.•Acute abdomen, urological conditions and malignancies are the leading causes of non-trauma emergencies.•Available facilities could provide care for only half of the non-trauma patients requiring emergency room admission.
PMCID: PMC4600937  PMID: 26566434
Facility; Non-trauma; Emergencies; Outcome; Spectrum
9.  Open-label randomized clinical trial of standard neoadjuvant chemotherapy with paclitaxel followed by FEC versus the combination of paclitaxel and everolimus followed by FEC in women with triple receptor-negative breast cancer† 
Annals of Oncology  2014;25(6):1122-1127.
Phase II randomized study to determine pharmacodynamics (PD) changes and response of adding everolimus to standard neoadjuvant chemotherapy in triple-negative breast cancer (TNBC).
Everolimus synergistically enhances taxane-induced cytotoxicity in breast cancer cells in vitro and in vivo in addition to demonstrating a direct antiproliferative activity. We aim to determine pharmacodynamics changes and response of adding everolimus to standard neoadjuvant chemotherapy in triple-negative breast cancer (TNBC).
Patients and methods
Phase II study in patients with primary TNBC randomized to T-FEC (paclitaxel 80 mg/m2 i.v. weekly for 12 weeks, followed by 5-fluorouracil 500 mg/m2, epirubicin 100 mg/m2, and cyclophosphamide 500 mg/m2 every 3 weeks for four cycles) versus TR-FEC (paclitaxel 80 mg/m2 i.v. and everolimus 30 mg PO weekly for 12 weeks, followed by FEC). Tumor samples were collected to assess molecular changes in the PI3K/AKT/mTOR pathway, at baseline, 48 h, 12 weeks, and at surgery by reverse phase protein arrays (RPPA). Clinical end points included 12-week clinical response rate (12-week RR), pathological complete response (pCR), and toxicity.
Sixty-two patients were registered, and 50 were randomized, 27 received T-FEC, and 23 received TR-FEC. Median age was 48 (range 31–75). There was downregulation of the mTOR pathway at 48 h in the TR-FEC arm. Twelve-week RR by ultrasound were 29.6% versus 47.8%, (P = 0.075), and pCR were 25.9% versus 30.4% (P = 0.76) for T-FEC and TR-FEC, respectively. mTOR downregulation at 48 h did not correlate with 12-week RR in the TR-FEC group (P = 0.58). Main NCI grade 3/4 toxicities included anemia, neutropenia, rash/desquamation, and vomiting in both arms. There was one case of grade 3 pneumonitis in the TR-FEC arm. No grade 3/4 stomatitis occurred.
The addition of everolimus to paclitaxel was well tolerated. Everolimus downregulated mTOR signaling but downregulation of mTOR at 48 h did not correlate with 12-week RR in the TR-FEC group.
Clinical trial number
PMCID: PMC4037860  PMID: 24669015
triple negative breast cancer; neoadjuvant systemic therapy; PI3K pathway inhibition
10.  The Structure of Clinical Consultation: A Case of Non-Native Speakers of English as Participants 
Global Journal of Health Science  2014;7(1):249-260.
In many parts of the world, patients may find it difficult to visit doctors who share the same language and culture due to the intermingling of people and international recruitment of doctors among many other reasons. In these multilingual multicultural settings (MMSs), doctor-patient interactions face new communication challenges. This study aims to identify the structure of clinical consultation and its phases in an MMS where both doctors and patients are non-native speakers (NNSs) of English.
This study takes on a discourse analytic approach to examine the structure of clinical consultation as an activity type. 25 clinical consultation sessions between non-native speakers of English in a public healthcare centre in Malaysia were audio-recorded.
Findings and Discussion:
The results show that there are some deviations from the mainstream structure of clinical consultations although, in general, the pattern is compatible with previous studies. Deviations are particularly marked in the opening and closing phases of consultation.
In almost all interactions, there is a straightforward manner of beginning medical consultations. The absence of greetings may have naturally reduced the length of talk. Hence, by directly entering medical talks, the doctors voice their concern on the curing aspects of the consultation rather than its caring facets. The preference of curing priority to caring is more goal-oriented and in alignment with the consultation as an activity type.
