Cytokinesis, the final stage of cell division, bisects the cytoplasm into two daughter cells. In mitotic cells, this process depends on the activity of non-muscle myosin II (NMII), a family of actin-binding motor-proteins that participate in the formation of the cleavage furrow. The relevance of NMII for meiotic cell division, however, is poorly understood. The NMII family consists of three members, NMIIA, NMIIB, and NMIIC, containing different myosin heavy chains (MYH9, MYH10, and MYH14, respectively). We find that a single non-muscle myosin II, NMIIB, is required for meiotic cytokinesis in male but not female mice. Specifically, NMIIB-deficient spermatocytes exhibit cytokinetic failure in meiosis I, resulting in bi-nucleated secondary spermatocytes. Additionally, cytokinetic failure at meiosis II gives rise to bi-nucleated or even tetra-nucleated spermatids. These multi-nucleated spermatids fail to undergo normal differentiation, leading to male infertility. In spite of the presence of multiple non-muscle myosin II isoforms, we demonstrate that a single member, NMIIB, plays an essential and non-redundant role in cytokinesis during meiotic cell divisions of the male germline.
Spermatogenesis; Meiosis; Cytokinesis; NMIIB; MYH10; Mouse
Chemo-resistance is one of the key causal factors in cancer death and emerging evidences suggest that microRNAs (miRNAs) have critical roles in the regulation of chemo-sensitivity in cancers. Cervical cancer is one of the most common malignancies in women and insensitive to chemotherapy clinically.
The differentially expressed miRNAs in cervical squamous cell carcinoma tissues were screened by using a microarray platform (μParaflo Sanger miRBase release 13.0). The expression of miR-375 was determined by stem-loop RT–PCR using 23 clinical cervical cancer samples and 2 cervical cancer cell lines. We exogenously upregulated miR-375 expression in SiHa and Caski cells using a pre-miRNA lentiviral vector transfection and observed its impact on paclitaxel sensitivity using MTS. The cells that stably overexpressed miR-375 were subcutaneously injected into mice to determine tumour growth and chemo-sensitivity in vivo.
Twenty-one differentially expressed miRNAs were found by miRNA microarray between pro- and post-paclitaxel cervical cancer tissues. Of those, miR-375 showed consistent high expression levels across paclitaxel-treated cervical cells and tissues. Paclitaxel induced upregulated miR-375 expression in a clear dose-dependent manner. Forced overexpression of miR-375 in cervical cancer cells decreased paclitaxel sensitivity in vitro and in vivo.
Collectively, our results suggest that miR-375 might be a therapeutic target in paclitaxel-resistant cervical cancer.
miR-375; paclitaxel; chemo-resistance; cervical cancer
This was a post hoc analysis of patients with non-squamous histology from a phase III maintenance pemetrexed study in advanced non-small cell lung cancer (NSCLC).
Patients and methods
The six symptom items' [average symptom burden index (ASBI)] mean at baseline was calculated using the lung cancer symptom scale (LCSS). Low and high symptom burden (LSB, ASBI < 25; HSB, ASBI ≥ 25) and performance status (PS: 0, 1) subgroups were analyzed for treatment effect on progression-free survival (PFS) and overall survival (OS) using the Cox proportional hazard models adjusted for demographic/clinical factors.
Significantly longer PFS and OS for pemetrexed versus placebo occurred in LSB patients [PFS: median 5.1 versus 2.4 months, hazard ratio (HR) 0.49, P < 0.0001; OS: median 17.5 versus 11.0 months, HR 0.63, P = 0.0012] and PS 0 patients (PFS: median 5.5 versus 1.7 months, HR 0.36, P < 0.0001; OS: median 17.7 versus 10.3 months, HR 0.54, P = 0.0019). Significantly longer PFS, but not OS, occurred in HSB patients (median 3.7 versus 2.8 months, HR 0.50, P = 0.0033) and PS 1 patients (median 4.4 versus 2.8 months, HR 0.60, P = 0.0002).
ASBI and PS are associated with survival for non-squamous NSCLC patients, suggesting that maintenance pemetrexed is useful for LSB or PS 0 patients following induction.
lung cancer symptom scale; maintenance therapy; non-squamous NSCLC; patient-reported symptoms; pemetrexed; survival
To evaluate the accuracy of 64-slice CT angiography (CTA) compared with single photon emission CT (SPECT) myocardial perfusion imaging (MPI), which served as the reference standard, for the detection of functionally significant coronary artery disease (CAD).
