HLA-G regulatory variants and haplotypes with susceptibility to recurrent pregnancy loss.

HLA-G is a nonclassical Class I major histocompatibility complex (MHC) gene. This gene has a limited protein alteration that is produced by alternative splicing and can be important in the preservation of pregnancy. Recent findings suggest that alteration in HLA-G gene expression can lead to pregnancy failure, such as recurrent pregnancy loss (RPL). As the promoter SNPs of the gene may impact the HLA-G expression levels, the study of these SNPs is very important. In this study, for the promoter region of HLA-G gene in the case group (100 women with a history of two or more repeated miscarriages) and the control group (100 women with at least two successful pregnancies), PCR reaction was performed. Thereafter, PCR products were sequenced and the results were compared between the two groups. The results showed that -1573T>C and -1746C>A SNPs in the promoter of the HLA-G gene associated with RPL. The outcome of the haplotype analysis also showed that the association of two haplotypes, including H1 (ATCCAGGTACGCAA) and H2 (CTTCGAGAACGCAG) with RPL, is significant. The results showed that H1 is associated with a decreased and H2 is associated with an increased risk of RPL. These results indicate the importance of the HLA-G promoter SNPs in the pregnancy outcome. But to reach a more definite conclusion, subsequent studies on 3' UTR and other positions with polymorphism in the 5' UTR regions larger samples are necessary.

Casein kinase 1-epsilon deletion increases mu opioid receptor-dependent behaviors and binge eating1.

Genetic and pharmacological studies indicate that casein kinase 1 epsilon (Csnk1e) contributes to psychostimulant, opioid, and ethanol motivated behaviors. We previously used pharmacological inhibition to demonstrate that Csnk1e negatively regulates the locomotor stimulant properties of opioids and psychostimulants. Here, we tested the hypothesis that Csnk1e negatively regulates opioid and psychostimulant reward using genetic inhibition and the conditioned place preference assay in Csnk1e knockout mice. Similar to pharmacological inhibition, Csnk1e knockout mice showed enhanced opioid-induced locomotor activity with the mu opioid receptor agonist fentanyl (0.2 mg/kg i.p.) as well as enhanced sensitivity to low-dose fentanyl reward (0.05 mg/kg). Interestingly, female knockout mice also showed a markedly greater escalation in consumption of sweetened palatable food - a behavioral pattern consistent with binge eating that also depends on mu opioid receptor activation. No difference was observed in fentanyl analgesia in the 52.5°C hot plate assay (0-0.4 mg/kg), naloxone conditioned place aversion (4 mg/kg), or methamphetamine conditioned place preference (0-4 mg/kg). To identify molecular adaptations associated with increased drug and food behaviors in knockout mice, we completed transcriptome analysis via mRNA sequencing of the striatum. Enrichment analysis identified terms associated with myelination and axon guidance and pathway analysis identified a differentially expressed gene set predicted to be regulated by the Wnt signaling transcription factor, Tcf7l2. To summarize, Csnk1e deletion increased mu opioid receptor-dependent behaviors, supporting previous studies indicating an endogenous negative regulatory role of Csnk1e in opioid behavior.

RNA-binding proteins, neural development and the addictions.

Transcriptional and post-transcriptional regulation of gene expression defines the neurobiological mechanisms that bridge genetic and environmental risk factors with neurobehavioral dysfunction underlying the addictions. More than 1000 genes in the eukaryotic genome code for multifunctional RNA-binding proteins (RBPs) that can regulate all levels of RNA biogenesis. More than 50% of these RBPs are expressed in the brain where they regulate alternative splicing, transport, localization, stability and translation of RNAs during development and adulthood. Dysfunction of RBPs can exert global effects on their targetomes that underlie neurodegenerative disorders such as Alzheimer's and Parkinson's diseases as well as neurodevelopmental disorders, including autism and schizophrenia. Here, we consider the evidence that RBPs influence key molecular targets, neurodevelopment, synaptic plasticity and neurobehavioral dysfunction underlying the addictions. Increasingly well-powered genome-wide association studies in humans and mammalian model organisms combined with ever more precise transcriptomic and proteomic approaches will continue to uncover novel and possibly selective roles for RBPs in the addictions. Key challenges include identifying the biological functions of the dynamic RBP targetomes from specific cell types throughout subcellular space (e.g. the nuclear spliceome vs. the synaptic translatome) and time and manipulating RBP programs through post-transcriptional modifications to prevent or reverse aberrant neurodevelopment and plasticity underlying the addictions.

