This investigation first set out to analyse which cellular proliferative and apoptotic factors, in addition to the clinical prognostic factors, are most predictive in patients with non-small-cell lung carcinomas (NSCLC). To this purpose, we related the proliferative factors proliferating cell nuclear antigen (PCNA), cyclin A, cyclin D1, cyclin-dependent kinase 2 (cdk2), cdk4 and the proportion of cell cycle phases in NSCLC to the survival times of 150 patients. Additionally, we associated the expressions of Fas, Fas ligand and caspase-3 in NSCLC to patient survival. Immunohistochemistry was used to determine the proteins and flow cytometry to assess the proportion of cell cycle phases. Patients with PCNA-positive carcinomas had significantly shorter survival times than patients with PCNA-negative carcinomas . Corresponding results were obtained with the factor cyclin A (64 vs 92 weeks), with the factor cdk2 (76 vs 89 weeks), with the factor cdk4 (62 vs 102 weeks) and with the proportion of S phases (86 vs 121 weeks). Patients with an expression of the apoptotic factors had a more favourable prognosis than patients with negative carcinomas. The median survival times of cancer patients with Fas expression was 86 weeks and of those without Fas expression only 69 weeks. Corresponding results were obtained with the Fas ligand (87 vs 41 weeks) and caspase 3 (87 vs 34 weeks). In order to determine whether a combination of factors can yield improved prognostic information, we investigated all possible combinations of the proliferative and apoptotic factors. Patients with tumours having a high proliferative activity, but which did not express apoptotic factors had the shortest survival times while patients with a low proliferative activity and a high expression of apoptotic factors had the most favourable outcome. A multivariate analysis (Cox model) of the cellular and clinical prognostic factors indicated that stage, lymph node involvement, Fas, PCNA and cyclin A are the most important prognostic factors for the clinical outcome of patients with non-small-cell lung carcinomas. © 2000 Cancer Research Campaign


Epidemiological evidence has implicated Chinese salted fish as a human nasopharyngeal carcinogen. In the present study, 221 Wistar-Kyoto rats aged 21 days were randomly assigned to one of three experimental groups. Rats in group 1 (high dose group) were fed a powder diet of one part Chinese salted fish to three parts certified rat chow during the first 18 months. Similarly, rats in group 2 (low dose group) were fed a powder diet of one part salted fish to five parts rat chow for 18 months. Rats in group 3 were given rat chow only throughout the 3-year experiment. Four malignant tumours of the nasal cavity were observed among rats fed the experimental diets (three and one respectively in the high and low dose groups). No comparable tumours were observed in controls, compatible with the historical control rate of zero. Our results, therefore, further strengthen the hypothesis that Chinese salted fish is a human nasopharyngeal carcinogen; they also establish Wistar rats as a viable animal model for carcinogenicity studies of this food in the laboratory.


From 1996 through 1999, 35 deer mice (Peromyscus maniculatus) were captured in 25 urban and suburban homes in southwestern Montana. Mice were captured throughout the year except for January; seven mice (20%) from seven (28%) of the homes were seropositive for Sin Nombre virus. The infected mice were mostly adult males captured in the spring and fall.


A new autofluorescence (AF) system for bronchoscopy that operates as compact as aconventional white light bronchoscopy system is described. The system is also capable of whitelight illumination and excitation of aminolevulinic acid (ALA) induced fluorescence. Changing between white light and (auto-) fluorescence mode is easy and always possible. Broad band excitation with blue light  delivers highintensity illumination at the distal end of the bronchoscope . A specialoptical observation technique makes the AF directly visible to the eye instead of indirecttechniques used in other AF systems. A compact (160 g)and sensitive camera can be used for documentation.


No numerical aberration of chromosomes that might be specific for prostate cancer has so far been established. We used fluorescence in situ hybridisation (FISH) with centromere-specific probes for chromosomes 7, 8, 17, X and Y to establish the distribution of centromere copy numbers in frozen-stored or freshly prepared samples of benign prostate hypertrophy (BPH) and to detect numerical aberrations of these chromosomes in 28 prostate cancers from Japanese men. There was no significant difference in the data of centromere copy numbers between fresh and frozen-stored tissue. The most common aberration in prostate cancers was a gain of chromosome 8 (57%), with numerical aberration of chromosome 7 being the second most frequent anomaly (50%). Numerical aberration of chromosome 7 is most significantly associated with a higher Gleason score (GS) (P < 0.005) or with lymph node metastasis (P < 0.001). Numerical aberration of several chromosomes, including chromosomes 7 and/or 8, was common in aggressive prostate cancers. Loss of chromosome Y was detected in only 4% of cases. FISH analysis thus proved to be a useful method for detecting numerical aberrations of individual chromosomes, with application to touch preparations of frozen-stored tissue having the advantage of exact sampling of cancer foci. The results suggest that numerical aberration of chromosome 7 is associated with aggressive tumour behaviour and poor prognosis of patients with prostate cancer. The association between genetic change and chromosomal abnormality should be studied in detail.


Idiopathic juxtafoveolar retinal telangiectasis (IJRT) group 1A usually affects one eye of young and middle aged male patients.1 TelangieA 40-year-old healthy male was referred to us for visual distortion and inability to see in the upper left field of vision in the right eye (OD) for six weeks. Medical work-up and laboratory tests excluded the presence of any systemic ailment including hypertension and diabetes mellitus. The best-corrected visual acuity was 20/30 OD and 20/20 in the left eye (OS). Anterior segment examination showed no cell or flare in the anterior chamber or in the anterior vitreous in both eyes (OU). Intraocular pressure (IOP) was 15 mmHg OU. Dilated fundus examination OD showed abnormal telangiectatic blood vessels and hard exudates temporal to the fovea as well as cystic appearance in the foveal area . The funOver the next one year the patient was followed closely without improvement in vision (following laser treatment) or change in the fundus appearance. However, 15 months after the initial examination, his vision dropped to 20/50 OD with significant increase in macular edema clinically and on OCT . The cenOn the basis of biomicroscopic and fluorescein angiographic patterns, Gass and Blodi have classified IJRT into three groups: Group 1 Laser photocoagulation and photodynamic therapy of the telangiectatic capillaries in IJRT group 1A are reported to be effective in reducing the foveolar edema and exudation.–4 Howeveet al. and Maia et al. successfully treated their patients of group 1 IJRT with intravitreal injection of triamcinolone acetonide.[There was dramatic improvement in vision two weeks following intravitreal triamcinolone injection in the present case. The vision improved along with resolution of macular edema that was confirmed on clinical examination and on OCT. Li cetonide.6 Both thOur experience with this patient demonstrates that intravitreal triamcinolone acetonide may be considered an additional therapeutic option besides photocoagulation in the management of IJRT group 1A, especially when the latter treatment is ineffective. Moreover, OCT is a useful non-invasive tool that helps in diagnosing the disease and following it over the years. Further long-term case controlled prospective studies evaluating the safety and efficacy of this treatment modality for patients with IJRT group 1A are warranted.


In vivo growth characteristics of myeloid leukaemia induced by whole-body irradiation of CBA/H male mice were examined in the strain of origin by procedures expected to enhance or depress immunological responses. Syngeneic growth in vivo  was not modified by attempted active immunization with radiation-inactivated cells or by sublethal whole-body irradiation of recipients before inoculation of small numbers of clonogenic cells. Since the growth stimuli involved in in vivo repair of severely damaged normal haemopoietic tissue also did not modify the growth of the radiation-induced leukaemia cells in syngeneic passage, their growth in vivo in the irradiated primary hosts can be regarded as autonomous by the stage at which leukaemia was diagnosed. Challenge inocula in the "immunization" experiments were 1-9 clonogenic cells from 4 different passaged lines and in the whole-body radiation experiments, 1-10(3) clonogenic cells derived from 11 different primary hosts and 4 different passaged lines.