141 consecutive patients (60±10 years, 101 men) were investigated with 64-slice CTA and SPECT MPI; a subset of 35 patients had additional invasive coronary angiography (ICA). The data from CTA and ICA were compared with those from MPI for both cut-offs of ≥50% and ≥70% stenosis, respectively.
The sensitivity, specificity, positive and negative predictive values, and accuracy of CTA, using a cut-off of ≥50% for significant stenosis, in detecting inducible perfusion defects on MPI were 96% [95% confidence interval (CI) 88–100%], 61% (95% CI 52–70%), 37% (95% CI 23–49%), 99% (95% CI 97–100%) and 68%, respectively, in patient-based analysis and 97% (95% CI 91–100%), 86% (95% CI 83–89%), 33% (95% CI 24–42%), 100% (95% CI 99–100%) and 87%, respectively, in vessel-based analysis. Applying a cut-off of ≥70% for significant stenosis, CTA yielded the following sensitivity, specificity, positive and negative predictive values, and accuracy for the detection of inducible MPI defects: by patient, 65% (95% CI 46–84%), 95% (95% CI 91–99%), 74% (95% CI 50–92%), 92% (95% CI 87–97%) and 89%, respectively; by vessel, 58% (95% CI 42–74%), 97% (95% CI 95–99%), 62% (95% CI 45–79%), 97% (95% CI 95–99%) and 95%, respectively.
64-slice CTA is a reliable tool to exclude functionally significant CAD when using a cut-off of ≥50% diameter stenosis. By contrast, a cut-off of ≥70% diameter narrowing is a strong predictor of ischaemia.
The aim of this retrospective study was to characterize MRI findings of synovial chondromatosis in the temporomandibular joint (TMJ) by correlation with their pathological findings.
22 patients with synovial chondromatosis in unilateral TMJ were referred for plain MRI prior to surgical management and pathological examinations. Parasagittal and coronal proton density-weighted imaging and T2 weighted imaging were performed for each case.
MRI demonstrated multiple chondroid nodules and joint effusion in all patients (100%) and amorphous iso-intensity signal tissues within expanded joint space and capsule in 19 patients (86.4%). On T2 weighted imaging, signs of low signal nodules within amorphous iso-intensity signal tissues were used to determine the presence of attached cartilaginous nodules in pathology, resulting in 100% sensitivity, 60% specificity and 90.9% accuracy. Signs of low and intermediate signal nodules within joint fluids were used to detect loose cartilaginous nodules and resulted in 80% sensitivity, 42.9% specificity and 68.2% accuracy.
MRI of synovial chondromatosis in TMJ was characterized by multiple chondroid nodules, joint effusion and amorphous iso-intensity signal tissues within the expanded space and capsule. The attached cartilaginous nodules in pathology were better recognized than the loose ones on MRI. Plain MRI was useful for clinical diagnosis of the disorder.
synovial chondromatosis; tempromandibular joint; magnetic resonance imaging
The aim of this study was to analyse 19 recurrent keratocystic odontogenic tumours (KCOTs).
19 patients with recurrent KCOTs were retrospectively analysed. These patients had been treated by either enucleation or a combination of enucleation and Carnoy's solution. The analyses covered major aspects of primary KCOT and/or recurrent KCOT identities, including patient profile, clinical features, histopathology, radiology, treatment and prognosis.
19 (7.4%) out of 257 primary KCOT cases recurred, with an average patient age of 30.5 years (age range 18–45 years). 15 lesions were in the mandible and the remaining 4 were in the maxilla. There were more unilocular than multilocular occurrences for the primary tumours, with a ratio of 2.2:1. These KCOTs were initially treated by simple enucleation (12 cases) or enucleation with Carnoy's solution (7 cases). After the initial surgery, 15 out of 19 (78.9%) recurred within 6 years, while 4 (21.1%) recurred after 6 years. Evidently, the recurrent lesion was involved with the roots of the teeth in three out of six cases whose teeth were preserved. In addition, the recurrent KCOTs had a tendency to be more multilocular or multifocal than the primary cases, with a unilocular-to-multilocular ratio of 1.1:1.
