Identification and initial characterization of matrix metalloproteinases in the yellow fever mosquito, Aedes aegypti.

Aedes aegypti is a major vector for arboviruses such as dengue, chikungunya and Zika viruses. During acquisition of a viremic bloodmeal, an arbovirus infects mosquito midgut cells before disseminating to secondary tissues, including the salivary glands. Once virus is released into the salivary ducts it can be transmitted to another vertebrate host. The midgut is surrounded by a basal lamina (BL) in the extracellular matrix, consisting of a proteinaceous mesh composed of collagen IV and laminin. BL pore size exclusion limit prevents virions from passing through. Thus, the BL probably requires remodelling via enzymatic activity to enable efficient virus dissemination. Matrix metalloproteinases (MMPs) are extracellular endopeptidases that are involved in remodelling of the extracellular matrix. Here, we describe and characterize the nine Ae. aegypti encoded MMPs, AeMMPs 1-9, which share common features with other invertebrate and vertebrate MMPs. Expression profiling in Ae. aegypti revealed that Aemmp4 and Aemmp6 were upregulated during metamorphosis, whereas expression of Aemmp1 and Aemmp2 increased during bloodmeal digestion. Aemmp1 expression was also upregulated in the presence of a bloodmeal containing chikungunya virus. Using polyclonal antibodies, AeMMP1 and AeMMP2 were specifically detected in tissues associated with the mosquito midgut.

Fibrin-Targeting Immunotherapy for Dementia.

Blood-brain barrier (BBB) disruption is an early event in the development of Alzheimer's disease. It precedes extracellular deposition of amyloid-ß in senile plaques and blood vessel walls, the intracellular accumulation of neurofibrillary tangles containing phosphorylated tau protein, microglial activation, and neuronal cell death. BBB disruption allows the coagulation protein fibrinogen to leak from the blood into the brain, where it is converted by thrombin cleavage into fibrin and deposits in the parenchyma and CNS vessels. Fibrinogen cleavage by thrombin exposes a cryptic epitope termed P2 which can bind CD11b and CD11c on microglia, macrophages and dendritic cells and trigger an inflammatory response toxic to neurons. Indeed, genetic and pharmacological evidence demonstrates a causal role for fibrin in innate immune cell activation and the development of neurodegenerative diseases. The P2 inflammatory epitope is spatially and compositionally distinct from the coagulation epitope on fibrin. Mouse monoclonal antibody 5B8, which targets the P2 epitope without interfering with the clotting process, has been shown to reduce neurodegeneration and neuroinflammation in animal models of Alzheimer's disease and multiple sclerosis. The selectivity and efficacy of this anti-human fibrin-P2 antibody in animal models supports the development of a monoclonal antibody drug targeting fibrin P2 for the treatment of neurodegenerative diseases. THN391 is a humanized, affinity-matured antibody which has a 100-fold greater affinity for fibrin P2 and improved development properties compared to the parental 5B8 antibody. It is currently in a Phase 1 clinical trial.

Epidemiology of renal cell carcinoma in Connecticut, 1935-1973.

Data on over 3,700 patients with renal cell carcinoma, reported to the Connecticut Tumor Registry from 1935 through 1973, were used to assess incidence, survival, and associations of risk with demographic characteristics. Incidence increased over time among men, but not among women; a birth cohort effect suggesting increasing incidence rate over time was demonstrated for men. A comparison of male and female age-specific incidence rates indicated that, in the 15- to 39-year-old age group, men were three times more likely than women to develop the disease; after age 40, renal cell carcinoma was diagnosed in men twice as often as in women. Survival probability has increased from 1940 to the present time. A high density of persons per square mile was associated with a higher-than-expected incidence. No trends in incidence according to socioeconomic status were observed.

Epidemiological characteristics of squamous cell carcinoma and adenocarcinoma of the bladder.