Introduction and aim. Management of complicated wounds after tumor extipiration of pelvic and proximal lower limb musculoskeletal sarcoma represents an essential component in the outcome of these patients. The authors present modified vertical rectus abdominis musculocutaneous (VRAM) flap techniques to reconstruct extensive defects after debridment of these complicated wounds. Material and Methods. Over a period of 4 years (2002–2005), 5 men and 2 women were managed. Median age was 21 years (range 15–49). The patients were managed for complicated lower trunk, groin, and upper thigh wounds after resection of three pelvic chondrosarcomas as well as two pelvic and two proximal femur osteosarcomas. The modifications included a VRAM flap with lateral and tongue-like extension design of the skin paddle (5 cases) or a delayed extended VRAM flap (2 cases). Results. All flaps showed complete survival and healing with no ischemic events providing stable coverage. All patients were ambulant with good limb functions in terms of walking and gait after adequate rehabilitation, 2 needed support with crutches. Conclusion. The modified VRAM flaps offer reliable reconstructive tools for coverage of complex groin and thigh defects by providing larger well-vascularized soft tissue with acceptable donor site. Limb salvage remains a major objective in contemporary management ofbony and soft tissue sarcomas of the pelvis and lower extremities , 2. RadiSeven consecutive patients with complicated wounds secondary to resection of pelvicand proximal lower extremity musculoskeletal sarcoma were managed with modified VRAM flaps between2002–2005. The medianage of the patients in this series at the time of surgery was 21 years (range15–49) and included 5 men and 2 women. The modifications included VRAM flapwith lateral and tongue-like extension design of the skin paddle (5 cases) or adelayed extended VRAM flap (2 cases). All the patients have been treated with amultidisciplinary approach; the management of osteosarcoma patients  included a 4-month course of neoadjavant chemotherapyfollowed by resection of the primary tumor and postoperative 3-month course of chemotherapy;while chondrosarcoma patients (3 pelvic) were offered surgical resection only.The complications, which necessitated management by the plastic andreconstructive surgeons, included infection and wound dehiscence with ultimateexposure of metal prosthesis (4 patients) and necrosis of thigh flaps (3patients). The median interval duration between the primary operation and thereconstructive procedure for the complicated wounds was 16 days (range: 13–36).All patients underwent thorough clinical assessment, adequate debridment andirrigation of the wounds, followed by immediate coverage of the resultantdefects (median size 7 × 17 cm) usingmodified VRAM flaps.This technique was used to cover exposed metal plates dueto wound breakdown after osteosarcomas resection of proximal femur where lengthof the flap was predominantly required than its width. An ipsilateralparasternal skin paddle was designed 10 cm cephalic to the costal margin and 2 cm from the lateral sternal edge. The pinch test, originally described byTaylor et al. , was useThis technique was used in cases after resection ofchondrosarcomas and osteosarcomas of the pelvis, which necessitated thedevelopment of large subcutaneous thigh flap to allow adequate exposure andvascular exploration to achieve adequate oncological resection. Extensive full-thicknessthigh flap necrosis occurred in these patients which necessitated adequatedebridement and coverage of the resultant defect utilizing a contralateral,laterally oriented VRAM with a tongue-like skin extension. The contralateralmuscle was selected due to the sacrifice of the ipsilateral inferior epigastricpedicle as part of the radicality of the primary tumor resection. The lateralorientation of the skin paddle was designed to be oblique (45°) tothe longitudinal axis of the rectus muscle to provide an average extra lengthof 5 cm. The tongue-like extension ofthe skin paddle was designed throughout the whole length of the muscle taperingtowards the pivot point of the flap at the entry of the inferior epigastricpedicle. The fasciomusculocutaneous component of the VRAM flap was harvestedutilizing the standard technique with elevation of the laterally oriented partof the flap in the fascial plane. Theorigin and insertion of rectus muscle were transacted leaving the vascular pedicle asthe sole tether point; this significantly increased the arc of rotationallowing more muscle mass to be transposed to the wound. Incising the skinbridge between the donor and recipient defect to lay open the tunnel connectingthe recipient and the donor site followed this. The tongue-like skin extension provided the roof foropened tunnel after rotation of the flap to settle to the defect; allowing moreroom for the pedicle and providing coverage for the medial part of the defect.In all patients, the donor site was closed primarilyafter reinforcement of the abdominal wall using an onlay prolene mesh, suctiondrains were used both at the recipient and donor sites. All patients weremobilized within 2 weeks postoperatively under supervision of the orthopaedicand physical therapy team. The mean duration to postoperative chemotherapy was3 weeks. The mean follow-up period was 26 months at which patients were followed up with assessment of the oncological outcome, durability of soft tissue reconstruction, and finally assessing the functional outcome using Enneking system,All flaps showed complete survival and healing withno ischemic events, hence providing stable coverage. Mild infection wasobserved in 1 patient; no haematomas were detected. All patients were ambulantwith good limb functions in terms of walking and gait after adequaterehabilitation; 2 needed additional support with crutches. All patients showedgood emotional acceptance; and no pain has been observed during ambulation inall patients. One patient developed incisional hernia at the donor site that requiredrepair 1 year later; otherwise, no other donor site morbidity was noted.Another patient died of haematogenous metastatic spread after completeresection 15 months later. There was no hypertrophic scar formation or flapbreakdown experienced on long-term follow-up in any of the patients.Limb preservation has become a more realistic andnecessary goal in the management of patients with musculoskeletal sarcoma ofextremities . RadiatiThe reconstructive options to provide coveragefollowing debridment of these complicated wounds would be local, distant flaps,or free-tissue transfer. VRAM flaps havebeen used successfully in coverage of defects of the chest wall, groin, hip,perineal, vaginal, and gluteal regions with good functional outcomes –17. The The delayed phenomenon has been used for many yearsto enhance functional blood flow through vasodilatation of the arterial networkand avoidance of complete vasoconstriction caused by catecholamine release, andhence it increases the flap reliability especially in high-risk patients , 24. TheThe authors believe that these modifications willprovide reliable extralength of soft tissue, which is tension free with a widerarc of rotation, better filling of dead space with well-vascularized tissue soas to resist infection, and stable coverage with good functional outcome.Other modalities described for management ofcomplicated infected wounds as skin grafting or vacuum-assisted closure wouldnot be suitable due to the presence of dead space and exposed hardwareprosthesis after debridement , 4, 27. The use of free-tissue transfer is rare in theseclinical scenarios; on the other, hand-free osteocutaneous fibular flaps wereperformed for primary skeletal reconstruction after tumor extipiration ofsarcoma of the lower extremities  rather tIn conclusion, the modified VRAM flaps would offerreliable reconstructive tools for coverage of complicated primary wound withnecrosis and breakdown after radical resection of pelvic and proximal lowerlimb musculoskeletal sarcoma. They offer larger well-vascularized soft tissuewith acceptable donor site, more durable coverage with no delay in thepostoperative adjuvant therapy, good functional outcome, together with overallimprovement in the survival.


A method was developed for delineating the structure ofparamagnetic metal complexes. The selective disappearanceof cross-peaks in proton-carbon shift correlated 2D NMRmaps was shown to uniquely depend upon the scalar and/ordipolar interaction between ligand nuclei and the unpairedelectron(s), thus providing a means of identifying bindingsites. Copper(II) was shown to form metal complexes withboth Penicillin (PNC) and Pilocarpine (PLC) and the structureof the two 1:2 complexes in water solution at physiologicalpH were determined.


Solid tumours develop an acidic extracellular environment with high concentration of lactic acid, and lactic acid produced by glycolysis has been assumed to be the major cause of tumour acidity. Experiments using lactate dehydrogenase (LDH)-deficient ras-transfected Chinese hamster ovarian cells have been undertaken to address directly the hypothesis that lactic acid production is responsible for tumour acidification. The variant cells produce negligible quantities of lactic acid and consume minimal amounts of glucose compared with parental cells. Lactate-producing parental cells acidified lightly-buffered medium but variant cells did not. Tumours derived from parental and variant cells implanted into nude mice were found to have mean values of extracellular pH (pHe) of 7.03 +/- 0.03 and 7.03 +/- 0.05, respectively, both of which were significantly lower than that of normal muscle . Lactic acid concentration in variant tumours (450 +/- 90 microg g(-1) wet weight) was much lower than that in parental tumours (1880 +/- 140 microg/g(-1)) and similar to that in serum (400 +/- 35 microg/g(-1)). These data show discordance between mean levels of pHe and lactate content in tumours; the results support those of Newell et al (1993) and suggest that the production of lactic acid via glycolysis causes acidification of culture medium, but is not the only mechanism, and is probably not the major mechanism responsible for the development of an acidic environment within solid tumours.


Automated Flow Injection Gas Diffusion (FIGD) promotes theselective transmembrane diffusion of MAs and NH3 from aqueoussample under strongly alkaline , chelated (EDTA) conditions into a recycled acidic acceptor stream. The acceptor is then injected onto an ion chromatograph where NH3 and the MAs are fully resolved as their cations and detectedconductimetrically. A versatile PC interfaced control unit and data capture unit (DCU) are employed in series to direct the selonoid valve switching sequence, IC operation and collection of data. Automation, together with other modifications improved bothlinearily  andprecision (<8%) of FIGD-IC at nanomolar concentrations, compared with the manual procedure. The system was successfully applied to the determination of MAs and NH3 in seawater and in trapped particulate and gaseous atmospheric samples during anoceanographic research cruise.The automation and improved design and performance of FlowInjection Gas Diffusion-Ion Chromatography (FIGD-IC), a novel technique for the simultaneous analysis of trace ammonia (NH


A simple, rapid histone agglutination test (HAT) is described. It consists of incubation in microplates or in microtubes of blood lymphocytes isolated from cancer patients and patients with non-malignant diseases with microquantities of histone fraction F2A1. Positive reaction is shown by massive agglutination of lymphocytes of the patients tested (126 subjects): this test was positive in 76% of cases. All controls (59 subjects) were negative.


By analogy with the case of [Pt(en)2]Cl2 it is suggested that Auen acts as a minor groove binder.Reactions of the complexes bisethylendiammine gold(III) and bisethylendiammine platinum(II) with calfthymus DNA were comparatively analysed. Both complexes bind DNA non-covalently most probably on the basis of electrostatic interactions. Binding of either complex at low ratios results into modest modifications of B-type DNA conformations, as detected by CD. Far larger CD alterations are observed at high ratios. The gold(III) chromophore is scarcely perturbed by DNA addition Binding of [Au(en)


During a 15-month period, we used in vivo bromodeoxyuridine (BUDR) infusion to study cell kinetics in 112 consecutive patients with various types of malignant tumours: acute leukaemia (50 patients), gastric cancer (42) and brain gliomas (20). The in vivo BUDR method requires that a single tumour sample be taken 4-6 h after infusion and that bivariate flow cytometry (FCM) be employed to measure simultaneously the percentage of BUDR-labelled cells  and their mean DNA content (following propidium iodide staining). This technique rapidly furnishes the labelling index (LI) and the DNA synthesis time (TS), from which the tumour potential doubling time (Tpot) and production rate  are calculated. The procedure took 6-9 h to complete and there was no immediate toxicity from BUDR administration. Successful LI and TS determinations were obtained in 89 (80%) and 80 (72%) of the 112 patients, respectively. Correlations were sought between kinetic parameters and a number of pathological and clinical ones. In 34 patients with acute non-lymphoblastic leukaemias who were uniformly treated for remission (CR) induction and maintenance, proliferative activity, as measured by Tpot and FTR, was greater in responsive than in non-responsive patients, and in those who experienced CR for over 8 months than in those who had a shorter CR. Proliferative activity was also greater in patients with advanced gastric cancers than in those with more limited disease. No correlations between kinetic and clinical and pathological parameters were found in gliomas. These data indicate the in vivo BUDR infusion coupled with FCM measurements can be performed in clinical settings to obtain kinetic data rapidly in quite large patient series. This will probably allow the inclusion of kinetic data in clinical trials aimed at evaluating the prognostic relevance of these data.