7.4% of primary KCOTs recurred within 6 years after initial treatment with either enucleation or a combination of enucleation and Carnoy's solution. The recurrent KCOTs were more likely to be multilocular or multifocal than the primary cases and often involved the teeth. The method of operation for these recurrent lesions would be considered as a more aggressive approach.
keratocystic odontogenic tumour; odontogenic keratocyst; enucleation; recurrence
The diverse transcriptional mechanisms governing cellular differentiation and development of mammalian tissue remains poorly understood. Here we report that TAF7L, a paralogue of TFIID subunit TAF7, is enriched in adipocytes and white fat tissue (WAT) in mouse. Depletion of TAF7L reduced adipocyte-specific gene expression, compromised adipocyte differentiation, and WAT development as well. Ectopic expression of TAF7L in myoblasts reprograms these muscle precursors into adipocytes upon induction. Genome-wide mRNA-seq expression profiling and ChIP-seq binding studies confirmed that TAF7L is required for activating adipocyte-specific genes via a dual mechanism wherein it interacts with PPARγ at enhancers and TBP/Pol II at core promoters. In vitro binding studies confirmed that TAF7L forms complexes with both TBP and PPARγ. These findings suggest that TAF7L plays an integral role in adipocyte gene expression by targeting enhancers as a cofactor for PPARγ and promoters as a component of the core transcriptional machinery.
The development of a single fertilized egg into a highly complex animal is determined by its genome, with a process called differential gene regulation exerting exquisite control over gene expression to ensure that various specialized cells are generated and that many types of tissue are produced. However, the mechanisms responsible for controlling gene expression and, therefore mammalian development, are poorly understood.
Researchers have developed a number of in vitro cell culture models to elucidate the details of differential gene regulation, and this approach has been used to characterize adipocytes—cells that store energy in the form of fat—for close to two decades. The formation of adipocytes, a process known as adipogenesis, has been extensively studied, but there remain major gaps in our knowledge: for example, the identities of many of the transcriptional regulators that are responsible for the differentiation of mesenchymal stem cells into adipocytes remain a mystery. This task is complicated by the fact that some of these regulators are involved in the differentiation of multiple cell lines, and that some of them also have multiple roles in the generation of a single cell type. In addition to being of fundamental interest, improving our knowledge of the properties and behavior of adipocytes is essential for tackling the increasing prevalence of obesity in the developed world.
Zhou et al. now report that TAF7L—a gene that was previously thought to be involved only in the production of sperm cells—has two roles in the differentiation of stem cells to form adipocytes. Using a combination of cellular, biochemical, genetic and genomic techniques, they show that TAF7L interacts with PPARγ, an important adipocyte transcriptional regulator at enhancer sites on the genome to increase the transcription of genes that are involved in adipogenesis. They also show that TAF7L interacts with a general transcription factor called TBP (short for TATA-binding protein) at promoter sequences, again to increase the expression of genes involved in adipogenesis. Moreover, they show that the expression of TAF7L in myoblasts—precursor cells that usually become muscle cells—can induce the formation of fat cells rather than muscle cells. Furthermore, mice lacking TAF7L are lean compared to their normal littermates. A clearer understanding of the underlying causes of fat cell formation could lead to the development of new approaches for the treatment of obesity and associated diseases.
ChIP-seq; RNA-seq; adipogenesis; C3H10T½; TAF7L; differentiation; Mouse
Piwi-interacting RNAs are a diverse class of small non-coding RNAs implicated in the silencing of transposable elements and the safeguarding of genome integrity. In mammals, male germ cells express two genetically and developmentally distinct populations of piRNAs at the pre-pachytene and pachytene stages of meiosis, respectively. Pre-pachytene piRNAs are mostly derived from retrotransposons and required for their silencing. In contrast, pachytene piRNAs originate from ∼3,000 genomic clusters, and their biogenesis and function remain enigmatic. Here, we report that conditional inactivation of the putative RNA helicase MOV10L1 in mouse spermatocytes produces a specific loss of pachytene piRNAs, significant accumulation of pachytene piRNA precursor transcripts, and unusual polar conglomeration of Piwi proteins with mitochondria. Pachytene piRNA–deficient spermatocytes progress through meiosis without derepression of LINE1 retrotransposons, but become arrested at the post-meiotic round spermatid stage with massive DNA damage. Our results demonstrate that MOV10L1 acts upstream of Piwi proteins in the primary processing of pachytene piRNAs and suggest that, distinct from pre-pachytene piRNAs, pachytene piRNAs fulfill a unique function in maintaining post-meiotic genome integrity.
