Implementation of the North Carolina HIV Bridge Counseling Program to Facilitate Linkage and Reengagement in Care for Individuals Infected with HIV/AIDS.

BACKGROUND Statewide interventions are critical to meeting the goals of the National HIV/AIDS Strategy in this country. In 2012, the North Carolina Division of Public Health developed the North Carolina State Bridge Counselor program to improve linkage to and reengagement in care for newly diagnosed persons and persons living with HIV who were out-of-care.METHODS We reviewed the planning process for the North Carolina State Bridge Counselor program, which involved a review of existing strengths-based counseling models for persons living with HIV, implementation of these models, and communication strategies with other providers. State bridge counselor responsibilities were delineated from the role of disease intervention specialists while retaining the fieldwork capability of disease intervention specialists to conduct outreach and provide services for persons living with HIV throughout the state.RESULTS Program implementation required extensive planning with stakeholders, incorporation of strengths-based counseling models, development of performance standards, and utilization of CAREWare, an HIV care software program to document referrals and data-sharing between state bridge counselors and clinics. By the end of 2014, state bridge counselor services were provided to approximately 60 of the 400 persons living with HIV (15%) who are diagnosed each quarter in North Carolina, with increasing utilization of the program.LIMITATIONS We assessed the development of this intervention specific to the North Carolina Division of Public Health, which may limit its generalizability. However, the State Bridge Counselor program was implemented in both urban and rural areas throughout the state, which increases its applicability to different public health programs throughout the country.CONCLUSION We demonstrated that a statewide State Bridge Counselor program for linkage and reengagement activities can be implemented by leveraging existing infrastructures, electronic medical records, HIV care networks, and fieldwork activities.

Prognostic value of pretreatment circulating neutrophils, monocytes, and lymphocytes on outcomes in lung stereotactic body radiotherapy.

PURPOSE: In the present study, we determined the association of pretreatment circulating neutrophils, monocytes, and lymphocytes with clinical outcomes after lung stereotactic body radiotherapy (sbrt). METHODS: All patients with primary lung cancer and with a complete blood count within 3 months of lung sbrt from 2005 to 2012 were included. Overall survival (os) was calculated using the Kaplan-Meier method. Factors associated with os were investigated using univariable and multivariable Cox proportional hazards regression. Fine-Gray competing risk regression was performed to test the association of the neutrophil:lymphocyte (nlr) and monocyte:lymphocyte (mlr) ratios with two types of failure: disease-related failure and death, and death unrelated to disease. RESULTS: Of the 299 sbrt patients identified, 122 were eligible for analysis. The median and range of the nlr and mlr were 3.0 (0.3-22.0) and 0.4 (0.1-1.9) respectively. On multivariable analysis, sex (p = 0.02), T stage (p = 0.04), and nlr (p < 0.01) were associated with os. On multivariable analysis, T stage (p < 0.01) and mlr (p < 0.01) were associated with disease-related failure; mlr (p = 0.03), nlr (p < 0.01), and sbrt dose of 48 Gy in 4 fractions (p = 0.03) and 54 Gy or 60 Gy in 3 fractions (p = 0.02) were associated with disease-unrelated death. Median survival was 4.3 years in the nlr≤3 group (95% confidence interval: 3.5 to not reached) and 2.5 years in the nlr>3 group (95% confidence interval: 1.7 to 4.8; p < 0.01). CONCLUSIONS: In lung sbrt patients, nlr and mlr are independently associated with os and disease-unrelated death. If validated, nlr and mlr could help to identify patients who would benefit most from sbrt.

The prevalence and nature of supportive care needs in lung cancer patients.

PURPOSE: In the present work, we set out to comprehensively describe the unmet supportive care and information needs of lung cancer patients. METHODS: This cross-sectional study used the Supportive Care Needs Survey Short Form 34 (34 items) and an informational needs survey (8 items). Patients with primary lung cancer in any phase of survivorship were included. Demographic data and treatment details were collected from the medical charts of participants. The unmet needs were determined overall and by domain. Univariable and multivariable regression analyses were performed to determine factors associated with greater unmet needs. RESULTS: From August 2013 to February 2014, 89 patients [44 (49%) men; median age: 71 years (range: 44-89 years)] were recruited. The mean number of unmet needs was 8 (range: 0-34), and 69 patients (78%) reported at least 1 unmet need. The need proportions by domain were 52% health system and information, 66% psychological, 58% physical, 24% patient care, and 20% sexuality. The top 2 unmet needs were "fears of the cancer spreading" [n = 44 of 84 (52%)] and "lack of energy/tiredness" [n = 42 of 88 (48%)]. On multivariable analysis, more advanced disease and higher MD Anderson Symptom Inventory scores were associated with increased unmet needs. Patients reported that the most desired information needs were those for information on managing symptoms such as fatigue (78%), shortness of breath (77%), and cough (63%). CONCLUSIONS: Unmet supportive care needs are common in lung cancer patients, with some patients experiencing a very high number of unmet needs. Further work is needed to develop resources to address those needs.