Long-term cognitive and educational sequelae have been inconsistently reported in children who received cranial irradiation (CRT) to prevent central nervous system (CNS) disease in acute lymphoblastic leukaemia (ALL). This study investigates a large and representative sample of survivors of ALL and compares them with non-irradiated survivors of cancer and healthy control children to determine the effect of CRT on cognitive and educational ability. Three groups of children were studied: Group 1 (n=100) survivors of ALL treated with chemotherapy and CRT, group 2 (n=50) children with a variety of malignancies treated with chemotherapy alone, group 3(n=100) healthy children. Cognitive and educational abilities of these groups were evaluated using standardised psychometric techniques. Significant differences in cognitive and educational abilities were found between the children in group 1 (chemotherapy + CRT) and the two control groups, with the children receiving CRT performing less well in a range of tests. Greatest differences were detected for tasks dependent on language function including verbal IQ, reading and spelling. Within group 1 a younger age at treatment (less than 5 years) and a higher dose of CRT (24 Gy vs 18 Gy) were predictive of poor long-term outcome for cognitive and education ability. In contrast, children who received chemotherapy alone, with or without intrathecal methotrexate, performed similarly to healthy controls. No gender differences were detected for these measures.


Alterations in DNA ploidy accompany hepatocellular carcinoma (HCC). However, changes in DNA content are also seen in regenerating liver and with increasing age. Thus, to investigate the role of DNA ploidy changes in development of HCC, flow cytometric DNA content determinations were done in a rat model system of peroxisome proliferator-induced HCC. Paraffin blocks of liver isolated from 18 Fisher 344 male rats fed ciprofibrate for 20 weeks (4), 40 weeks (4) or 20 months (10) were examined. Livers from age-matched control rats were also examined. From the 20 month ciprofibrate group, nine neoplastic nodules (NNs), 27 HCCs and four non-tumorous surrounding tissue controls (NTCs) were examined. Significant DNA tetraploid populations were seen in both the NNs and NTCs. A significant increase in the percentage of DNA diploid cells was observed in the NN samples. No significant difference in the percentage S-phase cells was seen. Emergence of cell populations with new DNA ploidy classes (8c or DNA aneuploid) as compared with NTCs was only seen in HCCs (7 of 27), and five of these seven were DNA aneuploid, as distinct from DNA tetraploid, populations. A total of 16 of 24 HCC samples that were adequate for cell cycle analysis had average percent S-phase greater than the mean of the NTCs plus three standard deviations. Although a direct role cannot be inferred, these results support the hypothesis that increases in the fraction of diploid cells is an important early event in the development of rat HCC and that further alterations in DNA ploidy and increased proliferative fraction accompany the development of HCC.


Copper(II) complexes of several hydroxyflavones were prepared and characterised through their physico-chemical properties. The nuclease activity of three synthesised complexes is reported. These copper(II) complexes present more nuclease activity than the ligands and the copper(II) ion.


We examined hepatic artery ligation combined with an angiogenesis inhibitor, TNP-470, in the treatment of VX2 tumour inoculated into the liver of rabbits. Effects on tumour growth were correlated with arterial collateral development in this system. Three treatment methods were compared: (1) the left hepatic artery was ligated at the liver hilum (ligation group); (2) TNP-470 (40 mg per body) was infused continuously for 7 days via the common hepatic artery (TNP group); (3) the left hepatic artery was ligated and TNP-470 was infused continuously for 7 days via the common hepatic artery (ligation + TNP group). These treatments were started 12-14 days after tumour inoculation. The day of initiating treatment was defined as day 0. Although there were no significant differences in tumour volume among the three treated groups on day 7 after treatment, tumour volumes in the ligation + TNP group were significantly smaller than in the ligation group and the TNP group on day 14 after treatment. The vasculature and arterial collaterals around the tumour were demonstrated by the perfusion of a silicon rubber solution, Microfil. In the ligation + TNP group, the new microvasculature around the tumour decreased compared with the ligation group. The TNP-470 inhibition of microvascular proliferation may limit the development of collaterals that communicate with new feeding arteries. These results suggest that transarterial embolization combined with TNP-470 may enhance the anti-tumour effect of transarterial embolization alone in the treatment of liver tumours.


The hormonal mechanisms involved in the development of gynaecomastia accompanying the treatment of multiple myeloma in adult men have been investigated by studying levels of circulating testosterone (T), oestrone (EI), oestradiol (E2), sex-hormone binding globulin (SHBG), prolactin (PRL) and the gonadotrophins LH and FSH, before, during and after development of gynaecomastia in 4 men. These have been compared with 5 closely matched men who did not develop gynaecomastia during similar treatment for myeloma. Levels of circulating T fell, and levels of E1 and E2 rose during treatment periods in all subjects, and the changes were statistically significant in subjects developing gynaecomastia, which resolved as levels of sex steroid returned towards normal following cessation of treatment. We conclude that treatment of adult men for myeloma results in testicular dysfunction with a reduction in circulating T and a rise in circulating oestrogens. These changes are most marked in subjects developing gynaecomastia in whom the normal breast tissue is stimulated by a subtle, transient oestrogen:androgen imbalance in favour of oestrogens.


Twenty-four symptomatic patients with advanced non-small cell lung cancer (NSCLC) were treated with cisplatin-based chemotherapy . Patients were assessed for symptom relief as well as for objective response. Although only five patients achieved an objective response (21%), 18 patients (75%) reported a complete disappearance or good improvement in at least one of their tumour-related symptoms. The overall symptomatic response rate was 67% with 16 patients feeling better or much better on treatment. The toxicity of treatment (primarily myelosuppression and nausea and vomiting) was mild and hair loss was minimal. The high incidence of symptomatic relief seen in this study, even in the absence of objective response, suggests that moderate dose chemotherapy may have a role in the palliation of NSCLC.


Activation of the RET tyrosine kinase domain occurs in a proportion of thyroid papillary carcinomas. Three chromosomal rearrangements have been described, of which PTC1 is the commonest. Wide differences (2.5-25%) in frequency of PTC1 in different populations have been reported; it is not clear whether these are due to environmental factors, racial differences or technical reasons. We have developed a simple and rapid reverse transcriptase nested polymerase chain reaction (RT-nPCR) method enabling the detection of gene expression from single 5 microns sections of formalin-fixed paraffin wax-embedded archival material. We have applied this approach to detect expression of the RET tyrosine kinase domain, allowing identification of RET activation resulting from any rearrangement, whether characterised or not, or from overexpression. A retrospective study was performed on 22 adult and 21 childhood papillary carcinomas. Thirteen of 22 (59%) adult and 10 of 21 (48%) childhood carcinomas showed evidence of RET activation, demonstrating a major role for the RET oncogene in UK thyroid papillary carcinogenesis. This study also shows a similar frequency of RET activation in both children and adults. The use of a technique that allows reliable amplification of RNA from archival material, using primers chosen in different exons so that amplified products are readily distinguished from genomic DNA, will allow correlation of translocations and chromosomal rearrangements with a variety of specific tumour types.


A lymphocyte stimulation assay is described which detects immune reactivity to antigens derived from the CaSki cervical carcinoma cell line. Taking a stimulation index of greater than 4.1 as positive, the peripheral blood lymphocytes of 14/20 patients (70%) with untreated dysplasia or carcinoma-in-situ, 8/19 patients (42%) with untreated macroinvasive squamous cell carcinoma of the uterine cervix and 8/38 controls (21%) showed positive reactions. Statistical analysis revealed a significant difference between the group of patients with dysplasia or carcinoma-in-situ and the controls. The sera of patients and controls were simultaneously tested for the presence of tumour-directed antibody. There was no correlation between the occurrence of cellular immune reactivity and of serum antibody, both directed to cervical carcinoma antigens. Cellular immune reactivity tended to occur more frequently in patients with preinvasive stages of cervical carcinoma, and serum antibody in patients with macroinvasive carcinoma.


These data stimulate further epidemiological and functional studies focusing on the role of homocysteine and folate metabolism in lymphoma tumorigenesis.Primary central nervous system lymphomas (PCNSL) frequently reveal genomic instability. We analysed different functional genetic variants affecting the folate and homocysteine metabolism important for DNA integrity in 31 PCNSL patients and 142 controls. We found significantly less carriers of the methionine synthase c.2756A>G (D919G) missense polymorphism among the patients (0.16  S-adenosylmethionine (SAM) as a cosubstrate in DNA methylation.Primary central nervous system lymphomas (PCNSL) are highly malignant non-Hodgkin's lymphomas (NHL). The vast majority of them are of the diffuse large B-cell type . GeneticFigure 1We performed a case–control study using DNA samples of 31 German patients with PCNSL of the DLBCL-subtype .The Hardy–Weinberg equilibrium was confirmed for all polymorphisms. Allele frequencies of the control group did not significantly differ from those reported in the literature  or χ2-analysis . Further, we did not find any significant association of the other polymorphisms  or combinations with PCNSL.The CBS 844ins68 polymorphism was detected more frequently in patients (26%) than in controls (14%). However, the difference did not reach statistical significance in regression analysis  is associated with a lower risk of developing colorectal cancer and types of systemic NHL  against PCNSL and encourage future studies investigating the influence of genetic and dietary conditions of homocysteine and folate metabolism on cancerogenesis.