Small non-coding RNAs play critical roles during development and in disease. The integrity of the germline genome is of paramount importance to the wellbeing of offspring and the survival of species. Piwi-interacting RNAs (piRNAs) are a class of small non-coding RNAs abundantly expressed in the gonad. Compared to microRNAs and small-interfering RNAs (siRNAs), the biogenesis and function of piRNAs remain poorly understood. Here we have identified MOV10L1, a putative RNA helicase, as a master regulator of piRNA biogenesis in mouse. We find that production of pachytene piRNAs requires MOV10L1. Blockade of pachytene piRNAs disrupts germ cell development and results in defects in post-meiotic genome integrity. Therefore, mutations in MOV10L1 and other piRNA pathway components may contribute to male infertility in humans.
CHTF18 (chromosome transmission fidelity factor 18) is an evolutionarily conserved subunit of the Replication Factor C-like complex, CTF18-RLC. CHTF18 is necessary for the faithful passage of chromosomes from one daughter cell to the next during mitosis in yeast, and it is crucial for germline development in the fruitfly. Previously, we showed that mouse Chtf18 is expressed throughout the germline, suggesting a role for CHTF18 in mammalian gametogenesis. To determine the role of CHTF18 in mammalian germ cell development, we derived mice carrying null and conditional mutations in the Chtf18 gene. Chtf18-null males exhibit 5-fold decreased sperm concentrations compared to wild-type controls, resulting in subfertility. Loss of Chtf18 results in impaired spermatogenesis; spermatogenic cells display abnormal morphology, and the stereotypical arrangement of cells within seminiferous tubules is perturbed. Meiotic recombination is defective and homologous chromosomes separate prematurely during prophase I. Repair of DNA double-strand breaks is delayed and incomplete; both RAD51 and γH2AX persist in prophase I. In addition, MLH1 foci are decreased in pachynema. These findings demonstrate essential roles for CHTF18 in mammalian spermatogenesis and meiosis, and suggest that CHTF18 may function during the double-strand break repair pathway to promote the formation of crossovers.
Meiosis is the specialized process of cell division during germ cell development that results in formation of eggs and sperm. Genetic exchange between maternal and paternal chromosomes occurs during meiosis in a process called homologous recombination, in which DNA double- strand breaks are made and then repaired to allow DNA crossovers to form. These are essential processes that keep homologous chromosomes joined until anaphase I and ensure proper chromosome segregation. Errors in meiotic recombination lead to chromosome mis-segregation and ultimately aneuploidy, an abnormal chromosome number. Although it is well known that defects in these processes contribute greatly to infertility, birth defects, and pregnancy loss in humans, their molecular basis is not well understood. We demonstrate here a Chtf18 mutant mouse that exhibits subfertility and defects in meiotic recombination. Specifically, DNA double-strand breaks are incompletely repaired, DNA crossovers are significantly decreased, and homologous chromosomes separate during prophase I in Chtf18-null males. Our findings suggest roles for CHTF18 in DNA double-strand break repair and crossover formation, functions in mammals not previously known.
The human motor cortex exhibits characteristic beta (15-30 Hz) and gamma oscillations (60-90 Hz), typically observed in the context of transient finger movement tasks. The functional significance of these oscillations, such as post-movement beta rebound (PMBR) and movement-related gamma synchrony (MRGS) remain unclear. Considerable animal and human non-invasive studies, however, suggest that the networks supporting these motor cortex oscillations depend critically on the inhibitory neurotransmitter γ-Aminobutyric acid (GABA). Despite such speculation, a direct relation between MEG measured motor cortex oscillatory power and frequency with resting GABA concentrations has not been demonstrated.
In the present study, motor cortical responses were measured from 9 healthy adults while they performed a cued button-press task using their right index finger. In each participant, PMBR and MRGS measures were obtained from time-frequency plots obtained from primary motor (MI) sources, localized using beamformer differential source localization. For each participant, complimentary magnetic resonance spectroscopy (MRS) GABA measures aligned to the motor hand knob of the left central sulcus were also obtained. GABA concentration was estimated as the ratio of the motor cortex GABA integral to a cortical reference NAA resonance at 2 ppm.
A significant linear relation was observed between MI GABA concentration and MRGS frequency (R2 = 0.46, p<0.05), with no association observed between GABA concentration and MRGS power. Conversely, a significant linear relation was observed between MI GABA concentration and PMBR power (R2 = 0.34, p<0.05), with no relation observed for GABA concentration and PMBR frequency. Finally, a significant negative linear relation between the participant’s age and MI gamma frequency was observed, such that older participants had a lower gamma frequency (R2 = 0.40, p < 0.05).
Present findings support a role for GABA in the generation and modulation of endogenous motor cortex rhythmic beta and gamma activity.