PMCID: PMC4796523  PMID: 25560336
clinical consultation; doctor-patient interaction; multilingualism; multiculturalism; non-native speaker; misunderstanding
11.  The effect of scan parameters on cone beam CT trabecular bone microstructural measurements of the human mandible 
Dentomaxillofacial Radiology  2013;42(10):20130206.
The objective of this study was to investigate the effect of different cone beam CT scan parameters on trabecular bone microstructure measurements. A human mandibular cadaver was scanned using a cone beam CT (3D Accuitomo 170; J.Morita, Kyota, Japan). 20 cone beam CT images were obtained using 5 different fields of view (4×4 cm, 6×6 cm, 8×8 cm, 10×10 cm and 10×5 cm), 2 types of rotation steps (180° and 360°) and 2 scanning resolutions (standard and high). Image analysis software was used to assess the trabecular bone microstructural parameters (number, thickness and spacing). All parameters were measured twice by one trained observer. Intraclass correlation coefficients showed high intraobserver repeatability (intraclass correlation coefficient, 0.95–0.97) in all parameters across all tested scan parameters. Trabecular bone microstructural measurements varied significantly, especially in smaller fields of view (p = 0.001). There was no significant difference in the trabecular parameters when using different resolutions (number, p = 0.988; thickness, p = 0.960; spacing, p = 0.831) and rotation steps (number, p = 1.000; thickness, p = 0.954; spacing, p = 0.759). The scan field of view significantly influences the trabecular bone microstructure measurements. Rotation steps (180° or 360°) and resolution (standard or high) selections are not relevant.
PMCID: PMC3853518  PMID: 24132024
cone beam CT; trabecular bone; diagnostic imaging; scanning parameter
12.  Field-derived Schistosoma mansoni and Biomphalaria pfeifferi in Kenya: a compatible association characterized by lack of strong local adaptation, and presence of some snails able to persistently produce cercariae for over a year 
Parasites & Vectors  2014;7:533.
Schistosoma mansoni is widely distributed in sub-Saharan Africa with Biomphalaria pfeifferi being its most widespread and important snail intermediate host. Few studies have examined the compatibility of field-derived B. pfeifferi snails with S. mansoni miracidia derived from human hosts. We investigated compatibility (as defined by shedding of cercariae following exposure to miracidia) of two isolates of S. mansoni from school children from Asao (western Kenya) and Mwea (central Kenya) with B. pfeifferi collected directly from Asao stream or the Mwea rice fields.
We exposed snails from both regions to four different doses of miracidia (1, 5, 10 and 25) from sympatric or allopatric S. mansoni, and maintained them in a shaded, screened out-of-doors rearing facility in Kisian, in western Kenya. Both snail survival and the number of snails that became infected were monitored weekly. This was done for 25 weeks post-exposure (PE). Those infected snails which survived beyond this period were monitored until they all died.
Although overall survival of Mwea snails maintained in western Kenya was generally low, both sympatric and allopatric combinations of parasites and snails exhibited high compatibility (approximately 50% at a dose of one miracidium per snail), with an increase in infection rates as the miracidial dose was increased (P < 0.002). Schistosomes were no more compatible with sympatric than allopatric snails, nor were snails less compatible with sympatric than allopatric schistosomes. Snail mortality increased significantly with dose of miracidia (P < 0.05). Approximately 3% of Asao snails exposed to a low dose of sympatric miracidia (1 or 5) continued to shed cercariae for as long as 58 weeks post exposure.
There were no significant local adaptation effects for either schistosomes or snails. Also, the existence of “super-survivor” snails is noteworthy for its implications for current control initiatives that mostly rely on mass drug administration (MDA). Long-term shedders could provide an ongoing source of cercariae to initiate human infections for many months, suggesting care is required in considering how human MDA treatments are timed. Future control programs should incorporate means to eliminate infected snails to complement chemotherapy interventions in controlling schistosomiasis.
Electronic supplementary material
The online version of this article (doi:10.1186/s13071-014-0533-3) contains supplementary material, which is available to authorized users.