Dynamic contrast-enhanced MRI for differentiation of major salivary glands neoplasms, a 3-T MRI study.

OBJECTIVES: Pre-operative differentiation of salivary gland neoplasms is of great importance. This study was designed to evaluate the use of dynamic contrast-enhanced MRI (DCE-MRI) for differentiation between malignant, Warthin and benign non-Warthin (BNW) neoplasms of major salivary glands. METHODS: 46 major salivary gland tumours (SGTs) underwent pre-operative DCE-MRI. Post-surgical histopathological evaluation showed 30 BNW, 6 Warthin and 10 malignant tumours. Time-signal intensity curves (TICs) were categorized as (a) Tpeak >43 s and washout ratio at 180 s (WR180) <4.6%; (b) Tpeak <43 s and WR >22%; (c) Tpeak >43 s and WR180 = 4.6-22.0% RESULTS: Accuracy of Tpeak was 98.9% for differentiation between BNW and Warthin tumours, 83.7% between BNW and malignant and 80% between malignant and Warthin tumours. All Warthin tumours showed Tpeak ≤43 s, while one BNW had Tpeak <43 s. A Tpeak <63.5 s differentiated 8/10 (80%) malignant tumours from BNW tumours, whereas 4/30 of BNW tumours had a Tpeak <63.5 s. Two malignant tumours had Tpeak <43 s. WR180 had an accuracy of 100% for differentiation between Warthin and BNW tumours, 87.3% between BNW and malignant, and 93.3% between Warthin and malignant tumours. 29 (96.7%) BNW tumours had a washout <4.60%, while 8 (80%) malignant tumours had a washout >4.60%. All Warthin tumours had a WR180 >22%, while two malignant tumours had a WR180 >22%. 29/30 of BNW tumours demonstrated TIC curve Type A and 1 tumour demonstrated Type C. 6/10 of malignant tumours had TIC Type C, 2 had TIC Type A and 2 Type B. All Warthin tumours were categorized as Type B. CONCLUSIONS: This study showed that DCE-MRI could be helpful in pre-operative differentiation of SGTs; especially for discrimination between Warthin and BNW tumours.

Association between MIF gene variation and Meniere's disease.

Several pieces of evidence support the involvement of immune system in Menière's disease (MD). Macrophage migration inhibitory factor (MIF) plays a key role in immune-mediated reactions. Several studies have shown an association between MIF gene polymorphisms and susceptibility to various inflammatory and autoimmune disorders. The aim of this study was to explore the association between MIF-173 G/C polymorphism and MD in an Iranian population. In this case-control association study, MD cases (N = 72) were recruited and were comprised of definitive MD (N = 58) and probable MD (N = 14) subjects. Normal healthy subjects (N = 100) were also included. Genotyping for MIF-173 G/C polymorphism was carried out using PCR-RFLP technique. There was a significant increase in genotype GG in patients with MD compared with the control group. (GG vs. GC + CC, P = 0.02, OR = 2.08, 95% CI: 1.02-4.3). This was more significant when definitive MD was stratified and compared with the controls (GG vs. GC + CC, P = 0.009, OR = 2.6, 95% CI = 1.19-6.18). This study's result indicates the potential role of MIF in MD of which further evaluation is required. Also, the more significant association between MIF gene polymorphism and definitive MD designates the involvement of specific pathogenic mechanisms which may be considered as a marker for diagnosis.

Association between the functional polymorphism C-159T in the CD14 promoter gene and nasal polyposis: potential role in asthma.