Recent incidence data from the United States indicate that transitional cell carcinoma accounts for the vast majority (95%) of bladder tumors in this country, with squamous cell carcinoma (less than 3%) and adenocarcinoma (less than 2%) comprising nearly all the remaining cases. Rates of squamous cell carcinoma and adenocarcinoma were higher in blacks compared to whites, while the reverse was true for transitional cell carcinoma. All three tumors predominated in males, especially transitional cell carcinoma. A population-based case-control study of bladder cancer conducted in 10 geographical areas of the United States identified 43 patients with squamous cell carcinoma and 32 with adenocarcinoma to permit an examination of risk factors. Cigarette smoking was significantly associated with risk of squamous cell carcinoma, with the relative risk rising to 6.1 among smokers of 40 or more cigarettes/day. Significantly elevated risks of squamous cell carcinoma were also associated with a history of 3 or more urinary tract infections (relative risk = 5.7) and with employment as welders and cooks. Risk factors were generally less conspicuous for adenocarcinoma, except for a significant trend with the amount of coffee drinking; however, this finding is based on small numbers and should be interpreted cautiously.

Follow-up study of twenty-four families with Li-Fraumeni syndrome.

The Li-Fraumeni cancer family syndrome is manifested by susceptibility to breast cancer, sarcomas, and other neoplasms in children and young adults. The present study utilized clinical follow-up data on 545 members of 24 Li-Fraumeni kindreds living and cancer-free at family ascertainment. Two hypotheses were tested based on a model of autosomal dominant genetic predisposition: (a) that syndrome cancers would continue to occur excessively during follow-up compared to the general population, and (b) that the tumors would occur primarily among those family members likely to carry the gene. Population cancer rates were compared with cancer rates in follow-up of the cohort from ascertainment to 1988. Risk of carrying the gene for the syndrome at the time of ascertainment was calculated for each family member under two models with somewhat different definitions of affection with the syndrome. Cancer occurrence after ascertainment was then analyzed according to the risks. Cancer did continue to occur excessively among the entire cohort during follow-up [relative risk (RR 2.1)]. The excess was greatest below age 20 (RR 21.1), declined with increasing age, and was most pronounced for neoplasms featured in the syndrome (RR 18.2). Among persons less than age 45, at least 87% of cancers occurred in those at higher risk of carrying the gene under both genetic models (RR 22.9 and 21.3). The clinical data, therefore, reliably identify individuals likely to carry a dominantly inherited gene conferring susceptibility to a specific constellation of neoplasms. Recent identification of a germ line mutation in the tumor suppressor gene p53 in persons with the syndrome may, if confirmed, have implications for ultimately defining the component tumors of the syndrome and for the causes and prevention of those tumors arising outside these families.

HLA antigens in renal cell carcinoma.

HLA antigen type was studied in 35 renal cell carcinoma patients who had bilateral disease, an early age of onset (less than age 45), or a family history of kidney cancer. Increased frequencies of the single-locus antigens HLA-DR8 (relative risk, 3.3) and HLA-Bw44 (relative risk, 2.1) and a deficit of HLA-DR1 (relative risk, 0.4) were found. Although based on small numbers, the relative excess was highest among persons phenotypically HLA-Bw44DR8. A higher frequency of the three-locus phenotype HLA-A3B7DR2 was also noted. The unusual HLA patterns were most pronounced among patients of German or Scandinavian origin, population groups reported to have an elevated risk of renal cancer.

Stainless steel surface functionalization for immobilization of antibody fragments for cardiovascular applications.

Stainless steel 316 L material is commonly used for the production of coronary and peripheral vessel stents. Effective biofunctionalization is a key to improving the performance and safety of the stents after implantation. This paper reports the method for the immobilization of recombinant antibody fragments (scFv) on stainless steel 316 L to facilitate human endothelial progenitor cell (EPC) growth and thus improve cell viability of the implanted stents for cardiovascular applications. The modification of stent surface was conducted in three steps. First the stent surface was coated with titania based coating to increase the density of hydroxyl groups for successful silanization. Then silanization with 3 aminopropyltriethoxysilane (APTS) was performed to provide the surface with amine groups which presence was verified using FTIR, XPS, and fluorescence microscopy. The maximum density of amine groups (4.8*10(-5) mol/cm(2)) on the surface was reached after reaction taking place in ethanol for 1 h at 60 °C and 0.04M APTS. On such prepared surface the glycosylated scFv were subsequently successfully immobilized. The influence of oxidation of scFv glycan moieties and the temperature on scFv coating were investigated. The fluorescence and confocal microscopy study indicated that the densest and most uniformly coated surface with scFv was obtained at 37 °C after oxidation of glycan chain. The results demonstrate that the scFv cannot be efficiently immobilized without prior aminosilanization of the surface. The effect of the chemical modification on the cell viability of EPC line 55.1 (HucPEC-55.1) was performed indicating that the modifications to the 316 L stainless steel are non-toxic to EPCs.