Magnetic Resonance Spectroscopy (MRS); γ-Aminobutyric acid (GABA); Event-related desynchrony / synchrony (ERD/ERS)
Sox2 (sex-determining region Y-Box) is one of the master transcriptional factors that are important in maintaining the pluripotency of embryonic stem cells (ESCs). In line with this function, Sox2 expression is largely restricted to ESCs and somatic stem cells. We report that Sox2 is expressed in cell lines and tumor samples derived from ALK-positive anaplastic large cell lymphoma (ALK+ALCL), for which the normal cellular counterpart is believed to be mature T-cells. The expression of Sox2 in ALK+ALCL can be attributed to nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), the oncogenic fusion protein carrying a central pathogenetic role in these tumors. By confocal microscopy, Sox2 protein was detectable in virtually all cells in ALK+ALCL cell lines. However, the transcriptional activity of Sox2, as assessed using a Sox2-responsive reporter construct, was detectable only in a small proportion of cells. Importantly, downregulation of Sox2 using short interfering RNA in isolated Sox2active cells, but not Sox2inactive cells, resulted in a significant decrease in cell growth, invasiveness and tumorigenicity. To conclude, ALK+ALCL represents the first example of a hematologic malignancy that aberrantly expresses Sox2, which represents a novel mechanism by which NPM-ALK mediates tumorigenesis. We also found that the transcriptional activity and oncogenic effects of Sox2 can be heterogeneous in cancer cells.
Sox2; transcriptional activity; NPM-ALK; STAT3; tumorigenicity
We report a histologically verified case of ameloblastic fibro-odontosarcoma (AFOS) affecting the right mandible. CT scanning showed an expansile intraosseous mass with ill-defined borders and slightly dense opacities.
jaw; ameloblastic, fibro-odontosarcoma; panoramic radiograph; computed tomography imaging
The purpose of this study was to compare the diagnostic accuracy of cone beam CT (CBCT) with that of conventional dental radiography in the detection of root fractures and to evaluate the influence of root canal fillings on root fracture detection.
We investigated 128 patients with clinically suspected root fractures in 135 teeth. These patients underwent conventional dental radiography, CBCT and eventually surgical exploration. Among the 135 teeth, 86 were non-endodontically treated teeth and 49 were endodontically treated teeth. Two oral radiologists independently analysed the dental radiographs and CBCT images of each patient and reached a consensus. The CBCT findings of root fractures were set as the detection of a separation of the adjacent root segments on at least two contiguous sections and on at least two of the three-dimensional (3D) planes.
Root fracture was intraoperatively detected in 95 of the 135 teeth. The sensitivity and specificity of root fractures diagnosed on the basis of the consensus between the 2 evaluators were 26.3% and 100%, respectively, for dental radiography and 89.5% and 97.5%, respectively, for CBCT. CBCT was significantly more accurate than dental radiography in detecting root fractures (P < 0.001). The sensitivity of CBCT was reduced in the presence of root canal fillings but its specificity remained unaffected. Both the sensitivity and specificity of dental radiography were not influenced by the presence of root canal fillings.
CBCT appears to be more accurate than conventional dental radiography in the detection of root fractures.
cone-beam computed tomography; dental radiography; diagnosis; dental root fracture
We report the case of a woman who had secondary chondrosarcoma in an osteochondroma in the left mandibular condyle, presenting as a large lobulated bone density mass in the left condyle process, to illustrate the clinical characteristics and imaging findings.
secondary chondrosarcoma; mandibular condyle; imaging
To determine the supporting role of a novel foldable capsular vitreous body (FCVB) with magnetic resonance imaging (MRI) in the treatment of severe retinal detachment in human eyes.
The study examined nine eyes of nine patients. Among the nine eyes, five had suffered penetrating injuries while four had suffered contusions of the eyeball involving large defects of the retina or choroids. A standard three-port pars plana vitrectomy was performed, FCVB was triple-folded and sent into the vitreous cavity; balanced salt solution (BSS) was injected into the capsule to support the retina. Three cardinal axes of nine eyes were examined using MRI at baseline and at the 3-month follow up.