PMCID: PMC4253995  PMID: 25425455
Schistosomiasis; Host-parasite interactions; Compatibility; Biomphalaria pfeifferi; Schistosoma mansoni; Super-sursvivor snails; Local adaptation; Parasite transmission
13.  No Apparent Reduction in Schistosome Burden or Genetic Diversity Following Four Years of School-Based Mass Drug Administration in Mwea, Central Kenya, a Heavy Transmission Area 
Schistosomiasis is a debilitating neglected tropical disease that infects over 200 million people worldwide. To combat this disease, in 2012, the World Health Organization announced a goal of reducing and eliminating transmission of schistosomes. Current control focuses primarily on mass drug administration (MDA). Therefore, we monitored transmission of Schistosoma mansoni via fecal egg counts and genetic markers in a typical school based MDA setting to ascertain the actual impacts of MDA on the targeted schistosome population.
For 4 years, we followed 67 children enrolled in a MDA program in Kenya. Infection status and egg counts were measured each year prior to treatment. For 15 of these children, for which there was no evidence of acquired resistance, meaning they became re-infected following each treatment, we collected microsatellite genotype data from schistosomes passed in fecal samples as a representation of the force of transmission between drug treatments. We genotyped a total of 4938 parasites from these children, with an average of 329.2 parasites per child for the entire study, and an average of 82.3 parasites per child per annual examination. We compared prevalence, egg counts, and genetic measures including allelic richness, gene diversity (expected heterozygosity), adult worm burdens and effective number of breeders among time points to search for evidence for a change in transmission or schistosome populations during the MDA program.
We found no evidence of reduced transmission or schistosome population decline over the course of the program. Although prevalence declined in the 67 children as it did in the overall program, reinfection rates were high, and for the 15 children studied in detail, schistosome egg counts and estimated adult worm burdens did not decline between years 1 and 4, and genetic diversity increased over the course of drug treatment.
School based control programs undoubtedly improve the health of individuals; however, our data show that in an endemic area, such a program has had no obvious effect on reducing transmission or of significantly impacting the schistosome population as sampled by the children we studied in depth. Results like these, in combination with other sources of information, suggest more integrated approaches for interrupting transmission and significantly diminishing schistosome populations will be required to achieve sustainable control.
Author Summary
Schistosomiasis is a chronic and debilitating disease. Current control focuses primarily on mass drug administration (MDA). We monitored schistosome transmission via fecal egg counts and genetic markers in a school based MDA setting to learn how the intervention was influencing transmission or the targeted schistosome population. For 4 years, we followed 67 children enrolled in a MDA program in Kenya, and for 15 of these, we repeatedly acquired in depth genetic data regarding the schistosome populations they harbored. Although prevalence declined in the 67 children, we found no evidence of reduced egg counts/worm burdens in those that reacquired infections and genetic diversity of schistosomes increased over the course of treatment. Our data indicate that this school based MDA program had a strong benefit to individual health as fewer children were infected over time. However, this decline does not appear to be due to schistosome population reduction, and may be caused by either acquired resistance or behavioral changes of the children. In conclusion, control programs based on chemotherapy alone or based on only a subset of the population will need to be supplemented with additional approaches if we are to achieve the WHO goal of eliminating human schistosomiasis by 2025.
PMCID: PMC4191953  PMID: 25299057
14.  Praziquantel sensitivity of Kenyan Schistosoma mansoni isolates and the generation of a laboratory strain with reduced susceptibility to the drug 
•We isolated S. mansoni miracidia from 72 Kenyan adults and children.•We found no evidence of S. mansoni with reduced praziquantel sensitivity.•An S. mansoni lab isolate with reduced praziquantel sensitivity was established.•The potential for the emergence of praziquantel resistance remains.