BACKGROUND: Nasal polyposis (NP) is a chronic inflammatory disease that is frequently associated with allergy and asthma. Corticosteroid therapy and surgical removal of polyps are the 2 most common treatment strategies for NP. Various allergic and inflammatory mediators are thought to play a major role in the pathophysiology of this disorder. The CD14 gene is located on chromosome 5q31-32, which is considered a critical region for several allergic and atopic diseases, including asthma. Consequently, variations in CD14 could have functional effects on the etiology and severity of allergy and asthma. The aim of this study was to investigate the association between the polymorphism C-159T in the CD14 gene of patients with NP and controls. METHODS: The study population comprised 106 patients with NP diagnosed based on computed tomography scan of the paranasal sinus, endoscopy, and histological examination. Findings were compared with those from 87 controls. The frequency of C-159T was determined using polymerase chain reaction-restriction fragment length polymorphism analysis. DNA was extracted using the salting out technique. RESULTS: A significant association was observed between C-159T and NP (P = .04). Patients with the CC genotype at position -159 of the CD14 promoter region had an increased risk of asthma (OR, 3.83, 95% CI, 0.99-13.91; P < .02). However, we did not find an association between the distribution of C-159T and serum immunoglobulin E level. CONCLUSIONS: A genetic variation in the CD14 promoter might play a role in the pathogenesis of NP and in the incidence of asthma.

The presence of amorpha-4, 11-diene synthase, a key enzyme in artemisinin production in ten Artemisia species.

BACKGROUND AND THE PURPOSE OF THE STUDY: Artemisinin is one of the most effective medicine against malaria, which is produced naturally by Artemisia annua in low yield. It is produced in a metabolic pathway, in which several genes and gene products are involved. One of the key genes in this pathway is am1, which encodes amorpha-4, 11-diene synthase (ADS), a key enzyme in artemisinin biosynthesis pathway. The aim of this study was to determine the presence of this gene in ten Artemisia species in order to increase the yield of production of Artemisinin. METHODS: The experiments were carried out using PCR. Specific primers were designed based on the published am1 gene sequence obtained from A. annua (NCBI, accession number AF327527). RESULTS: The amplification of this gene by the specific primers was considered as a positive sign for the potentiality of artemisinin production. Since the entire am1 gene was not amplified in any of the 10 species used, four parts of the gene, essential in ADS enzyme function, corresponding to a) pair site of Arg10-Pro12 in the first 100 amino acids, b) aspartate rich motif (DDXXD), c) active site final lid and d) active site including farnesyl diphosphate (FDP) ionization sites and catalytic site in the ADS enzyme, were investigated. MAJOR CONCLUSION: The sequence corresponding to ADS active site was amplified only in A. annua, A. aucheri and A. chamaemelifolia. The negative results obtained with other species could be due to some sequence alteration, such as point mutations or INDELs. We propose A. aucheri and A. chamaemelifolia as two potential candidate species for further characterization, breeding and transferring am1 gene for artemisinin overproduction.

Infratemporal hydatid cyst: a case presenting with blindness.

OBJECTIVE: We report a very rare case of a hydatid cyst in the infratemporal fossa, causing visual loss over a 10-day period, which disappeared with rapid surgical and medical treatment. CASE REPORT: A 14-year-old girl presented with right exophthalmos and visual loss. Over a 10-day period, her visual acuity had decreased to detection of hand motion only, due to pressure on the optic nerve caused by a parapharyngeal cyst pressing through a inferior orbital fissure on the right side. A craniotomy had previously been performed for a right frontoparietal hydatid cyst. The patient had been treated intermittently with albendazole. The patient was primarily diagnosed with hydatid cyst, on the basis of her previous medical history and radiological findings, and underwent surgery. Three cysts were carefully removed from the right maxillary sinus, via a standard Caldwell-Luc approach, and the surgical area was irrigated with hypertonic saline. CONCLUSION: Infratemporal hydatidosis is very rarely reported in the world literature, although hydatid cysts are endemic in many countries, including Iran. We discuss the common presenting features, investigation and treatment options for infratemporal hydatosis. Constant evaluation of adjacent organs is necessary, with treatment as required, due to the propensity of hydatidosis to recur in essential organs. Immediate surgery is recommended, both to prevent the development of disease and to improve the prognosis.

Transient facial nerve baroparesis: case report.

We present a rare case of facial paralysis that was unusual not only in its causation but also in its rapidity of onset and recovery. We describe a rare case history of this accruing in 35 years old women traveling at the high altitude mountain road referred to ENT clinic with sudden symptoms of middle ear effusion and facial nerve paralysis. Patient had undergone medical systemic steroid treatment and after 3-4 weeks she had a good recovery of facial palsy with a minimum remnant of sensory neural hearing loss. Facial nerve paralysis resulting from a barotrauma of the middle ear is suggested. The correct diagnosis of this condition is important and unnecessary treatment should be avoided.