Location and dynamics of a membrane-bound fluorescent hapten. A spectroscopic study.

In the preceding paper (Petrossian, A. and Owicki, J.C. (1984), Biochim. Biophys. Acta 776, 217-227), we describe the binding of a monoclonal anti-fluorescein antibody to a membrane bound fluorescein-lipid hapten. Those results suggest that some of the hapten fluorescein moiety is extended away from the membrane surface and is available for antibody binding, while some of the hapten is sequestered and not immediately available for antibody binding. In this paper, we carry out a spectroscopic study of the membrane-bound hapten and show that there is more than one physically distinct fluorophore environment, with the sequestered hapten associated with the phospholipid headgroup region. The amount of membrane-associated fluorophore depends upon the membrane lipid composition: most of the fluorophore is associated when the lipid is unsaturated or branched-chain phosphatidylcholines (PC), whereas the hapten is largely extended for PC/cholesterol mixtures. The effect of cholesterol on the availability of membrane-bound hapten to antibody binding is not unique to this system. The conversion between sequestered and extended hapten is slow (minutes).

Hypertension in long-term survivors of childhood renal cancers.

The prevalence of hypertension was investigated in 119 adults who have survived for up to 53 years following the diagnosis of renal cancer in childhood (Wilms' tumor, 116 patients; renal carcinoma, three patients). Twenty-four (20%) have developed definite or borderline hypertension, as compared with 18.1 cases expected based on US population rates (relative risk [RR], 1.3; 95% confidence interval [CI], 0.9 to 2.0; P = .20). This nonsignificant excess is due to the heightened prevalence of definite hypertension among one subgroup of male patients. The findings are not explained by cigarette smoking, obesity, age, and stage at diagnosis of Wilms' tumor, or family history of hypertension. A case-comparison analysis within the cohort showed no consistent hypertensive effect associated with radiation therapy dose, radiotherapy concurrent with dactinomycin chemotherapy, or extent of renal surgery. Hypertension is not a common late complication of Wilms' tumor in our patients.

Cardiac Donor Risk Factors Predictive of Short-Term Heart Transplant Recipient Mortality: An Analysis of the United Network for Organ Sharing Database.

INTRODUCTION: To address the shortage of donor hearts for transplantation, there is significant interest in liberalizing donor acceptance criteria. Therefore, the aim of this study was to evaluate cardiac donor characteristics from the United Network for Organ Sharing (UNOS) database to determine their impact on posttransplantation recipient outcomes. METHODS: Adult (≥18 years) patients undergoing heart transplantation from July 1, 2004, to December 31, 2012, in the UNOS Standard Transplant Analysis and Research (STAR) database were reviewed. Patients were stratified by 1-year posttransplantation status; survivors (group S, n = 13,643) and patients who died or underwent cardiac retransplantation at 1-year follow-up (group NS/R = 1785). Thirty-three specific donor variables were collected for each recipient, and independent donor predictors of recipient death or retransplantation at 1 year were determined using multivariable logistic regression analysis. RESULTS: Overall 1-year survival for the entire cohort was 88.4%. Mean donor age was 31.5 ± 11.9 years, and 72% were male. On multivariable logistic regression analysis, donor age >40 years (odds ratio [OR] 1.44, 95% confidence interval [CI] 1.27 to 1.64), graft ischemic time >3 hours (OR 1.32, 1.16 to 1.51), and the use of cardioplegia (OR 1.17, 1.01 to 1.35) or Celsior (OR 1.21, 1.06 to 1.38) preservative solution were significant predictors of recipient death or retransplantation at 1 year posttransplantation. Male donor sex (OR 0.83, 0.74 to 0.93) and the use of antihypertensive agents (OR 0.88, 0.77 to 1.00) or insulin (OR 0.84, 0.76 to 0.94) were protective from adverse outcomes at 1 year. CONCLUSIONS: These data suggest that donors who are older, female, or have a long projected ischemic time pose greater risk to heart transplant recipients in the short term. Additionally, certain components of donor management protocols, including antihypertensive and insulin administration, may be protective to recipients.