Schistosomiasis is a neglected tropical disease caused by blood-dwelling flukes of the genus Schistosoma. While the disease may affect as many as 249 million people, treatment largely relies on a single drug, praziquantel. The near exclusive use of this drug for such a prevalent disease has led to concerns regarding the potential for drug resistance to arise and the effect this would have on affected populations. In this study, we use an in vitro assay of drug sensitivity to test the effect of praziquantel on miracidia hatched from eggs obtained from fecal samples of Kenyan adult car washers and sand harvesters as well as school children. Whereas in a previous study we found the car washers and sand harvesters to harbor Schistosoma mansoni with reduced praziquantel sensitivity, we found no evidence for the presence of such strains in any of the groups tested here. Using miracidia derived from seven car washers to infect snails, we used the shed cercariae to establish a strain of S. mansoni with significantly reduced praziquantel sensitivity in mice. This was achieved within 5 generations by administering increasing doses of praziquantel to the infected mice until the parasites could withstand a normally lethal dose. This result indicates that while the threat of praziquantel resistance may have diminished in the Kenyan populations tested here, there is a strong likelihood it could return if sufficient praziquantel pressure is applied.
PMCID: PMC4266778  PMID: 25516840
Praziquantel; Schistosoma; Drug resistance; Miracidia; Schistosomiasis; Helminths
15.  Outcome of metastatic breast cancer in selected women with or without deleterious BRCA mutations 
Clinical & experimental metastasis  2013;30(5):10.1007/s10585-013-9567-8.
The aim of this study was to compare the time-to progression and overall survival (OS) in patients with metastatic breast cancer (MBC) with and without deleterious BRCA1/2 mutations. 195 women with MBC who were referred for BRCA genetic testing between 1997 and 2011 were included in the study. Logistic regression models and Cox proportional hazards models were fit to determine the associations between clinical variables and outcomes. Of 195 women with MBC, 21 % (n = 41) were positive for BRCA1/2 mutations. The number of metastatic sites at the time of metastatic disease was not different between BRCA1 versus BRCA2 carriers versus non-carriers (P = 0.77). The site of first metastasis was visceral-only in 70 % of BRCA1 carriers compared to 9 % in BRCA2 carriers and 37 % in non-carriers (P = 0.001). Median follow-up was 2.8 years. BRCA non-carriers and BRCA2 carriers had a longer time-to progression and OS compared to BRCA1 carriers (median time-to progression = 1.3 vs. 0.9 vs. 0.7 years; P = 0.31, and median OS = 4.88 vs. 4.94 vs. 1.34 years; P = 0.0065). In a multivariate model, no association was identified between BRCA positivity and time-to-event outcomes (P > 0.28). In addition, patients with triple-negative MBC carried a poorer prognosis irrespective of their BRCA status (P = 0.058 and P = 0.15 for the interaction term of BRCA status and triple-negative for time-to progression and OS, respectively). Our data indicate that BRCA1 carriers diagnosed with MBC have worse outcomes compared to BRCA2 carriers and non-carriers. However, the differences in outcome did not reach statistical significance likely due to small sample sizes.
PMCID: PMC3857726  PMID: 23370825
BRCA mutation; Chemotherapy; Metastatic breast cancer; Recurrence; Survival
16.  Diagnostic imaging of trabecular bone microstructure for oral implants: a literature review 
Dentomaxillofacial Radiology  2013;42(3):20120075.
Several dental implant studies have reported that radiographic evaluation of bone quality can aid in reducing implant failure. Bone quality is assessed in terms of its quantity, density, trabecular characteristics and cells. Current imaging modalities vary widely in their efficiency in assessing trabecular structures, especially in a clinical setting. Most are very costly, require an extensive scanning procedure coupled with a high radiation dose and are only partially suitable for patient use. This review examines the current literature regarding diagnostic imaging assessment of trabecular microstructure prior to oral implant placement and suggests cone beam CT as a method of choice for evaluating trabecular bone microstructure.
PMCID: PMC3667540  PMID: 23420864
dental implants; cone beam CT; microstructure
17.  Influence of cone beam CT scanning parameters on grey value measurements at an implant site 
Dentomaxillofacial Radiology  2013;42(3):79884780.
The aim of this study was to determine the grey value variation at the implant site with different scan settings, including field of view (FOV), spatial resolution, number of projections, exposure time and dose selections in two cone beam CT (CBCT) systems and to compare the results with those obtained from a multislice CT system.