Serologic and molecular evidence indicates that peste des petits ruminants virus (PPRV) infection has emerged in goats and sheep in the Ngari region of southwestern Tibet, People’s Republic of China. Phylogenetic analysis confirms that the PPRV strain from Tibet is classified as lineage 4 and is closely related to viruses currently circulating in neighboring countries of southern Asia. Peste des petits ruminants (PPR) is an acute and highly contagious viral disease contracted by small ruminants such as goats and sheep and causing high rates of illness and death. The disease is endemic in parts of sub-Saharan Africa, the Middle East, and Asia. The PPR virus (PPRV) genogroup consists of 4 lineages  in the Ngari region, where animals having clinical signs of PPRV infection had been reported by local authorities. The second group included 298 animals in Gar and Bulang counties in the same region and in 2 counties bordering the Ngari region (Nyima in Nagqu region and Zhongba in Shigatse region). The third group contained 520 animals in 5 counties within 3 separate regions . We examined 1,536 animals (771 goats and 765 sheep) and collected serum samples from each. A competitive ELISA that used a monoclonal antibody to the N protein . Rates oField samples, including organ  and swab specimens, were obtained from 49 goats and sheep suspected of being infected with PPRV. These animals inhabited 4 counties in the Ngari region . Two reverse transcription–PCRs (RT-PCR) and 1 newly developed and validated real-time quantitative RT-PCR (qRT-PCR) were conducted to determine whether the animals had viral RNA .The study confirmed 11 outbreaks in 4 counties in the southwest Ngari region in Tibet. Nine of the 11 occurred in southern Rutog County, northern Ge’gyai County, and Gerze County, and samples from these 9 were seropositive for PPRV , panel AThe nucleic acid sequences obtained from the PCR products were aligned with sequences from PPRV strains available in GenBank. Partial sequencing (448 bp) of the F gene showed that 20 of 21 samples were identical over the portion of the genome that was characterized (GenBank accession no. EU816772). One (GenBank accession no. EU815053) differed from other Ngari sequences by 1 nt. The Ngari sequences showed a level of nucleotide identity with other PPRV strains of 88.8%–98.8%. Strains of PPRV from Tibet were classified as lineage 4 and were closely related to the India/Bsk/Guj/05 strain isolated in India in 2005 , panel AIn this study, PPRV was found by collecting samples from animals in the field and detecting infection by using competitive ELISA and RT-PCR. Our research provides valuable data on PPRV infection in small ruminants in Tibet. Infection was observed in 4 counties in the Ngari region of southwestern Tibet. Most outbreaks occurred in Rutog and Ge’gyai counties; 1 outbreak was confirmed in Gerze County and another in Zada County. Epidemiologic and serologic evidence suggests that the infection first emerged in Rejiao village in southwestern Rutog from November 2005 through March 2006. PPR likely existed for several years without being recognized in Tibet because veterinarians, animal health workers, and livestock owners in the area are unfamiliar with its clinical and pathologic features. Also, this disease is frequently confused with other diseases that cause respiratory problems and death in small ruminants (The molecular epidemiologic techniques provided data suggesting cross-border transmission of PPRV infection into Tibet. PPR infection has been recognized in many Asian countries bordering southwestern China, including India (


Single cell suspensions of two allogeneic tumours (W-256 and Y-P388) injected intravenously produced macrocolonies in the lungs of rats. Colony forming efficiency  was low in 6-week or older rats but was markedly increased by 1000-1500 rad local thoracic irradiation (LTI) given 7-14 days before the tumour cell injection, or by antilymphocytic serum (ALS) but not by sublethal whole body irradiation (WBI). Similarly, LTI increased the incidence of pulmonary metastases produced by a solid tumour growing in the leg muscle. Stimulation of CFE by LTI was a strictly local phenomenon and not due to effects of irradiation on thymus, spleen or other tissues of the rat. LTI failed to increase CFE in immunized rats. It is concluded that (1) LTI stimulates clonogenic growth of tumour cells arrested in the lungs, by causing inflammatory reactions accompanied by regenerative cellular proliferation of lung tissue, which increases the “plating” efficiency of tumour cells, (2) the increase in CFE in lungs is not due to suppression of immunity to tumour growth by LTI.


The full spectrum technique in the fourier domain was studied forsimultaneous determination of Cu(II), Pb(II) and Cd(II) with 4-(2-pyridylazo) resorcinol (PAR). A program called SPGRFSQ. was designed to perform the calculations. Seven error functions were calculated for deducing the number of factors. Data reduction was performed using principal component analysis (PCA) algorithm. Experimental results showed the method to be successful even where there was severe overlap of spectra.