MRI revealed that the signal intensity of the FCVB was similar to the normal vitreous body, with low-signal intensity on T1-weighted image and high-signal intensity on T2-weighted image. In three pre-operative silicone oil- or heavy silicone oil-filled eyes, FCVBs were not fully inflated, and eyeball deformation was observed in one eye. Shifts of three cardinal axes of three eyes (horizontal, anteroposterior, and vertical) according to MRI, were −4.33, −4.67, and −2.67 mm. In the remaining six eyes, FCVBs were well distributed in the vitreous cavity and evenly supported the retina; the cardinal axes of the eyes were similar to pre-operation. Shifts of three cardinal axes of six eyes were −0.34, −0.34, and −0.34 mm. In a total of nine eyes, shifts of three cardinal axes were −1.67, −1.77, and −1.11 mm. Statistically significant difference showed only between the horizontal axis of nine eyes pre-operatively and post-operatively (P1=0.041, P2=0.058, P3=0.123).
This study demonstrated the effectiveness of MRI to monitor the supporting role of an FCVB in the treatment of severe retinal detachment in human eyes.
FCVB; human eyes; magnetic resonance imaging
In eukaryotes, mRNA is actively exported to the cytoplasm by a family of nuclear RNA export factors (NXF). Four Nxf genes have been identified in the mouse: Nxf1, Nxf2, Nxf3, and Nxf7. Inactivation of Nxf2, a germ cell-specific gene, causes defects in spermatogenesis. Here we report that Nxf3 is expressed exclusively in Sertoli cells of the postnatal testis, in a developmentally regulated manner. Expression of Nxf3 coincides with the cessation of Sertoli cell proliferation and the beginning of their differentiation. Continued expression of Nxf3 in mature Sertoli cells of the adult is spermatogenesis stage-independent. Nxf3 is not essential for spermatogenesis, however, suggesting functional redundancy among Nxf family members. With its unique expression pattern in the testis, the promoter of Nxf3 can be used to drive postnatal Sertoli cell-specific expression of other proteins such as Cre recombinase.
Sertoli cells; testis; spermatogenesis; Nxf3
During spermiogenesis, haploid round spermatids undergo dramatic cell differentiation and morphogenesis to give rise to mature spermatozoa for fertilization, including nuclear elongation, chromatin remodeling, acrosome formation, and development of flagella. The molecular mechanisms underlining these fundamental processes remain poorly understood. Here, we report that MNS1, a coiled-coil protein of unknown function, is essential for spermiogenesis. We find that MNS1 is expressed in the germ cells in the testes and localizes to sperm flagella in a detergent-resistant manner, indicating that it is an integral component of flagella. MNS1–deficient males are sterile, as they exhibit a sharp reduction in sperm production and the remnant sperm are immotile with abnormal short tails. In MNS1–deficient sperm flagella, the characteristic arrangement of “9+2” microtubules and outer dense fibers are completely disrupted. In addition, MNS1–deficient mice display situs inversus and hydrocephalus. MNS1–deficient tracheal motile cilia lack some outer dynein arms in the axoneme. Moreover, MNS1 monomers interact with each other and are able to form polymers in cultured somatic cells. These results demonstrate that MNS1 is essential for spermiogenesis, the assembly of sperm flagella, and motile ciliary functions.
Cilia are microtubule-based structures present in virtually all cells in vertebrates. Cilia have diverse functions in development, growth, signaling, and fertilization. Primary ciliary dyskinesia (PCD) affects one in 16,000 individuals. PCD is characterized by bronchiectasis and chronic sinusitis, and is often associated with situs inversus and male infertility. The genetic cause of PCD is heterogeneous. Some cases of PCD in humans and animals are caused by single genic mutations such as mutations in genes encoding microtubule-based dynein arm components. We have characterized a protein called MNS1 and found that it plays an essential role in ciliary functions in mice. MNS1 is a novel and integral component of sperm flagella. Mice lacking MNS1 exhibit male sterility as evidenced by abnormal assembly of sperm flagella. MNS1–deficient mice also display defects in left–right asymmetry patterning of internal organs and hydrocephalus. Therefore, mutations in MNS1 may contribute to male infertility and PCD in humans.
Activation of muscarinic acetylcholine receptors is primarily responsible for urinary bladder emptying. Because multiple subtypes of muscarinic receptors exist, we wished to characterize those present in bladder and ultimately to attribute function to those that regulate bladder contractility, neurotransmitter release and perhaps other cholinergic functions in this tissue. Although the m2 and m3 subtypes could be immunoprecipitated after solubilization from human, rat, rabbit and guinea pig bladder membranes, the m1, m4 and m5 subtypes could not. The m2:m3 ratio was 9:1 in rat bladder but was only 3:1 in the other species examined. Immunoprecipitation of the m2 subtype correlated with the relative levels of high-affinity agonist binding sites measured by competition of carbachol for [3H]N-methylscopolamine binding or measured directly using [3H]oxotremorine-M. In the presence of agonist, but not antagonist, GTP binding proteins could be immunoprecipitated in concert with the m2 or m3 receptors using anti-receptor antibodies. These proteins were members of the Gi and Gq/11 subfamilies for both the m2 and the m3 receptor subtypes. In spite of the preponderance of the m2 receptor in all species studied, Schild analysis using somewhat selective antagonists showed that the pharmacologically defined m3 receptor mediated contractility in strips of rat and rabbit bladder. Thus acetylcholine activates bladder smooth muscle via the m3 receptor subtype, and subsequent contractility may be transduced by guanine nucleotide binding proteins such as the Gi and Gq/11 subfamilies.