A partially edentulous human mandibular cadaver was scanned by three CT modalities: multislice CT (MSCT) (Philips, Best, the Netherlands), and two CBCT systems: (Accuitomo 170®, Morita, Japan) and (NewTom 5G®, QR, Verona, Italy). Using different scan settings 36 and 24 scans were obtained from the Accuitomo and the NewTom, respectively. The scans were converted to digital imaging and communications in medicine 3 format. The analysis of the data was performed using 3Diagnosys® software (v. 3.1, 3diemme, Cantù, Italy) and Geomagic studio® 2012 (Morrisville, NC). On the MSCT scan, one probe designating the site for pre-operative implant placement was inserted. The inserted probe on MSCT was transformed to the same region on each CBCT scan using a volume-based three-dimensional registration algorithm. The mean voxel grey value of the region around the probe was derived separately for each CBCT. The influence of scanning parameters on the measured mean voxel grey values was assessed.
Grey values in both CBCT systems significantly deviated from Hounsfield unit values measured with MSCT (p = 0.0001). In both CBCT systems, scan FOV and spatial resolution selections had a statistically significant influence on grey value measurements (p = 0.0001). The number of projections selection had a statistically significant influence in the Accuitomo system (p = 0.0001) while exposure time and dose selections had no statistically significant influence on grey value measurements in the NewTom (p = 0.43 and p = 0.37, respectively).
Grey-level values from CBCT images are influenced by device and scanning settings.
PMCID: PMC3667541  PMID: 22933535
cone beam CT; registration; grey values; bone density
18.  Multiplex PCR in Determination of Opiinae Parasitoids of Fruit Flies, Bactrocera sp., Infesting Star Fruit and Guava 
Malaysia is a tropical country that produces commercial fruits, including star fruits, Averrhoa carambola L. (Oxalidales: Oxalidaceae), and guavas, Psidium guajava L. (Myrtales: Myrtaceae). There is a high demand for these fruits, and they are planted for both local consumption and export purposes. Unfortunately, there has been a gradual reduction of these fruits, which has been shown to be related to fruit fly infestation, especially from the Bactrocera species. Most parasitic wasps (Hymenoptera: Braconidae: Opiinae) are known as parasitoids of fruit fly larvae. In this study, star fruits and guavas infested by fruit fry larvae were collected from the Malaysian Agricultural Research and Development Institute. The parasitized larvae were reared under laboratory conditions until the emergence of adult parasitoids. Multiplex PCR was performed to determine the braconid species using two mitochondrial DNA markers, namely cytochrome oxidase subunit I and cytochrome b. Two benefits of using multiplex PCR are the targeted bands can be amplified simultaneously using the same reaction and the identification process of the braconid species can be done accurately and rapidly. The species of fruit flies were confirmed using the COI marker. The results obtained from our study show that Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), Fopius arisanus (Sonan), and Pysttalia incisi (Silvestri) were parasitoids associated with Bactrocera carambolae (Drew and Hancock) (Diptera: Tephritidae) infested star fruits. Fopius arisanus was also the parasitoid associated with Bactrocera papayae (Drew and Hancock) infested guavas. Maximum parsimony was been constructed in Opiinae species to compare tree resolution between these two genes in differentiating among closely related species. The confirmation of the relationship between braconids and fruit fly species is very important, recognized as preliminary data, and highly necessary in biological control programs.