Knowledge and awareness about childhood autism is low among health workers and the general community in Nigeria and other Sub-Saharan African countries. Poor knowledge and awareness about childhood autism, especially among health workers can compromise early recognition and interventions which had been known to improve prognosis in children with autism. In formulating policy and designing interventions for these children, there is need to develop a reliable tool that can be used in assessing baseline knowledge about childhood autism among health workers and the impact that future continued education and awareness campaign may have on such baseline knowledge. Knowledge about childhood autism among health workers (KCAHW) questionnaire was designed for this purpose.The KCAHW questionnaire is a nineteen (19) item self-administered questionnaire that is divided into four domains. KCAHW questionnaires were distributed to fifty (50) psychiatric nurses involved in community mental health services in South-Eastern Nigeria to complete. After two weeks period, the KCAHW questionnaires were re-administered to the same fifty (50) psychiatric nurses to assess their knowledge about childhood autism and to assess the test-retest reliability and internal consistency of this questionnaire.KCAHW questionnaire showed good test-retest reliability when the mean domain and total scores at first and second time administration were compared. The four mean domain scores and the mean total scores at first and second time administration were significantly correlated. The questionnaire also had a good overall internal consistency when the mean scores of the four domains were correlated with mean total scores .The KCAHW questionnaire is a reliable tool for assessing knowledge of health workers about childhood autism. It would be a useful tool in improving early recognition of features of autism among affected children in Sub-Saharan African and other developing countries of the world where knowledge and awareness about childhood autism is low. Since Leo Kanner's paper titled, "Autistic disturbances of affective contact" about seven decades ago , awareneEarlier survey on knowledge about childhood autism among health care workers had been by Stone . The stuRhoades et al  exploredThe above surveys coming from the United States of America (USA), an environment where knowledge and research about childhood autism are more widely spread than what obtains in sub-Saharan Africa point to the need to assess knowledge and awareness in this environment and put in place programs to increase such knowledge and awareness with the ultimate aim of helping children with ASD.It had been presumed that in view of the rarity of presentation of childhood autism cases in pediatric health facilities in developing countries especially Sub-Saharan Africa, health care workers especially the nurses who are often in charge of various Primary Health Care (PHC) facilities in rural communities in Africa might not be well versed with the condition. Subsequently, early identification at the community level and intervention which have been shown to improve the prognosis of these cases ,21,22 maThis presumption was put into test in May 2007, when African Network for the Prevention and Protection Against Child Abuse and Neglect (ANPPCAN), Nigeria Chapter in collaboration with the World Bank carried out a survey to ascertain the level of knowledge and awareness of health care workers and the general public in Enugu State, South Eastern Nigeria on autism and implemented a project aimed at increasing the level of awareness on autism.The results of the survey showed that there is a very low level of awareness on autism among the general public and a low to average level of awareness among various categories of health care workers, with highest level of awareness noted among those health workers working in psychiatric facilities. Attempt was made then, to increase the level of awareness on autism through a workshop organized for the stakeholders and a radio phone – in program for the general public. However, one of the problems encountered during the survey was the unavailability of easily understandable and reliable questionnaire for measurement of the health care workers baseline knowledge of autism, aimed at early identification of cases in the community . The KCAIt had therefore been established that knowledge and awareness on autism and ASD is low in south-eastern Nigeria and there had been recent outcry in Nigeria to promote knowledge and awareness about childhood autism among the health care workers and the general community .In doing this, there is need for baseline assessment of present knowledge, facilities in place for intervention among other things. This would ensure proper planning. Promoting awareness at the community level should begin with various health workers who are involved in community health services. They can be trained to become trainers of other category of people in the community towards early recognition of children with childhood autism.For proper design of community campaign and continued education needed for these health workers, the need arises to design a reliable tool that can be used in measuring their baseline knowledge about childhood autism and the possible impact that future continued education and campaign may have on the knowledge of these health workers about childhood autism.Knowledge and awareness about childhood autism may vary from one geographical region to another. The same tool can be used to assess variation in knowledge of the disease condition over different geographical regions, given an index of geographical locations where immediate attention need to be focused as regards health workers and community education."Knowledge about Childhood Autism among Health Workers (KCAHW) Questionnaire" for use in various communities of developing countries where knowledge about childhood autism is limited among the health workers working at the community level.The above reasons inform the design of This article is aimed at describing the content of KCAHW questionnaire and reporting the test-retest reliability and internal consistency of the questionnaire.Location of the study was at Federal Neuro-Psychiatric Hospital, New Haven, Enugu (FNHE), Nigeria. FNHE is a specialist Psychiatric Hospital located in the South-Eastern region of Nigeria. The hospital is involved in community mental health service collaboration with various PHC facilities located in the rural communities of South-Eastern Nigeria. This community mental health care service in various PHC facilities in this region of the country is overseeing by the psychiatric nurses employed by the hospital.Ethical approval for this study was obtained from the ethical committee of FNHE, Nigeria.A total of fifty (50) psychiatric nurses employed by FNHE, Nigeria and who are involved in the community mental health services of the hospital participated in the study. There were 12 males and 38 females. The fifty nurses included in the study had had a minimum of five years of practice in general psychiatry nursing and possessed diploma qualification in general nursing and psychiatry nursing. It should be noted that only psychiatric nurses were used as the sample for this study because in the context of our environment in Nigeria and other sub-Saharan African countries, they are supposed to be the most likely health care workers who attend to cases such as autism and ASD and still supervise health care services at primary care level.In addition to the socio-demographic questionnaire, KCAHW questionnaire was distributed to consented fifty (50) psychiatric nurses to complete. After a period of two (2) weeks, the KCAHW questionnaire was re-distributed to the same fifty (50) psychiatric nurses to complete. The questionnaires were completed by the respondents and collected from them there and then at each point of administration to prevent them from consulting any study material or discussing with their colleagues before filling in their responses.This is a self administered questionnaire that contained a total of nineteen (19) – item questions. Each of the item questions has three (3) options to choose from with only one of these three options being correct. The correct option on each item question attracts a score of one (1), while the other two options that are incorrect attract a score of zero (0) each.The KCAHW questionnaire is further divided into four (4) domains:Contained eight (8) item questions that addressed the impairments in social interaction usually found in children with childhood autism. A maximum and minimum score of 8 and 0 respectively are possible in this domain.Contained only one (1) item question that addressed impairment in area of communication and language development, which is part of symptom presentation in children with childhood autism. A maximum and minimum score of 1 and 0 respectively are possible in this domain.Contained four (4) item questions that addressed area of obsession and compulsive pattern of behavior found in children with childhood autism, a pattern of behavior which had been described as restricted, repetitive and stereotyped. A maximum and minimum score of four (4) and zero (0) respectively are possible in this domain.Contained six (6) item questions that addressed information on what type of disorder is childhood autism, possible co-morbid conditions and onset of childhood autism in affected children. A maximum and minimum score of six (6) and zero (0) respectively are possible in this domain.Therefore, a maximum and minimum total score of nineteen (19) and zero (0) respectively are possible when the four domain scores are added together.The content of the questionnaire and the scoring system are shown in Appendices 1 and 2.The mean total score on KCAHW questionnaire among a particular sample population or community is a measure of level of knowledge about childhood autism among that particular sample population or community.The mean scores in each domain and the mean total scores generated when KCAHW questionnaires were administered the first and second time were compared with paired t-test statistical analysis to assess the test-retest reliability of the KCAHW questionnaire.Then, the mean inter-domain scores during the first and second time administration of the questionnaire were correlated with the mean total scores and cronbach's alpha  was calcA total of fifty (50) psychiatric nurses employed by FNHE, Nigeria and who are involved in the community mental health services of the hospital participated in the study. There were 12 males and 38 females with mean age of 32.6 years. The mean scores in the four domains of KCAHW questionnaire and mean total scores of 9.6 and 9.8 on first and second time administration of KCAHW questionnaire respectively which gave indices of level of knowledge on childhood autism in that particular sample population are shown in Table The paired t-test analysis comparing these mean scores and the 95% Confidence Interval (95% C.I) for the differences in the mean scores are shown in Table The correlation co-efficient for the four mean domain scores and mean total scores at first and second time administration of KCAHW questionnaire are shown in Table The internal consistency of KCAHW questionnaire was assessed by correlating different mean domain scores with the mean total scores at first and second time administration of the questionnaire and calculating the cronbach's alpha. The cronbach's alpha value of 0.97 obtained following the correlation is acceptable based on Nunnaly's recommendation .The ten by ten correlation matrix table of the four mean domain scores and mean total scores at first and second time administration of the KCAHW questionnaire is shown in Table The content of KCAHW questionnaire had been described. The test-retest reliability of the questionnaire is good based on the significant correlations in the mean domain and mean total scores at first and second time of administration. The KCAHW questionnaire also showed a good internal consistency as revealed in the cronbach's alpha value obtained for mean domain scores correlation with the mean total scores both at first and second time of administration.The imperative need to increase the level of knowledge and awareness among health workers and of the general community in Sub-Saharan African and other developing countries of the world about childhood autism necessitated the design of a reliable measuring tool that can be used to assess baseline knowledge of health workers on childhood autism and also subsequently the impact the future campaign and continuous education may have on the knowledge of these health workers about childhood autism, especially those group of health workers at primary health care facilities who are closer to the communities.KCAHW questionnaire could also be used to compare knowledge about childhood autism among different categories of health workers, if there is an indication for such. The variation in knowledge about childhood autism among health workers that could exist in various geographical regions of Sub-Saharan African and other developing countries where awareness about autism and ASD is low could also be assessed with KCAHW questionnaire.The limitation of administering the KCAHW questionnaire is that it could only be completed and collected immediately from the respondents upon administration. This is to prevent consultation of study materials or discussion with colleagues which can influence the respondents' response to the questions contained in the questionnaire. The KCAHW questionnaire would therefore serve the purpose only of a point time assessment of knowledge.KCAHW questionnaire is a reliable tool for assessing knowledge of health workers about childhood autism, aimed at early recognition of the condition in the primary care settings. It would be a valuable tool in assessing baseline knowledge of health workers about childhood autism in Sub-Saharan African communities and other developing countries where public awareness about childhood autism is low and policy and interventions are being planned for children affected by childhood autism. The level of knowledge of health workers about childhood autism, especially those working in the rural communities in this environment and other developing countries is an essential ingredient for early recognition and intervention which had been noted to improve prognosis in the affected children ,17,18. KPOE is affiliated with African Network for the Prevention and Protection against Child Abuse and Neglect (ANPPCAN), Nigeria Chapter and ANPPCAN, Nigeria Chapter received sponsorship from the World Bank in May 2007 for a survey on level of awareness of autism and implementation of program to raise awareness on autism in Enugu State, South Eastern Nigeria.All authors contributed to the conception of the study. MOB, POE and AOA are involved in writing and revision of the manuscript. All authors approved the final draft of the manuscript.Please do not consult formal text books to answer these questions.Thank you for your time.The following behaviors best describe a child with Childhood Autism:i. Marked impairment in use of multiple non-verbal behaviors such as eye to eye contact, facial expression, body postures and gestures during social interaction?(A) Don't Know, (B) Yes, (C) Noii. Failure to develop peer relationship appropriate for developmental age?(A) Don't Know, (B) Yes, (C) Noiii. Lack of spontaneous will to share enjoyment, interest or activities with other people? (A) Don't Know, (B) Yes, (C) Noiv. Lack of social or emotional reciprocity? (A) Don't Know, (B) Yes, (C) Nov. Staring into open space and not focusing on any thing specific?(A) Don't Know, (B) Yes, (C) Novi. The child can appear as if deaf or dumb? (A) Don't Know, (B) Yes, (C) Novii. Loss of interest in the environment and surroundings?(A) Don't Know, (B) Yes, (C) Noviii. Social smile is usually absent in a child with Autism?(A) Don't Know, (B) Yes (C) Noi. Delay or total lack of development of spoken language?(A) Don't Know (B) Yes (C) Noi. Stereotyped and repetitive movement (e.g. Hand or finger flapping or twisting)?(A) Don't Know (B) Yes, (C) Noii. May be associated with abnormal eating habit?(A) Don't Know, (B) Yes, (C) Noiii. Persistent preoccupation with parts of objects?(A) Don't Know, (B) Yes, (C) Noiv. Love for regimented routine activities? (A) Don't Know, (B) Yes, (C) Noi. Autism is Childhood Schizophrenia? (A) Don't Know, (B) Yes, (C) Noii. Autism is an auto-immune condition? (A) Don't Know, (B) Yes, (C) Noiii. Autism is a neuro-developmental disorder? (A) Don't Know, (B) Yes, (C) Noiv. Autism could be associated with Mental Retardation?(A) Don't Know, (B) Yes, (C) Nov. Autism could be associated with Epilepsy? (A) Don't Know, (B) Yes, (C) Novi. Onset of Autism is usually in, (A) Neonatal age, (B) Infancy, (C) Childhoodi Marked impairment in use of multiple non-verbal behaviors such as eye to eye contact, facial expression, body postures and gestures during social interaction?(A) 0 (B) 1 (C) 0ii Failure to develop peer relationship appropriate for developmental age?(A) 0 (B) 1 (C) 0iii. Lack of spontaneous will to share enjoyment, interest or activities with other people? (A) 0 (B) 1 (C) 0iv Lack of social or emotional reciprocity? (A) 0 (B) 1 (C) 0v Starring into open space and not focusing on any thing specific?(A) 0 (B) 1 (C) 0vi. The child can appear as if deaf or dumb? (A) 0 (B) 1 (C) 0vii. Loss of interest in the environment and surroundings?(A) 0 (B) 1 (C) 0viii. Social smile is usually absent in a child with Autism?(A) 0 (B) 1 (C) 0i. Delay or total lack of development of spoken language?(A) 0 (B) 1 (C) 0i. Stereotyped and repetitive movement (e.g. Hand or finger flapping or twisting)?(A) 0 (B) 1 (C) 0ii. May be associated with abnormal eating habit?(A) 0 (B) 1 (C) 0iii. Persistent preoccupation with parts of objects?(A) 0 (B) 1 (C) 0iv. Love for regimented routine activities? (A) 0 (B) 1 (C) 0i. Autism is Childhood Schizophrenia? (A) 0 (B) 0 (C) 1ii. Autism is an auto-immune condition? (A) 0 (B) 0 (C) 1iii Autism is a neuro-developmental disorder? (A) 0 (B) 1 (C) 0iv. Autism could be associated with Mental Retardation?(A) 0 (B) 1 (C) 0v. Autism could be associated with Epilepsy? (A) 0 (B) 1 (C) 0vi Onset of Autism is usually in (A) 0 (B) 0 (C) 1A total maximum score of 19 and a minimum score of 0 are possible. The average score on the KCAHW questionnaire among a particular sample population give an index level of knowledge about childhood autism in that particular population.


Elevated serum levels of the beta-subunit of human chorionic gonadotrophin (hCG) were measured in 50% of patients with locoregional recurrences or progressive vulvar carcinoma (n = 14). At diagnosis of vulvar cancer, however, the incidence of elevated serum levels was low (5%) in 104 patients. The rising serum levels during progression of disease indicate that the synthesis of the beta-subunit hCG can be increased in vulvar carcinoma.