The aim of this study was to investigate the movement, and the factors that influence such movement, of pancreatic lesions and to provide a reference for determination of planning target volume (PTV) during stereotactic radiotherapy. We implanted 19 gold markers into the inner pancreatic tumours of 16 pancreatic carcinoma patients percutaneously under B-ultrasonographic guidance. The marked motion of pancreatic lesions in the x (right–left), y (superoinferior) and z (anteroposterior) directions was measured using an X-ray simulator system. Based on the statistical analysis of the detected movements, we investigated the relevant influencing factors of pancreatic lesions with multinomial linear regression. Data showed that the mean motion amplitudes of pancreatic lesions were 0.16 cm ± 0.06 (range 0.1–0.3 cm) in the x direction, 0.25 cm ± 0.12 (range 0.1–0.4 cm) in the y direction and 0.88 cm ± 0.24 (0.5–1.6 cm) in the z direction. Motion amplitude was not correlated with the height, weight or age of the patients nor with the location or size of the tumour. The motion of pancreatic lesions was mainly influenced by the respiratory motion and has maximal amplitude in the z direction. Therefore, motion in the z direction should be given a priority consideration while determining the PTV.
To assess the contribution of work-organisational and personal factors to the prevalence of work-related musculoskeletal disorders (WMSDs) among garment workers in Los Angeles.
This is a cross-sectional study of self-reported musculoskeletal symptoms among 520 sewing machine operators from 13 garment industry sewing shops. Detailed information on work-organisational factors, personal factors, and musculoskeletal symptoms were obtained in face-to-face interviews. The outcome of interest, upper body WMSD, was defined as a worker experiencing moderate or severe musculoskeletal pain. Unconditional logistic regression models were adopted to assess the association between both work-organisational factors and personal factors and the prevalence of musculoskeletal pain.
The prevalence of moderate or severe musculoskeletal pain in the neck/shoulder region was 24% and for distal upper extremity it was 16%. Elevated prevalence of upper body pain was associated with age less than 30 years, female gender, Hispanic ethnicity, being single, having a diagnosis of a MSD or a systemic illness, working more than 10 years as a sewing machine operator, using a single sewing machine, work in large shops, higher work–rest ratios, high physical exertion, high physical isometric loads, high job demand, and low job satisfaction.
Work-organisational and personal factors were associated with increased prevalence of moderate or severe upper body musculoskeletal pain among garment workers. Owners of sewing companies may be able to reduce or prevent WMSDs among employees by adopting rotations between different types of workstations thus increasing task variety; by either shortening work periods or increasing rest periods to reduce the work–rest ratio; and by improving the work-organisation to control psychosocial stressors. The findings may guide prevention efforts in the garment sector and have important public health implications for this workforce of largely immigrant labourers.
work organisation; musculoskeletal disorder; ergonomic; psychosocial; sewing machine
To describe the patterns of seat belt wearing in Nanjing, China for drivers, front seat passengers, and rear occupants of motor vehicles.
Roadside observational study.
Four sites in central and northern Nanjing during daylight hours over 1 week in April 2005.
Drivers and passengers of 17 147 cars, taxis, goods vans, and pickups, which traveled in the inside traffic lane.
Main outcome measures
Percentage seat belt wearing for each of seating position, age/sex, time of day, vehicle type, day of week.
The rate of seat belt wearing was significantly higher in drivers (67.3%, 95% CI 66.6 to 68.0) than front seat passengers (18.9%, 95% CI, 18.0 to 19.8). It was negligible for second front seat passengers (2.6%, 95% CI 0.3 to 4.9) and rear seat passengers (0.5%, 95% CI 0.3 to 0.7). Belt tampering, such that protection would be reduced in the event of a crash, was observed for 18.5% of taxi drivers. Drivers were most likely to wear seat belts in cars and vans and at a city roundabout; front seat passengers were most likely to wear seat belts in non‐taxi vehicles, during the evening rush hour, if the driver was wearing a belt, and on the local north road. Drivers were least likely to wear a belt in the early morning, in pickups and taxis, on Tuesday (or the following week), and on the local north road; front seat passengers were least likely to wear a belt in taxis and if the driver was not wearing a belt.