Nasal vestibular huge keratoacanthoma: an unusual site.

Keratoacanthoma (KA) is a rapidly growing, low-grade neoplasm of pilo-sebaceous and hair follicle units which most often appears on the sun-exposed skin of the middle aged and older persons with multiple or localized occurrence. This tumor is dome-shaped nodule with a central keratinous plug. The etiology of this tumor is not obvious. Exposure to excessive sunlight is the most frequently noted responsible factor in the etiology of KA. About 80% of the tumors occur on the face. The histological features of the KA are often very similar to those of a cutaneous squamous cell carcinoma; however, the tumor structure usually provides a basis for their difference. There are many unusual cases of keratoacanthoma reported regarding site, size or other specifications. In this study, we excised a mass of nasal vestibule, a site far away sun-exposure. To our knowledge, this is the first case of nasal vestibular keratoacanthoma. For a clinician and a pathologist it is important to consider a benign lesion like Keratoacanthoma (KA) in the differential diagnosis of ulcerated nasal lesions and pay attention to differ it from Squamous Cell Carcinoma (SCC) which has a different and aggressive management.

Auricular leishmaniasis mimicking squamous cell carcinoma.

OBJECTIVES: We report a rare case of auricular involvement by leishmaniasis, in order to demonstrate the importance of thorough investigation of cutaneous head and neck lesions, and also the importance of inclusion of infections such as leishmaniasis in the differential diagnosis of auricular lesions, especially in endemic areas. CASE HISTORY: A 42-year-old man with multiple lesions on his head, neck and hands was referred to our centre. He had the following lesions: a painful, crusted, 8 x 8 cm plaque with indurated margins on the left parotid region and auricle; a red papule on the right temporal region; an ulcerative lesion on the skin overlying the proximal interphalangeal joint of the fifth finger of the right hand; and a bluish papule on the neck. Although histopathological examination of the Geimsa-stained specimen was misleading, a direct smear prepared from biopsies showed amastigotes, and therapy resulted in complete recovery. CONCLUSION: Leishmaniasis can be both under- or over-diagnosed. Especially in endemic areas, parasitic causes of chronic infections should always be kept in mind.

Sci-Thurs PM: Delivery-01: Sliding Window IMRT: Uncertainties of the leading edge and plateau of the beam profile.

The aim of this work is to analyze the dependence of the dose profile uncertainties for the sliding window IMRT (SW-IMRT) beams under the condition of an extreme dose rate (DR) and leaf velocity (LV). The deviations of the edges and plateau for the beam profiles of small number of MUs delivered using the dynamic MLC were studied. Field sizes with lengths of 5 and 10 cm were irradiated by photon beams of 2-8 MU/beam, DR = 100-600 MU/min and LS = 1-5 cm/s. Kodak TL radiographic films were used in the measurement. The photon beams (6 and 15 MV) were produced by a Varian 21EX Linac with a 120-leaf MLC. It is found that the MLC cannot keep the leaves moving with a proper speed continuously under a stable DR when beam of small MUs are irradiated. For example, the dynamic MLC needs 1.2 s and 12 MU to irradiate a field of 5 cm length with 2 MU using DR = 600 MU/min and LC = 5 cm/s. The plateau of the beam profile has several sinusoidal periods of about 150 ms. The magnitude of the plateau uncertainties was about 7% and 15% for the dose of the beam with DR = 400 and 600 MU/min (2 MU/beam), respectively. It is concluded that SW-IMRT beams of more than 10 MUs, delivered with 1 cm/s ⩽ LV ⩽ 5 cm/s and 100 MU/min ⩽ DR ⩽ 600 MU/min, have a good agreement between the delivered and planned dose profiles.

Simplified sperm retrieval and intracytoplasmic sperm injection in patients with azoospermia.