In the northern hemisphere winter of 2003–04 antigenic variant strains (A/Fujian/411/02 –like) of influenza A H3N2 emerged. Circulation of these strains in the UK was accompanied by an unusually high number of laboratory confirmed influenza associated fatalities in children. This study was carried out to better understand risk factors associated with fatal cases of influenza in children.Case histories, autopsy reports and death registration certificates for seventeen fatal cases of laboratory confirmed influenza in children were analyzed. None had a recognized pre-existing risk factor for severe influenza and none had been vaccinated. Three cases had evidence of significant bacterial co-infection. Influenza strains recovered from fatal cases were antigenically similar to those circulating in the community. A comparison of protective antibody titres in age stratified cohort sera taken before and after winter 2003–04 showed that young children had the highest attack rate during this season . Clinical incidences of influenza-like illness (ILI) in young age groups were shown to be highest only in the years when novel antigenic drift variants emerged.This work presents a rare insight into fatal influenza H3N2 in healthy children. It confirms that circulating seasonal influenza A H3N2 strains can cause severe disease and death in children in the apparent absence of associated bacterial infection or predisposing risk factors. This adds to the body of evidence demonstrating the burden of severe illness due to seasonal influenza A in childhood. Influenza A virus is a cause of severe morbidity and mortality worldwide Human seasonal influenza A viruses evolve rapidly through genetic mutation, allowing them to escape previously acquired immunity This study investigated fatal cases of influenza H3N2 in children during the emergence of A/Fujian/411/02 in the UK in winter 2003–04. The goals of the study were: to investigate whether any of the case fatalities had underlying disease or risk factors; to evaluate how laboratory confirmed infections were recorded in death certification; to describe the clinical and pathological findings in relation to circulating virus strains; and to investigate whether the high number of laboratory confirmed fatalities was reflected in measures of morbidity in primary care, age specific attack rates and age specific all-cause mortality. We reviewed the antecedent illness history, underlying risk factors, clinical findings and observed pathology at autopsy. Age-stratified serological data were combined with community based clinical morbidity data to provide an assessment of the overall population immune susceptibility, attack rates and impact of A/Fujian/411/02-like influenza in comparison with other winter seasons in which new H3N2 antigenic variants emerged.This study was conducted as part of a public health inquiry by the Health Protection Agency and Health Protection Scotland into paediatric deaths, where it was agreed that ethical approval for linking personal clinical outcome information to virus strains was not required. Nationally representative, anonymised, age stratified residual sera were obtained from the HPA serum bank held in Preston NHS hospital. Ethical approval was given by the HPA for the collection of these samples.st September 2003 to 30th May 2004) with laboratory confirmation of influenza virus infection reported to HPA. Most specimens were initially tested at referring hospitals, and all were subsequently tested at the national influenza laboratory, HPA Centre for Infection, Colindale (HPA CfI). Microbiological findings deemed significant are included.A case was defined as a death in child less than eighteen years of age during the 2003–04 influenza season . The activity took place within the context of a national public health enquiry into influenza associated childhood deaths in winter 2003. Children without risk factors recognized by UK Joint Committee on Vaccination and Immunisation to increase the risk of influenza related complications were classified as “previously healthy”. U.K. risk factors for severe influenza from 2003–04 are given in reference 12. Verification of the certified cause of death in each fatal case was performed by matching cases to Office of National Statistics (ONS) death registration records held at HPA.http://www.hpa.nhs.uk/web/HPAweb&HPAwebStandard/HPAweb_C/1195733836222.The Weekly Returns Service (WRS) of the Royal College of General Practitioners is a clinical information system based upon a network of sentinel general practices located throughout England and Wales Viruses were grown in either MDCK or MDCK-SIAT1 cells and subtyping was performed by RT-PCR, if no virus isolate was available Nationally representative, anonymised, age stratified residual sera were obtained with ethical approval from the HPA serum bank held in Preston NHS hospital. The number of sera tested from 2003 was 88 (ages 0–4), 196 (ages 5–17), 293 (ages 18–44), 185 (ages 45–64) and 101 (ages 65+). The number of sera from 2004 for each age group was 70 (ages 0–4), 160 (ages 5–17), 273 (ages 18–44), 199 (ages 45–64) and 98 (ages 65+).Haemagglutination inhibition (HI) assays to assess protective antibody titres in human sera were performed using standard protocols and antigenically representative viruses. A titre of >1/40 was taken as protective.Seventeen fatal cases of laboratory-confirmed influenza A in children under eighteen years were reported to HPA between September and December 2003. This period corresponded with peak circulation of influenza during the winter 2003–04. The median age of fatal cases was two years, with a range from four months to seventeen years; the majority (64%) were under five years. Ten (58%) deaths were in females. The duration of illness prior to death varied, with a median duration of three days (range 0–14 days). Four (24%) deaths were sudden with either no recognized antecedent illness or illness of less than 6 hours duration. A further four (24%) occurred within forty-eight hours of onset of symptoms, five (29%) were ill for three to four days prior to death, three (18%) were ill for a week prior to death, and one individual was unwell for over ten days. Eight (47%) children died following hospitalization and nine (53%) died in the community. In the cases with recognized antecedent illness, eight (57%) had fever, eleven (79%) had respiratory symptoms (cough/chest infection/croup), three (21%) had vomiting and/or diarrhea, two (14%) had clinically recognized sepsis. Two (14%) children were markedly confused prior to collapse .Among the sixteen cases for whom underlying health status information was available none was recorded as having a known risk factor for severe influenza, although two cases (both over two years) had a history of febrile convulsions In nine cases (53%) influenza A detection occurred by more than one method , either in respiratory clinical samples or from post-mortem tissues. In eleven (64%) cases influenza A virus isolates were recovered. The cultured viruses were analyzed for their antigenic characteristics; all isolates were found to be antigenically similar to each other and to virus isolates recovered from non-fatal cases matched for age, week of infection and geographical location (controls) as well as to other circulating strains from the same locality (data not shown). Influenza AH3N2 was detected by RT-PCR only in five (29%) cases.Staphylococcus aureus and Streptococcus pneumoniae in blood were identified in the two cases of clinical sepsis and one case had Streptococcus pneumonia bacterial pneumonia. No other significant bacteriological findings were described. Enterovirus and norovirus were also detected by RT-PCR in two cases.Staphylococcus aureus or Streptococcus pneumoniae and myocarditis in two cases (12%). Eight cases (47%) showed abnormalities of the lymphoid system, including seven (41%) with enlargement of the lymph nodes (LN), thymus or spleen, six (35%) with hemorrhagic LN, thymus or spleen and two (12%) with “starry sky” LN indicating an infiltration of macrophages in the lymphoid tissue. No metabolic disorders were recognized ante or post mortem, although one case showed unusual large fiber laryngeal musculature of uncertain significance.Autopsy reports were available for all cases . All casDeath registration information was not available for one case. By routine death registration only nine of sixteen fatal cases (56%) were coded with influenza (ICD10 J10–J11). Seven of these specifically mentioned influenza A on the death certificate and a further certificate mentioned “influenza” when text was searched manually. Two (13%) of the deaths were registered as pneumonia (J18); two (13%) as other respiratory disorders (J98); two (13%) were registered as sudden infant death (SIDS) and one as acute myocarditis, without mention of influenza (I40). Septicemia (A41) and myocarditis  were identified as contributing causes to death on three certificates. No death certificate indicated any chronic medical conditions known to increase the risk of severe disease from influenza infection.We sought to establish whether young children showed particular susceptibility to A/Fujian/411/02-like virus infection during this winter season by analyzing sera from summers 2003 and 2004, prior and subsequent to the emergence of this antigenic drift variant. Serum samples in a nationally representative age-stratified serum cohort were analyzed for the presence of protective antibodies . The numTo assess the morbidity of influenza in different age groups we analysed observational clinical data from primary care recorded by GPs of the Weekly Returns Service (WRS). This is a national sentinel network which has been recording influenza-like illness (ILI) for over 40 years, and for which linked clinical-virological surveillance has been operational since 1994. The all-age incidences of ILI in England and Wales over a ten year period from 1994–2004, together with incidence rates in children ages 0–4 and 5–14 years is shown in Winter 2003–04 was marked by an unusually high number of severe and fatal cases of influenza in the young, following the emergence of the influenza A H3N2 antigenic drift variant A/Fujian/411/02 The range of clinical complications in UK fatal cases can be grouped into categories including (1) fulminant progression to death after an initially mild illness, (2) invasive bacterial infection, (3) respiratory tract complications and (4) non respiratory complications including myocarditis and encephalopathy. Extrapulmonary complications of myocarditis and encephalopathy were not accompanied by the detection of viral RNA from brain or heart tissue by RT-PCR, although virus was recovered from the nasopharynx. The pathological conclusions rest on histology findings, which suggests that tissue damage may occur through mechanisms other than direct viral replication, as has been previously suggested Staphylococcus aureusThe clinical microbiological findings add to the debate about the importance of bacterial co-pathogens in influenza associated fatality. Only three (18%) of the fatal cases showed evidence of significant bacterial co-infection. This is similar to findings from fatal cases in the US 2003–04 and also to preliminary information from the current outbreak of swine-origin influenza A H1N1, where a minority of paediatric hospital admissions had possible or probable bacterial infection In this case series over 40% of death certificates had no mention of influenza as a direct or indirect cause of death, and in over 70% of cases the diagnosis of influenza was not made until post mortem tissue was examined. The burden of influenza in young children is therefore under recognized, precisely because few influenza infections are recognized clinically A risk-factor based influenza vaccination program for children would not prevent these fatal cases as the reasons underlying susceptibility to severe disease remain cryptic. Further studies on the outcome of seasonal influenza in children will help us to predict the impact of future epidemics and will assist understanding of the outcome of infections in the immune naïve host during influenza pandemics.


The differential p53 overexpression in different subtypes of intestinal metaplasia and gastric cancer suggests that type III intestinal metaplasia is the commonest lesion in dysplasia-carcinoma transition, particularly in the intestinal type of gastric cancer.


Higher concentrations of PSA and hK2 were found in tumours expressing steroid hormone receptors , and both PSA  and hK2  correlated directly with PR levels. A negative correlation between patient age and PSA  was also found. Both proteins were present in nipple aspirate fluid at relatively high concentrations which were positively correlated . The molecular weights of the immunoreactive species quantified by the hK2 and PSA assays were established by high-performance liquid chromatography (HPLC) and were consistent with the known molecular weights of hK2 and PSA. Together these data provide the first evidence, to our knowledge, that both malignant breast tissue and normal breast secretion contain measurable quantities of hK2, and that the degree of hK2 expression or secretion is directly proportional to the expression of PSA and steroid hormone receptors. hK2 expression may therefore be a marker of steroid hormone action in breast tissue. © 2000 Cancer Research CampaignThe recent demonstration of human glandular kallikrein (hK2) expression in a breast carcinoma cell line has suggested that this putatively prostate-restricted, steroid hormone-regulated protease may also be expressed in breast epithelium in vivo and secreted into the mammary duct system. Given that the only substrate yet identified for hK2 activity is the precursor of prostate-specific antigen (PSA), the expression of which in breast carcinomas may be associated with favourable prognosis, our purpose was to examine the expression pattern of both hK2 and PSA in breast tumour tissues. Cytosolic extracts of 336 primary breast carcinomas prepared for routine oestrogen receptor (ER) and progesterone receptor (PR) analysis, as well as 31 nipple aspirates from six women with non-diseased mammary glands, were assayed for hK2 and PSA using immunofluorometric assays developed by the authors. In the tumour extracts, measurable hK2 and PSA concentrations were detected in 53% and 73% of cases respectively, and were positively correlated to each other (


DCC is the most likely target of LOH because loss or reduction of DCC expression has been found in many types of cancers. However, few reports have focused on sequence mutations of this gene. We investigated sequence mutations and expression of DCC in primary gastric cancers. We studied mutations in 25 of the 29 DCC exons by PCR-SSCP in 17 primary gastric cancers exhibiting LOH on 18q21. No mutations of DCC were found in any of the tumours, although 78% (47/60) of the primary tumours showed apparent loss or reduction of DCC expression by immunohistochemistry. Analysis of methylation status of DCC revealed that methylation frequently occurred in both primary tumours  and corresponding non-cancerous gastric mucosae . Methylated status of DCC was significantly correlated with the loss of DCC expression in primary tumours (P< 0.01). These results indicate that DCC is frequently silenced, probably by epigenetic mechanisms instead of sequence mutations in gastric cancer. © 2001 Cancer Research Campaign http://www.bjcancer.comLoss of heterozygosity (LOH) on chromosome 18q21 is frequently found in various human cancers, suggesting the presence of tumour suppressor gene(s) in this chromosomal region.