Rates of seat belt wearing by passengers were low despite national legislation and provincial regulations coming into effect several months before the survey. Combined education and enforcement are necessary accompaniments to legislation.
Meiosis is unique to germ cells and essential for reproduction. During the first meiotic division, homologous chromosomes pair, recombine, and form chiasmata. The homologues connect via axial elements and numerous transverse filaments to form the synaptonemal complex. The synaptonemal complex is a critical component for chromosome pairing, segregation, and recombination. We previously identified a novel germ cell–specific HORMA domain encoding gene, Hormad1, a member of the synaptonemal complex and a mammalian counterpart to the yeast meiotic HORMA domain protein Hop1. Hormad1 is essential for mammalian gametogenesis as knockout male and female mice are infertile. Hormad1 deficient (Hormad1−/−) testes exhibit meiotic arrest in the early pachytene stage, and synaptonemal complexes cannot be visualized by electron microscopy. Hormad1 deficiency does not affect localization of other synaptonemal complex proteins, SYCP2 and SYCP3, but disrupts homologous chromosome pairing. Double stranded break formation and early recombination events are disrupted in Hormad1−/− testes and ovaries as shown by the drastic decrease in the γH2AX, DMC1, RAD51, and RPA foci. HORMAD1 co-localizes with γH2AX to the sex body during pachytene. BRCA1, ATR, and γH2AX co-localize to the sex body and participate in meiotic sex chromosome inactivation and transcriptional silencing. Hormad1 deficiency abolishes γH2AX, ATR, and BRCA1 localization to the sex chromosomes and causes transcriptional de-repression on the X chromosome. Unlike testes, Hormad1−/− ovaries have seemingly normal ovarian folliculogenesis after puberty. However, embryos generated from Hormad1−/− oocytes are hyper- and hypodiploid at the 2 cell and 8 cell stage, and they arrest at the blastocyst stage. HORMAD1 is therefore a critical component of the synaptonemal complex that affects synapsis, recombination, and meiotic sex chromosome inactivation and transcriptional silencing.
The biology of germ cells is intimately intertwined with meiosis. Meiosis I is a unique biological event, when chromosomes pair, recombine, and segregate. The synaptonemal complex is a protein lattice that enables chromosome pairing and recombination and is unique to meiosis I. Meiosis I requires a subset of factors that are unique to germ cells and meiosis. Germ cell–specific factors are known to play crucial roles during formation of the synaptonemal complex and include synaptonemal complex proteins SYCP1, SYCP2, and SYCP3, among others. We discovered a mouse HORMA domain containing protein, Hormad1 (Nohma), which is germ cell–specific and essential for male and female fertility. Mice deficient in Hormad1 have severe defects in early recombination, synapsis, and segregation—functions attributed to yeast HORMA domain containing protein, Hop1. Moreover, Hormad1 is likely a germ cell–specific component of the meiotic sex chromosome inactivation and transcriptional silencing complex.
Tumor necrosis factor-α (TNF-α) is a cytokine that has an important role in immunity and inflammation by inducing cellular responses such as apoptosis. The transcription factor nuclear factor-κB (NF-κB) can paradoxically suppress and promote apoptosis in response to TNF-α. In this study, we found that p53 upregulated modulator of apoptosis (PUMA), a p53 downstream target and a BH3-only Bcl-2 family member, is directly regulated by NF-κB in response to TNF-α. TNF-α treatment led to increases in PUMA mRNA and protein levels in human colon cancer cells. The induction of PUMA was p53 independent, and mediated by the p65 component of NF-κB through a κB site in the PUMA promoter. The apoptotic effect of PUMA induction by TNF-α was unmasked by depleting the antiapoptotic protein Bcl-XL. In mice, PUMA was also induced by TNF-α in an NF-κB-dependent manner. TNF-α-induced apoptosis in a variety of tissues and cell types, including small intestinal epithelial cells, hepatocytes, and thymocytes, was markedly reduced in PUMA-deficient mice. Collectively, these results demonstrated that PUMA is a direct target of NF-κB and mediates TNF-α-induced apoptosis in vitro and in vivo.
PUMA; NF-κB; TNF-α; apoptosis