The association between socio-demographic marginalization and plasma glucose levels at diagnosis of gestational diabetes.

AIMS: We examined the association between socio-demographic marginalization and plasma glucose levels at diagnosis of gestational diabetes in a multi-ethnic and socio-economically diverse patient group. METHODS: Medical charts at a Toronto gestational diabetes clinic were reviewed for women with a recorded pregnancy between 1 March 2006 and 26 April 2011. One-hour 50-g glucose challenge test values and postal code data were abstracted. Postal codes were merged with 2006 Canadian census data to compute neighbourhood-level ethnic concentration (% recent immigrants, % visible minorities) and material deprivation (% low education, % low income, single-parent households). We compared women in the highest neighbourhood quintiles for both ethnic concentration and material deprivation with all other women to explore an association between marginalization and diagnostic glucose levels. Multivariate regression models of glucose challenge test values and insulin prescription were adjusted for age, prior gestational diabetes, parity and diabetes family history. RESULTS: Among 531 patients with complete glucose challenge test data (mean 11.94 mmol/l, sd 1.83), those in the most marginalized neighbourhoods had 0.43 mmol/l higher glucose challenge test values (95% CI 0.08-0.78) compared with the rest of the study population. Other factors associated with higher glucose challenge test values were prior gestational diabetes (0.59 mmol/l increment, 95% CI 0.19-0.99) and diabetes family history (0.32 mmol/l increment, 95% CI -0.01 to 0.66). Each additional 1 mmol/l glucose challenge test result was associated with an increased likelihood of being prescribed insulin (odds ratio 1.33, 95% CI 1.17-1.51). CONCLUSIONS: Women living in the most materially deprived and ethnically concentrated neighbourhoods have higher glucose levels at diagnosis of gestational diabetes. They may need close monitoring for timely initiation of insulin.

Novel therapeutic receptor agonists and antagonists in allergic conjunctivitis.

PURPOSE OF REVIEW: Allergic conjunctivitis is characterized by the development of pathophysiological changes to the ocular surface, which occurs when pro-allergic and pro-inflammatory mediators interact with their cognate receptors expressed on immune and nonimmune cells. Traditional treatments with antihistamines and corticosteroids provide relief, but there is a need for more efficacious and tolerable long-term therapy with a better safety profile. This article aims to provide an overview of the mode of action and clinical application of agonist therapies targeting glucocorticoid, melanocortin, and toll-like receptors, as well as antagonist therapies targeting cytokine, chemokine, integrin, and histamine receptors. RECENT FINDINGS: There has been considerable advancement in immunology and pharmacology, as well as a greater understanding of the cellular and molecular mechanisms of allergic conjunctivitis. Recent research advancing therapy for allergic conjunctivitis has focused on developing synthetic molecules and biologics that can interfere with the process of the allergic immune reaction. SUMMARY: This review discusses novel therapeutic receptors being explored agonistically or antagonistically to develop alternative treatment options for allergic conjunctivitis. These novel approaches hold promise for improving the management of allergic eye diseases, offering patients hope for more effective and safer treatment options in the future.

Cytokines in Allergic Conjunctivitis: Unraveling Their Pathophysiological Roles.

Allergic conjunctivitis is one of the common immune hypersensitivity disorders that affect the ocular system. The clinical manifestations of this condition exhibit variability contingent upon environmental factors, seasonal dynamics, and genetic predisposition. While our comprehension of the pathophysiological engagement of immune and nonimmune cells in the conjunctiva has progressed, the same cannot be asserted for the cytokines mediating this inflammatory cascade. In this review, we proffer a comprehensive description of interleukins 4 (IL-4), IL-5, IL-6, IL-9, IL-13, IL-25, IL-31, and IL-33, as well as thymic stromal lymphopoietin (TSLP), elucidating their pathophysiological roles in mediating the allergic immune responses on the ocular surface. Delving into the nuanced functions of these cytokines holds promise for the exploration of innovative therapeutic modalities aimed at managing allergic conjunctivitis.

The Usefulness of Assessing Glaucoma Progression With Postprocessed Visual Field Data.