To identify and implement strategies that help meet safety monitoring requirements in the context of an observational study for artemether-lumefantrine (AL) administered as first-line treatment for uncomplicated malaria in rural Tanzania.Pharmacovigilance procedures were developed through collaboration between the investigating bodies , the relevant regulatory authority (Tanzania Food and Drugs Authority) and the manufacturer of AL (Novartis Pharma AG). Training and refresher sessions on the pharmacovigilance system were provided for healthcare workers from local health facilities and field recorders of the Ifakara Health Demographic Surveillance System (IHDSS). Three distinct channels for identification of adverse events (AEs) and serious adverse events (SAEs) were identified and implemented.st September 2007 to 31st March 2010, 57 AEs were reported . The remaining seven were due to anti-pyretic or anti-parasite medications. Twenty patients experienced SAEs; in 16 cases, a relation to AL was suspected. Six of the 20 cases were reported within 24 hours of occurrence. In all cases reported after AL, 5 cases (vomiting), 16 cases (itching and/or rash) and others were difficult breathing, convulsion and headache (10 cases). The four AEs that occurred after SP were mild erythematic skin lesions that did not progress to Stevens-Johnson syndrome. The AEs seen following treatment with penicillin and amoxicillin were both rashes.Training was provided for 40 healthcare providers (with refresher training 18 months later) and for six field recorders. During the period 1Safety monitoring and reporting is possible even in settings with weak health infrastructure. Reporting can be enhanced by regular and appropriate training of healthcare providers. SMS text alerts provide a practical solution to communication challenges.Experience gained in this setting could help to improve spontaneous reporting of AEs and SAEs to health authorities or marketing authorization holders.


Fibreoptic bronchoscopy in previously untreated patients with malignant lymphoma provided diagnostic information in 8 of 25 cases with radiological evidence of intrathoracic involvement. There was a marked difference in the pattern of endobronchial involvement between Hodgkin's Disease (HD) and Non Hodgkin's Lymphoma (NHL) which specifically infiltrated the bronchus-associated lymphoid tissue. Bronchoalveolar lavage was abnormal in only one patient with Hodgkin's Disease in whom the presence of many Sternberg-Reed cells suggested occult dissemination of otherwise localised disease.


One hundred and seven patients with biliary pancreatitis undergoing operation from 1976 to 1989 werereviewed. To clarify the reason for failure to respond to conventional supportive therapy, 73 patients (68%) who underwent emergency surgery were retrospectively divided into two groups according to the severity of the pancreatitis evaluated at laparotomy and compared. Sixty-two had minimal or mild pancreatitis (Group I), among whom 44 (71%) had life-threatening acute biliary tract disease. All underwent biliary surgery and 4 (6%) subsequently died, 2 due to acute obstructive suppurative cholangitis. Eleven had hemorrhagic necrotizing pancreatitis (Group II), among whom 7 had complications of acute pancreatitis such as pancreatic ascites or abscess. These underwent pancreatic and/orbiliary surgery and 3 (27%) died of multi-organ failure.		There appears to be two types of biliary pancreatitis refractory to conventional supportive therapy, which differ in the extent of surgery required and in mortality: (1) minimal or mild pancreatitis with persistent life-threatening acute biliary tract disease (biliary type), and (2) more severe pancreatitis (pancreas type) early in the course of the disease.


The analyses were carried out with the aid of ion-exchange chromatography. The molar proportions of 5 bases detected in acid hydrolysates of tumour tRNA (N


The measurement of carcinoembryonic antigen (CEA) in serum and endoscopic brush specimens was evaluated for the differential diagnosis of malignant and nonmalignant gastric disease. Brush specimens were studied from 33 patients with gastric cancer and 36 patients with benign gastric lesions or apparently normal gastric mucosa. Demonstrable CEA immunoreactivity was found by radioimmunoassay in brush specimens from 24/33 cancer patients (73%) and from 23/36 patients with benign lesions (64%). Patients with CEA+ tissue in the immunoperoxidase test had somewhat higher CEA concentrations in the brush specimens than cases with CEA- biopsy tissue, although overlap was considerable. Thirty-five per cent of cancer patients had both a positive tissue CEA reaction and a CEA/DNA ratio greater than 10 ng/micrograms, whilst patients with benign lesions had only 15% of positives by these criteria (0.01 greater than P greater than 0.001). The serum CEA concentration was above the upper normal level of 5 ng/ml in 2/39 patients, both of whom had gastric cancer. The CEA immunoreactive material from benign and malignant lesions eluted in gel filtration on Sephadex G-200 in the same volume as CEA purified from liver metastases of cancer of the colon, showing that a glycoprotein sharing immunological and physicochemical properties with CEA is present both in malignant and nonmalignant lesions of the gastric mucosa, and that there is considerable overlapping in the amount of CEA. The estimation of CEA in gastric-brush specimens is therefore of limited value in the differential diagnosis of benign and malignant gastric lesions.


Peripheral blood leucocytes from chronic myeloid leukaemia patients in remission were tested for inhibition of migration in presence of solubilized membrane antigens from leukaemic cells in 15 cases. Eight out of 9 autochthonous combinations (88-8%) and 35/49 allogenic combinations (71-4%) showed inhibition of migration. Antigens prepared from relapse leukaemic cell samples in 4 cases showed inhibition of migration of autochthonous as well as allogeneic remission leucocytes. The same batch of CML antigens inhibited migration of normal leucocytes at the level of 22-2%. The difference between inhibition of migration shown by remission leucocytes and normal leucocytes in presence of CML antigens was statistically significant. Solubilized antigens, similarly prepared from normal leucocytes, showed inhibition of migration of remission leucocytes to the extent of 15% only. The difference between the reactivity of CML remission leucocytes to normal and CML antigens was also statistically significant. No enhancement of migration of remission leucocytes was seen with CML antigens.


The macrophage electrophoretic mobility test described by Caspary and Field (1971) and modified by Pritchard et al. (1973) was investigated in various models of cell-mediated immune conditions in the guinea-pig and in cancer in man. No positive results were obtained in 92 guinea-pig experiments. Only 17 of 154 experiments on 74 patients gave definite positives in experiments with human cancer and a few positive results were obtained with normal healthy subjects.


Forty-four (44) patients with Burkitt's lymphoma received identical combination chemotherapy on the basis of body surface area. Patients with renal dysfunction, more common in those with abdominal tumours, were at significantly greater risk of developing severe leucopenia (less than 1000 cells/dl) than those with normal renal function (P less than 0.0001). Similar results were seen in a series of 8 patients with normal marrows treated with only i.v. cyclophosphamide and intrathecal methotrexate. Giving a lower initial dose of cyclophosphamide seemed to reduce the risk of severe leucopenia in 5 additional patients with evidence of renal dysfunction. The mechanism is postulated as delayed excretion of the active metabolites of cyclophosphamide. Adjustment of the chemotherapeutic dose should be considered when treating patients with renal dysfunction.


We compared antimicrobial resistance patterns of Salmonella enterica serovar Typhimurium (ST) of isolates from humans (n = 715) and cattle (n = 378) in the Pacific Northwest from 1982 through 1997. The major changes in antimicrobial resistance can be attributed to the widespread clonal dissemination of multidrug-resistant definitive phage type 104 ST.


Twenty-seven patients with malignant neoplasia were injected intradermally with the methanol extraction residue (MER) fraction of tubercle bacilli. Two schedules of treatment were used: every other week and once a month; 1-10 courses of MER were administered to the patients. The skin reactivity to 3 recall antigens, as well as to the injected MER itself, was used to monitor the immune response. Improvement of skin reactivity occurred in 9 of 18 patients tested with recall antigens. Five of 6 patients treated every other week improved in their immune capacity whereas only 4 of 12 patients improved on the monthly schedule. Thus, repeated injections given every other week were more effective in increasing the cutaneous reactivity than monthly injections of MER. The side-effects of MER treatment were tolerable.


High levels of loss of distal markers on 17p13.3 in breast cancer suggested the presence within the region of at least one tumour-suppressor gene. Here we describe the derivation of two biallelic polymorphisms from the 17p telomeric yeast artificial chromosome (YAC) TYAC98. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex PCR analysis demonstrated that the high level of allelic imbalance observed in breast tumours represented loss of constitutional heterozygosity (LOH) and that this LOH extended to the telomere. Lung carcinoma (but not Wilms' tumour)-derived DNA again revealed a high level of loss of subtelomeric 17p sequences. Telomeric microsatellite polymorphisms from other chromosome arms did not show such elevated loss in either tumour type. This suggested that the 17p loss observed did not reflect a general telomeric instability and provided further evidence for the presence of a breast cancer tumour-suppressor gene in the distal region of 17p13.3.