Large-scale preparation and characterization of recombinant ovine placental lactogen.

To clone ovine placental lactogen (oPL) cDNA, total RNA from sheep placental cotyledon was reverse transcribed and the single-stranded cDNA was PCR-amplified with 5' and 3' primers containing, respectively, NcoI and PstI sites. The oPL cDNA fragment amplified between these two primers extended from A(-1) to the natural stop codon. The PCR product was gel-purified and subcloned into a Puc vector and the insert was sequenced on both strands, revealing several differences relative to the published sequence: S19N, S69N, D129E and R165Q. We assume that these differences can be accounted for by the high level of individual polymorphism, which has been described in detail for PLs of different species. The insert was subcloned into NcoI/ PstI-digested pTrc99A procaryotic expression plasmid and protein expression was induced by isopropyl-1-thio-beta-D-galactopyranoside. Because of low expression, oPL's cDNA was further subcloned into pET8 procaryotic expression plasmid. Its expression in E. coli strain BL21 transformed with this vector yielded 30-40 mg/l. The expressed protein, found in the inclusion bodies, was refolded into a monomer and purified on a Q-Sepharose column to homogeneity. Structural analysis using circular dichroism revealed a spectrum similar to that of human GH (hGH) thereby indicating proper refolding. Gel filtration and binding experiments, including real-time kinetic measurements using the surface plasmon resonance method revealed that oPL forms transient homodimeric complexes with extracellular domains of prolactin receptors from rabbit, rat and bovine and with hGH receptor. The purified oPL was biologically active in an Nb2-11C cell proliferation bioassay, in its ability to stimulate beta-casein synthesis in explants of ovine and rabbit mammary gland and fat synthesis in explants of bovine mammary gland, and in a proliferation assay using FDC-P1 cells transfected with rabbit or hGH receptors.

Tuberculosis chemotherapy and risk of bladder cancer.

In a population-based study of 2982 bladder cancer patients and 5782 population controls from 10 geographical areas of the US, no excess risk was associated with medications used for tuberculosis treatment or prophylaxis (relative risk (RR) = 0.95). The findings agree with other epidemiological studies that have not confirmed earlier reports linking isoniazid (INH) exposure to bladder cancer.

Characteristics and development of the murine B-1b (Ly-1 B sister) cell population.

In this paper we have outlined the evidence for two distinct branches of the B-1 cell lineage. The data show that phenotypically B-1a and B-1b cells are essentially identical, distinguished only by the presence or absence of the CD5 antigen. Functionally no differences between the two populations have yet been identified. Both produce anti-PtC antibodies, a specificity not observed in conventional B cells. Both produced high levels of IgM as measured in adoptive transfer experiments. Developmentally, B-1a and B-1b cells are indistinguishable with respect to generation from progenitors present in fetal liver and omentum, feedback regulation of new B-1a and B-1b cells from bone marrow, self-replenishment from Ig+ cells following adoptive transfer, and the generation of clonal populations. The major difference in the two populations is seen in the development of B-1a and B-1b cells from B220- progenitors in the adult bone marrow. Although B220- B-1a progenitors are rare in adult (greater than 6 weeks) bone marrow, the progenitors for B-1b cells persist well into adulthood. Our understanding of B-1b cell ontogeny is at a stage similar to that of B-1a cells five years ago. We have evidence from transfer experiments that strongly suggests the existence of two distinct progenitors for B-1a and B-1b, but we have yet to physically separate these progenitors as Solvansen et al. have done for B-1 and conventional B cells. Furthermore we must determine whether the B-1b cells that develop from fetal liver and bone marrow are functionally and developmentally equivalent to those that develop from adult bone marrow. As with B-1a cells, the role of B-1b cells in the immune system is unclear. Although we have not yet discerned functional differences between B-1a and B-1b, given the recent identification of CD72 (Lyb-2) as the ligand for CD5, it is tempting to speculate that B-1a cells are more involved in B-B cell interactions such as idiotype-anti-idiotype regulation of the early B-cell repertoire and that B-1b cells are more involved in B-T cell interactions. Whatever their function, it is clear that in trying to understand the role of the B-1 lineage it is important to consider both the B-1a and B-1b lineages.