PMCID: PMC4199358  PMID: 25373154
biological control programs; braconids; parasitic wasps
19.  Non-Invasive Sampling of Schistosomes from Humans Requires Correcting for Family Structure 
For ethical and logistical reasons, population-genetic studies of parasites often rely on the non-invasive sampling of offspring shed from their definitive hosts. However, if the sampled offspring are naturally derived from a small number of parents, then the strong family structure can result in biased population-level estimates of genetic parameters, particularly if reproductive output is skewed. Here, we document and correct for the strong family structure present within schistosome offspring (miracidia) that were collected non-invasively from humans in western Kenya. By genotyping 2,424 miracidia from 12 patients at 12 microsatellite loci and using a sibship clustering program, we found that the samples contained large numbers of siblings. Furthermore, reproductive success of the breeding schistosomes was skewed, creating differential representation of each family in the offspring pool. After removing the family structure with an iterative jacknifing procedure, we demonstrated that the presence of relatives led to inflated estimates of genetic differentiation and linkage disequilibrium, and downwardly-biased estimates of inbreeding coefficients (FIS). For example, correcting for family structure yielded estimates of FST among patients that were 27 times lower than estimates from the uncorrected samples. These biased estimates would cause one to draw false conclusions regarding these parameters in the adult population. We also found from our analyses that estimates of the number of full sibling families and other genetic parameters of samples of miracidia were highly intercorrelated but are not correlated with estimates of worm burden obtained via egg counting (Kato-Katz). Whether genetic methods or the traditional Kato-Katz estimator provide a better estimate of actual number of adult worms remains to be seen. This study illustrates that family structure must be explicitly accounted for when using offspring samples to estimate the genetic parameters of adult parasite populations.
Author Summary
Genetic epidemiology uses genetic data to uncover patterns of disease processes. To acquire data for these analyses, individual pathogens are collected and scored at genetic markers, and the resultant data are analyzed to infer biological patterns about the pathogen populations. In lieu of invasive sampling of adult pathogens in humans, researchers have relied on non-invasive sampling of parasite offspring (often shed in fecal samples). One potential problem with this approach is that analyses using the offspring data will be biased because many of the offspring are related and family sizes are likely to be unequal. We show that this sampling issue is relevant in a natural transmission zone in western Kenya and that it yields biases in three important parameters: genetic differentiation, inbreeding coefficients, and estimates of the amount of non-random association between loci (linkage disequilibrium). We also develop a method to remove these biases by removing the sibling structure present in the dataset. Finally, we suggest that our measure of family number, as well as other genetic measures, may be useful measures of the worm burdens in patients.
PMCID: PMC3777896  PMID: 24069499
20.  Polymorphism associated with the Schistosoma mansoni tetraspanin-2 gene 
International journal for parasitology  2011;41(12):1249-1252.
A vaccine against schistosomiasis would contribute significantly to reducing the 3-70 million disability-adjusted life years lost annually to the disease. Towards this end, inoculation with the large extracellular loop (EC-2) of Schistosoma mansoni tetraspanin-2 protein (Sm-TSP-2) has proved effective in reducing worm and egg burdens in S. mansoni-infected mice. The EC-2 loop of S. japonicum TSP-2, however, has been found to be highly polymorphic, perhaps diminishing the likelihood that this antigen can be used for vaccination against this species. Here, we examine polymorphism of the EC-2 of Sm-TSP-2 in genetically unique worms derived from six individuals from Kisumu, Kenya.
PMCID: PMC3188324  PMID: 21889508
Schistosoma; Polymorphism; Tetraspanin-2; Genotype; Kozak sequence; Vaccine
21.  Effect of novel water soluble curcumin derivative on experimental type- 1 diabetes mellitus (short term study) 
Diabetes mellitus type 1 is an autoimmune disorder caused by lymphocytic infiltration and beta cells destruction. Curcumin has been identified as a potent inducer of heme-oxygenase-1 (HO-1), a redoxsensitive inducible protein that provides protection against various forms of stress. A novel water soluble curcumin derivative (NCD) has been developed to overcome low in vivo bioavailability of curcumin. The aim of the present work is to evaluate the anti diabetic effects of the “NCD” and its effects on diabetes-induced ROS generation and lipid peroxidation in experimental type- 1 diabetes mellitus. We also examine whether the up regulation of HO-1 accompanied by increased HO activity mediates these antidiabetic and anti oxidant actions.
Materials and methods
Rats were divided into control group, control group receiving curcumin derivative, diabetic group, diabetic group receiving curcumin derivative and diabetic group receiving curcumin derivative and HO inhibitor ZnPP. Type-1 diabetes was induced by intraperitoneal injection of streptozotocin. Curcumin derivative was given orally for 45 days. At the planned sacrification time (after 45 days), fasting blood samples were withdrawn for estimation of plasma glucose, plasma insulin and lipid profile . Animals were sacrificed; pancreas, aorta and liver were excised for the heme oxygenase - 1 expression, activity and malondialdehyde estimation.