OBJECTIVE: To evaluate the rate of recovery of spermatozoa from the epididymis using a percutaneous aspiration technique and to assess the fertilization rate following intracytoplasmic sperm injection (ICSI). PATIENTS AND METHODS: Forty-two patients with azoospermia underwent a total of 46 treatment cycles of in vitro fertilization (IVF) and ICSI. The sperm used for ICSI was retrieved percutaneously by fine-needle aspiration and syringe suction (percutaneous epididymal sperm aspiration, PESA) from the epididymis in 28 patients (mean age 34.9 years), over 32 cycles. Six patients underwent microsurgical sperm aspiration (MESA) and in the remaining eight patients, neither percutaneous aspiration nor MESA yielded suitable sperm and spermatozoa extracted from testicular biopsy were used. RESULTS: A total of 362 oocytes were collected and of those, 286 (79%) were subjected to ICSI. Of the injected oocytes, 49 (17.2%) were damaged, 138 (48.3%) achieved normal fertilization and, of those, 112 (81.2%) cleaved. A total of 67 embryos were transferred and 18 more were suitable for cryopreservation. Of the 25 cycles with embryo transfer, eight resulted in a pregnancy and of these, one miscarried. The pregnancy rate was 25% per cycle and 32% per embryo transfer. The implantation rate was 12%. CONCLUSIONS: This extensive series of PESA/ICSI cycles indicates that PESA can be used successfully to retrieve spermatozoa in patients with azoospermia. The technique is simple, cost-effective and is associated with fewer complications compared to an open microsurgical procedure.

The time of appearance and disappearance of fetal DNA from the maternal circulation.

A single copy Y-chromosome DNA sequence was amplified using the polymerase chain reaction (PCR) from the peripheral blood of 30 women who had achieved a pregnancy through an in vitro fertilization (IVF) programme. The time of conception was known precisely and was confirmed by serial ultrasound scans. Conceptions were dated as the number of weeks after fertilization plus 2, to give a time equivalent to the obstetric menstrual dating of the pregnancy (LMP). Y-chromosome-specific DNA was detected in all pregnancies with a male fetus (18/30). The earliest detection was at 4 weeks and 5 days, and the latest at 7 weeks and 1 day. Y-chromosome-specific sequences were no longer detected in any of the male pregnancies 8 weeks after delivery. No Y-chromosome sequences were detected in any of the pregnancies where only female babies were delivered. This demonstrates that fetal DNA appears in the maternal circulation early in the first trimester, that it can be identified in all pregnancies tested by 7 weeks, that it continues to be present throughout pregnancy, and that it has been cleared from the maternal circulation 2 months after parturition. Early non-invasive prenatal diagnosis for aneuploidies and inherited disorders will be possible in all pregnancies if fetal cells can be isolated free from maternal contamination (or identified accurately in the presence of maternal cells) without problems of contamination from previous pregnancies.

Intramural pregnancy following difficult embryo transfer.

We report an intramural pregnancy following a difficult embryo transfer in a 31 year-old woman, having in-vitro fertilization and embryo transfer for tubal factor infertility. The creation of a 'false passage' at a previous instrumentation of the cervix may be implicated in the ectopic placement of embryos.

Experience with subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI) on unfertilized aged human oocytes.

OBJECTIVE: The aim of this study was to assess the fertilizability of unfertilized aged human oocytes from failed in vitro fertilization (IVF) cycles using SUZI and ICSI. METHODS: A total of 363 oocytes which showed no fertilization after conventional IVF was subjected to assisted fertilization using SUZI or ICSI. The microinjected oocytes which were derived from 72 patients undergoing their first IVF treatment had an intact polar body and no signs of degeneration. SUZI was carried out in 265 oocytes and ICSI in the remaining 98. RESULTS: Significantly more oocytes were damaged after ICSI (9 vs 0.3%, P < 0.01). Normal fertilization rates were higher at 24 hr in both groups and occurred more frequently after ICSI, although the difference did not reach statistical significance. Abnormal fertilization occurred significantly more often after SUZI at 48 hr (P < 0.005), but not at 24 hr. Cleavage rates were significantly higher after ICSI (94.4 vs 57.1%, P < 0.025) at 24 hr, but this was not observed at 48 hr, although the ICSI group still showed better cleavage rates (33.3 vs 19.1%). There was no difference in embryo quality in either group. CONCLUSIONS: Our results indicate that micromanipulation rather than reinsemination should be carried out on unfertilized human oocytes from failed IVF attempts. Both techniques can be used to achieve fertilization which occurs more often after ICSI. However, the trauma from the former technique on the microinjected oocytes may impair the potential of the generated embryos to achieve pregnancy compared to SUZI. Prospective randomized trials are necessary to address the problem.