Molecular characterization of the replicon of the Pediococcus pentosaceus 43200 pediocin A plasmid pMD136.

The pediocin A-encoding plasmid of Pediococcus pentosaceus 43200, pMD136, was characterized by restriction enzyme analysis. Analysis of its replicon was facilitated by the construction of a probe vector consisting of the Escherichia coli plasmid pSP72 and the cat gene from Staphylococcus aureus plasmid pC194. The replication region of pMD136 was localized on a 1.6-kb EcoRI/BglII fragment. Sequencing analysis revealed a non-coding region, repA, spanning the first 440 bp, followed by an open reading frame, repB, encoding a putative protein of 390 amino acids. The non-coding region contained two sets of 6-bp and two sets of 22-bp direct repeats and two sets of inverted repeats upstream of the open reading frame. Strong homology of the isolated replicon was found to theta-type replicons of Lactococcus lactis plasmids. Segregational stability assay suggested at least two regions as potentially involved in the stabilization of pMD136. The plasmid's strong homology to other theta-type replicons and its relatively high stability suggest that pMD136 belongs to the widespread family of theta-replication plasmids.

Occupations of fathers of patients with Wilms's tumour.

A case-control study of 149 Connecticut-born children with Wilms's tumour reported to the Connecticut Tumor Registry during the period 1935--1973 and of 149 matched controls was undertaken in order to explore the possibility that children with Wilms's tumour may have been exposed perinatally to carcinogenic agents. The occupation of the father at the time of the child's birth was investigated and used as an indicator of potential sources of carcinogens to which infants in the study may have been exposed. An association was found between paternal occupations related to lead in the group developing Wilms's tumour compared with the controls.

The role of interventional radiology in the management of genitourinary trauma.

The role of the radiologist in the management of urologic disease has changed dramatically in the last 10 years with the introduction of new imaging methods and the evolution of the angiographic catheter into a therapeutic instrument. The authors review the diagnostic options in genitourinary trauma, the indications for and techniques of transcatheter arterial embolization for renal and other retroperitoneal hemorrhage, and nonvascular interventions such as antegrade nephrostomy, stenting of urethral disruptions, percutaneous bladder drainage, and drainage of abscesses, hematomas, and urinomas.

Epidemic AIDS-related Kaposi's sarcoma in southern Africa: experience at the Johannesburg General Hospital (1980-1990).

Epidemic acquired immune deficiency syndrome-related Kaposi's sarcoma (AKS) in tropical and southern Africa is a highly varied neoplastic disease, characterized by multifocal mucocutaneous, lymphatic and visceral involvement. It follows a clinical course similar to AKS in Europe and the USA. However, lack of adequate medical facilities in many African countries hampers successful palliation of this fatal disease. In this retrospective analysis, we summarize our experience with 52 patients with AKS treated at Johannesburg General Hospital, South Africa, between 1980 and 1990. Radiation therapy can provide good to excellent palliation with only minimal side-effects, producing a lesser impact on the haematological and immunological system than chemotherapy.

PIP: Researchers analyzed data on 52 HIV-positive patients with Kaposi's sarcoma (KS) aged 23-67 (74% Black, 26% White; male/female ratio = 2.8:1) referred to the Johannesburg General Hospital in South Africa during 1980-1990 to examine the hospital's experience with these patients. 23 patients had a fever and/or at least 10% weight loss. 34% had prior or coexistent opportunistic infection, particularly Pneumocystis carinii pneumonia, fungal disease, or tuberculosis. Possible risk factors among 21 patients were homosexual intercourse, history of sexually transmitted disease, and drug abuse. Almost all patients had skin disease, either localized or disseminated. Other KS sites included the oral cavity, regional lymph nodes, and large bowel. 90% of 20 patients treated with radiation responded to treatment. Response rates for radiation treatment among the 20 patients were 80% for symptomatic relief, 45% for complete remission, 45% for partial remission, and 10% for tumor progression. The recurrence-free period among irradiated patients was five months. Five patients developed radiation-induced mucositis of the oropharyngeal region. None of the 32 patients treated with chemotherapy and not radiation experienced complete remission. Chemotherapy induced partial remission in 38% and tumor progression in 62% of patients. 9% of chemotherapy-treated patients experienced symptomatic relief. Deteriorating performance status and/or debilitating side effects (severe mucositis and neutropenic sepsis) necessitated cessation of chemotherapy or dose modification. The clinical course of AIDS-related KS in this population paralleled that in Western countries. Based on these findings, the authors recommend local radiation therapy to treat AIDS-related KS or a watch-and-wait policy for asymptomatic, minimal disease in patients with an intact immune status.