They co-evolved into an intricate splicing code together with additional functional constraints, such as tissue-specific and alternative splicing patterns. We used orthologous exons extracted from the University of California Santa Cruz multiple genome alignments of human and 22 Tetrapoda organisms to predict candidate enhancers and silencers that have reproducible and statistically significant bias towards annotated exonic boundaries.Auxiliary splicing sequences play an important role in ensuring accurate and efficient splicing by promoting or repressing recognition of authentic splice sites. These Tetrapoda enhancers and silencers were clustered into 15 putative core motifs based on their Markov properties. Most of these elements have been identified previously, but 118 putative silencers and 260 enhancers (~15%) were novel. Examination of previously published experimental data for the presence of predicted elements showed that their mutations in 21/23 (91.3%) cases altered the splicing pattern as expected. Predicted intronic motifs flanking 3' and 5' splice sites had higher evolutionary conservation than other sequences within intronic flanks and the intronic enhancers were markedly differed between 3' and 5' intronic flanks.A total of 2,546 Difference in intronic enhancers supporting 5' and 3' splice sites suggests an independent splicing commitment for neighboring exons. Increased evolutionary conservation for ISEs/ISSs within intronic flanks and effect of modulation of predicted elements on splicing suggest functional significance of found elements in splicing regulation. Most of the elements identified were shown to have direct implications in human splicing and therefore could be useful for building computational splicing models in biomedical research. Eukaryotic genes contain intervening sequences or introns that need to be removed from precursor messenger RNA (pre-mRNA) in a complex process termed splicing. During pre-mRNA splicing, relatively short exonic sequences are recognized by spliceosome, a large RNA-protein complex. During splicing, introns are removed and exons are joined together to form mature RNA. In addition to splice site (SS) signals at the exonic 5' and 3' ends, accurate discrimination of exons and introns requires additional auxiliary elements -3. Thesein vivo or in vitro selection methods have been employed to identify these motifs in the genomic sequences. For example, the RESCUE-ESE (Relative Enhancer and Silencer Classification by Unanimous Enrichment), a computational approach used in conjunction with experimental validation, predicted specific hexanucleotide sequences as candidate ESEs based on significantly higher frequency of occurrence in exons than in introns and also significantly higher frequency in exons with weak SSs than in exons with strong SSs  we have found 88 5'SS-related and 330 3'SS-related ISEs/ISSs [see Additional File Tetrapoda superclass  3'SS and 177 (6.95%) 5'SS supporting motifs are novel and have not been previously reported in systematic screens detecting such elements. Among our predicted elements, 51.81% were octamers and 41.08% of sequences were heptamers as compared to only 6.76% hexamers and 0.35% pentamers, suggesting that motifs of larger size play important role in splicing regulation. We detected intersections with some of the cis-acting elements reported in the previous studies, but not nearly as dramatic as we saw between the intronic elements predicted for primates and Tetrapoda non-eutherian (an outgroup) clades. It demonstrates high reproducibility of our results obtained for various vertebrate lineages and supports the existence of highly conserved splicing regulatory code across vertebrates. This result also suggests the implications of elements found in regulating human splicing and may help explaining human hereditary disorders caused by mutations modulating such elements. We have established the higher evolutionary conservation for the predicted intronic cis-acting elements within mammalian intronic flanks which indicates their functional significance in exon definition. The elements found contain many of the known cis-acting factor binding sites with functionality supported by experiments with splicing reporter constructs. All these lines of evidence suggest active involvements of the predicted elements in control gradient directing spliceosome to the proper exons in the process of pre-mRNA splicing [Using the orthologous exons currently available for 23 cis-acting elements with the secondary pre-mRNA local structure in the vicinity of the SSs, except for slightly increased single strandedness detected for 5' and 3' ISSs deep inside introns. This observation is in contrast to the earlier reported [We did not observe statistically significant association for the predicted groups of reported , where kA remarkable intersection between the 5'SS ISSs and the 5'SS ESSs [see Additional File [Homo sapiens), Chimpanzee (Pan troglodytes), Rhesus (Macaca mulatta), Mouse (Mus musculus), Rat (Rattus norvegicus), Rabbit (Oryctolagus cuniculus), Dog (Canis familiaris), Cow (Bos taurus), Armadillo (Dasypus novemcinctus), Elephant (Loxodonta africana), Tenrec (Echinops telfairi), Opossum (Monodelphis domestica), Chicken , Frog . The "threaded blockset alignments" [ against reference human chromosomal assemblies  using the BLAT program [We parsed and extended blocks of orthologous with human reference exons from multiple sequence alignment of 17 vertebrate genomes obtained from UCSC genome browser . The folgnments" , built u program . Having  program ,35 and d program  to make Otolemur garnetti), Tree Shrew (Tupaia belangeri), Guinea Pig (Cavia porcellus), Shrew (Sorex araneus), Hedgehog (Erinaceus europaeus), Cat (Felis catus), Horse , Platypus (Ornithorhynchus anatinus), Lizard (Anolis carolinensis). The blocks from a total of 28 vertebrates multiple genome alignments are normally shorter than blocks from 17 vertebrates multiple genome alignments, therefore chances are higher that the block extension may not produce the correct exonic boundaries. Only the exons associated with the species not obtained through the first round should be processed in the second round.This exon set was extended with exons derived from processing of 28 vertebrates multiple genome alignments obtained from UCSC genome browser  from theTetrapoda organisms. For these purposes we have examined 489,668 extended exons in primates clade (Human (Homo sapiens), Chimpanzee (Pan troglodytes), Rhesus (Macaca mulatta), Bush Baby (Otolemur garnetti)) and 476,218 extended exons from non-eutherian Tetrapoda organisms (Opossum (Monodelphis domestica), Chicken , Frog , Platypus (Ornithorhynchus anatinus), Lizard (Anolis carolinensis)) taken as the most distant outgroup  among Tetrapoda organisms relative to primates.To establish the firm ground for using sequences from distantly related organisms in predicting common SS proximal elements and to estimate implication of elements found in modeling human splicing we conducted independent search for the elements in two distantly related clades of primates and non-eutherian Homo sapiens), Chimpanzee (Pan troglodytes), Rhesus (Macaca mulatta), Bush Baby (Otolemur garnetti)) and rodents (Mouse (Mus musculus), Rat (Rattus norvegicus), Guinea Pig (Cavia porcellus)) clades.To estimate the increased conservation of the intronic elements found within the intronic flanks we have used the Prank  tool to Through the literature search we collected the test set of 185 human genes previously linked to autism spectrum disorder and genes implied in environmental response . A set odepth correspond to the density of an oligonucleotide of size depth present at a certain positions within genome. A significant deviation of an oligo density in the proximity of SSs as compare to background locations is strongly indicative of important functionality of elements related to splicing [We measured a statistically significant bias for all the possible oligonucleotides of size equal or less than 8 bp in the vicinity of true 5' and 3' SSs compared to distant "background" locations as shown in Figures splicing ,15,33.α = 0.011 using χ2 test involved 87,380 statistical hypotheses testing for all possible oligos of size less or equal to 8 bp located at certain position relative to a SS. Following the Bonferroni correction for multiple hypothesis testing we reduced the significance level to χ2 tests for the SS vicinity counts ≥ 63 or deep intronic/exonic counts ≥ 63 are statistically significantComparing oligo counts at the significance level of under conditionsorSkip value 29 nt, as shown in Figure Skip = 0 nt in Figure Skip = 29 nt in Figure Comparative measurements between the regions shown in Figure Extended orthologous exons associated with the same multiple genome alignment block frequently contain identical conserved oligonucleotides at certain positions relative to SSs, especially within exons. These conserved elements violate our assumption of independence of the sequences used for analysis of statistical significance; therefore within a block we counted unique element at certain position relative to SSs only once, disregarding all other identical motifs conserved since speciation from a common ancestor. To prevent substantial element underscoring in a counting round, since many of the elements at a certain position relative to a SS were sorted out due to evolutionary conservation, we shifted an element position by one within a counting region for each consecutive sequence as shown in Figure where mod is a modulo operation, counting round could be 0,1 or 2 and the sequence index goes from 1 to 2,333,379. Elements shifted by one position within the region are normally different and therefore not sorted out for being similar, which allows combining more elements in the region associated with a block under the same evolutionary pressure.These predicted groups of elements were clustered with the Mixture of Hidden Markov Models (MHMM), an unsupervised clustering method capable of modeling dependencies between neighboring positions in active motif cores [see Additional File . To accelerate finding average PU value for an element we calculated them only for the contexts of 11, 15, 20, 25 and 30 nt according to [The PU values for the predicted octamers surrounded by ± 30 nt context were calculated as described in  using RNrding to  were makAC designed and implemented the framework to search for splicing regulatory elements and tested the application. CH initiated the study, provided many valuable suggestions steering the project and was the head of the lab where the work was done. IV provided necessary expertise to formulate the study objectives, obtained experimental data to support newly found elements and extensively edited the manuscript. All authors read and approved the final manuscript.Report and analysis of splicing regulatory elements found. Contains list of all the elements found along with their statistical significance and overlap between their groups. Functional analysis for these elements in context of alternative splicing constructs is reported. We provided brief description for an unsupervised clustering method used to group the elements found and grouped the elements within possible functional motifs. We also report elements found for primates and outgroup. Subset of novel elements is reported.Click here for file


Paraffin wax embedded formalin-fixed benign breast disease tissue taken from 17 patients (15 with microcystic disease and 2 with fibroadenoma) was studied for the presence of tissue bound prolactin using a rabbit antiserum against human prolactin applied in conjunction with a highly sensitive modified version of the dinitrophenyl (DNP)-hapten sandwich staining (DHSS) procedure. Sections taken from 14 of the 15 cases showing apocrine cystic changes exhibited strong prolactin staining restricted to the cytoplasm of metaplastic apocrine cells lining the cysts. Normal lobules and ducts and blunt duct proliferations were all negative, as were also the two cases of fibroadenoma. In contrast 6 out of 8 cases of breast cancer examined showed heterogenously distributed cytoplasmic staining in the cancer cells. Maximal prolactin staining in the apocrine cells was observed at antiserum dilutions as high as 1:60,000. This compared favourably with a 1:120,000 dilution that gave maximal levels of staining in the prolactotrophs present in serial sections taken from formalin fixed paraffin wax embedded post mortem human anterior pituitaries. In both types of tissues the specific staining was abolished by pre-absorption of the antiserum with human prolactin (10 micrograms ml-1). No staining was observed when the anti-prolactin serum was either omitted or substituted with DNP-labelled normal rabbit serum. Apocrine metaplasia in cystic disease of the breast has recently been found to be associated with an increased breast cancer risk. The strong and selective presence of immunohistochemically demonstrable prolactin in the metaplastic cells may be of significance in view of the hormone's known growth stimulating effect on the breast epithelium.


Environ Health Perspect 117:A150–A156 (2009)], a statement reads, “The primary global impact of oil sands comes through the release of greenhouse gases created when about 800 million cubic feet of natural gas  is burned daily to create heat for extraction and upgrading, says Stringham.” To clarify: This statement refers to the global impact of oil sands production. As noted in the preceding paragraph of the article, most of the greenhouse gas emissions related to oil sands overall come from burning the fuel derived from these reserves.On p. A155 of the April Focus article [