NCD supplementation to diabetic rats significantly lowered the plasma glucose by 27.5% and increased plasma insulin by 66.67%. On the other hand, the mean plasma glucose level in the control group showed no significant difference compared to the control group receiving the oral NCD whereas, NCD supplementation to the control rats significantly increased the plasma insulin by 47.13% compared to the control. NCD decreased total cholesterol, triglycerides, LDL cholesterol and increased HDL cholesterol levels. Also, it decreased lipid peroxides (malondialdehyde) in the pancreas, aorta and liver.
The (NCD) by its small dose possesses antidiabetic actions and that heme oxygenase induction seems to play an important role in its anti-diabetic effects. NCD also improves the lipid profile and oxidative status directly, proved by decreasing lipid peroxides (malondialdehyde) in pancreas, liver & aorta. The new water soluble curcumin derivative still retains the essential potencies of natural curcumin.
PMCID: PMC3533893  PMID: 22762693
Diabetes Type 1; Heme oxygenase −1; Curcumin; Insulin secretion; Oxidative stress
22.  Diffuse Large B-Cell Lymphoma Transformed from Mucosa-Associated Lymphoid Tissue Lymphoma Arising in a Female Urethra Treated with Rituximab for the First Time 
Case Reports in Oncology  2012;5(2):238-245.
A 30-year-old female patient presented to the gynecology clinic with a small (painless) swelling at the urethral orifice. She underwent surgical excision of the lesion. Pathological examination revealed non-Hodgkin's lymphoma of diffuse large B-cell type and mucosa-associated lymphoid tissue type, stage IE. The patient refused radiotherapy. Accordingly, we started CHOP-R chemotherapy. She received a total of 6 cycles of CHOP and 8 cycles of rituximab. Patient follow-up was done 3 months later through CT scan and cytoscopy confirming the complete remission. The patient has been disease-free for 4 years. We reviewed 26 cases of this rare entity reported previously.
PMCID: PMC3369256  PMID: 22679430
Urethra; NHL; Chemotherapy; Malignant lymphoma; Female urethra; CHOP-R chemotherapy
23.  (E)-2-(3-Cinnamoyl­thio­ureido)acetic acid dimethyl sulfoxide disolvate 
In the title compound, C12H12N2O3S·2C2H6OS, the acetic acid and cinnamoyl groups adopt Z and E configurations, respectively, with respect to the thio group about the C—N bonds. The components of the asymmetric unit are connected by N—H⋯O and O—H⋯O hydrogen bonds and in the crystal weak inter­molecular C—H⋯O and C—H⋯S hydrogen bonds further connect the components into chains along the b axis. In the main mol­ecule, an intra­molecular N—H⋯O hydrogen bond is also present.
PMCID: PMC3201284  PMID: 22058788
24.  3-(3-Meth­oxy­benzo­yl)-1,1-diphenyl­thio­urea 
The thiono and carbonyl groups in the title compound, C21H18N2O2S, adopt an anti disposition with respect to the central C—N bond. The diphenyl­amine rings are twisted relative to each other by a dihedral angle of 82.55 (10)°. The 3-meth­oxy­benzoyl fragment is twisted relative to one of the diphenyl­amine rings, forming a dihedral angle of 74.04 (9)°. In the crystal, pairs of inter­molecular N—H⋯S hydrogen bonds link the mol­ecules into centrosymmetric dimers, forming columns parallel to the a axis.
PMCID: PMC3212333  PMID: 22090990
25.  1,1-Dibenzyl-3-(4-fluoro­benzo­yl)thio­urea 
In the title compound, C22H19FN2OS, the 2-fluoro­benzoyl group adopts a trans conformation with respect to the thiono S atom across the N—C bond. In the crystal, inter­molecular N—H⋯S, C—H⋯S and C—H⋯O hydrogen bonds link the mol­ecules, forming a two-dimensional network parallel to (101).
PMCID: PMC3213443  PMID: 22091022

Results 1-25 (63)