Evaluation of infectious crystalline keratitis with confocal microscopy in a case series.

We sought to determine whether there are unique findings in infections crystalline keratitis (ICK) examined by confocal microscopy and if confocal microscopy is predictive for bacteriology in ICK. A retrospective review of consecutive patients with a presumed diagnosis of ICK by slit-lamp examination was performed. These patients were then examined with confocal microscope and cultured. Sixteen patients were identified by biomicroscopy. Average age was 71 years; 12 of 16 patients were women; 10 of 16 had prior penetrating keratoplasty; and 12 of 16 were taking topical steroids. Confocal microscopy revealed a variable appearance to the crystals in the corneal stroma. Eight of 16 patients had distinct needle-like deposits at varying depths in the stroma, and eight had amorphous deposits grouped at different levels of the stroma. The results of confocal microscopic examination resembled the reported histopathology with clusters of deposits, but its current resolution does not allow identification of bacterial morphology. There was no correlation of morphology with culture results. Organisms were recovered in 12 of 16 patients by culture. In 10 of 16 patients, the infection was successfully treated with topical antibiotics, usually cefazolin. Crystal morphology of ICK can be observed by confocal microscopy. No pathognomonic, single pattern for this disease is seen with the confocal microscope. The latter may be an aid in determining the clinical response to treatment.

Radiation therapy in an elderly African population with Kaposi's sarcoma.

There are no published studies on the efficacy of modern radiation therapy in elderly populations with the endemic African type of Kaposi's sarcoma (AKS). The present retrospective analysis of 20 elderly AKS patients treated by radiotherapy attempts to supply information relevant to the older age group. It demonstrates that excellent symptomatic relief with minimal side effects can be attained and suggests that the role of radiotherapy as the treatment of choice in this particular group should be emphasized.

Radiation therapy in epidemic, AIDS-related Kaposi's sarcoma in southern Africa.

AIMS AND BACKGROUND: Acquired Immunodeficiency Syndrome (AIDS) associated Kaposi's Sarcoma (EKS) is widely spread in the Southern African Region. No large studies concerning the role of radiation therapy in the Southern African variant of EKS have been reported to date. METHODS: Over a 10 year period (1982-1992) 25 patients with EKS (disseminated skin involvement) were treated primarily with radiation therapy at the Johannesburg General Hospital. Radiation fields were individually tailored to the extent of the disease. Total administered doses ranged between 8-12 Gy (single fraction) to 24-30 Gy fractionated over 2-3 weeks. RESULTS: Overall response and symptomatic relief rates were 72% and 80%, respectively. Toxicity was mild and manageable. CONCLUSIONS: Our retrospective analysis supports the use of radiation therapy for the Southern African type of EKS.

PIP: Data suggest that 10-20% of African HIV-infected persons have Kaposi's Sarcoma (KS). African epidemic, AIDS-related KS (EKS) is widespread in the southern African region, with patients often needing treatment because of the disfiguring and stigmatic nature of the disease. Cytotoxic chemotherapy has shown antitumor activity, but it may further compromise the underlying immune deficiency. EKS is, however, very radiosensitive and radiation therapy is considered to be the treatment of choice for palliation, despite the absence of large studies concerning the role of radiation therapy in the southern African variant of EKS reported to date. The authors report findings from a 1982-92 study of radiation therapy among 25 patients with EKS at the Johannesburg General Hospital. Radiation fields were individually tailored to the extent of the disease. Total administered doses ranged 8-12 Gy (single fraction) to 24-30 Gy fractionated over 2-3 weeks to yield 72% and 80% overall response and symptomatic relief rates, respectively. Toxicity was mild and manageable. This retrospective analysis therefore supports the use of radiation therapy for the southern African type of EKS.