Purpose: Data postprocessing with statistical techniques that are less sensitive to noise can be used to reduce variability in visual field (VF) series. We evaluated the detection of glaucoma progression with postprocessed VF data generated with the dynamic structure-function (DSF) model and MM-estimation robust regression (MRR). Method: The study included 118 glaucoma eyes with at least 15 visits selected from the Rotterdam dataset. The DSF and MRR models were each applied to observed mean deviation (MD) values from the first three visits (V1-3) to predict the MD at V4. MD at V5 was predicted with data from V1-4 and so on until the MD at V9 was predicted, creating two additional datasets: DSF-predicted and MRR-predicted. Simple linear regression was performed to assess progression at the ninth visit. Sensitivity was evaluated by adjusting for false-positive rates estimated from patients with stable glaucoma and by using longer follow-up series (12th and 15th visits) as a surrogate for progression. Results: For specificities of 80% to 100%, the DSF-predicted dataset had greater sensitivity than the observed and MRR-predicted dataset when positive rates were normalized with corresponding false-positive estimates. The DSF-predicted and observed datasets had similar sensitivity when the surrogate reference standard was applied. Conclusions: Without compromising specificity, the use of DSF-predicted measurements to identify progression resulted in a better or similar sensitivity compared to using existing VF data. Translational Relevance: The DSF model could be applied to postprocess existing visual field data, which could then be evaluated to identify patients at risk of progression.

Dietary glycemic index, glycemic load, insulin index, fiber and whole-grain intake in relation to risk of prostate cancer.

OBJECTIVE: Insulin may play a role in prostate cancer tumorigenesis. Postprandial blood glucose and insulin responses of foods depend importantly on the carbohydrate quality and quantity, represented by glycemic index (GI), glycemic load (GL), fiber and whole-grain content, but are also influenced by intake of protein and other characteristics. The recently developed insulin index (II) quantifies the postprandial insulin secretion, also taking into account these additional characteristics. METHODS: We investigated the association between dietary GI, GL, II, fiber, and whole grains and risk of total prostate cancer (n = 5,112) and subgroups of prostate cancer as defined by stage or grade in 49,934 male participants of the Health Professionals Follow-up Study. Multivariate adjusted hazard ratios (HR) and 95% confidence intervals (95% CI) were estimated using Cox proportional hazards regression. RESULTS: Dietary GI, GL, II, or fiber was not associated with risk of total or subgroups of prostate cancer. We observed a positive association between dietary intake of whole grains and total prostate cancer (HR highest versus lowest quintile 1.13, 95% CI 1.03-1.24), which was attenuated after restriction to PSA-screened participants (HR 1.03, 95% CI 0.91-1.17). CONCLUSIONS: These results suggest that long-term exposure to a diet with a high insulin response does not affect prostate cancer incidence.

Detecting Progression in Patients With Different Clinical Presentations of Primary Open-angle Glaucoma.

PRCIS: Glaucoma progression was more frequently identified by assessing retinal fiber layer thickness than by monitoring visual field (VF) loss for different baseline classifications in primary open-angle glaucoma. PURPOSE: The aim was to compare the detection of glaucoma progression by retinal nerve fiber layer thickness (RNFLT) and VF assessments for different baseline classifications of primary open-angle glaucoma. METHODS: This study included 194 eyes from 194 patients with a minimum of 9 follow-up visits selected from the Diagnostic Innovation in Glaucoma Study (DIGS) and the African Descent and Glaucoma Evaluation Study (ADAGES). Each eye was classified according to baseline clinical signs: ocular hypertension (n=39), glaucomatous optic neuropathy only (n=60), glaucomatous visual field loss only (GVF, n=39) and definite glaucoma (concurrent optic disc and VF defect, n=56). We assessed progression by performing simple linear regression on global and sectorial mean deviations values generated for RNFLT (RNFLT-MD) and VF data (VF-MD). The proportion of eyes identified as progressing (positive rate) by RNFLT-MD and by VF-MD were compared within each classification. RESULTS: Whereas both parameters performed similarly among glaucomatous optic neuropathy only and definite glaucoma eyes, the positive rate obtained with global RNFLT-MD was significantly greater compared with global VF-MD by 33.3% and 30.8% among ocular hypertension eyes and GVF eyes, respectively. This finding was consistent in the inferotemporal sector; however, similar positive rates were obtained for both parameters in the superotemporal sector. CONCLUSIONS: While both RNFLT and VF parameters showed comparable abilities to identify progression across the different classifications, RNFLT assessment may be better suited to monitor progression, particularly among patients with elevated intraocular pressure and those who present with only GVF defect at baseline.

Nutrition knowledge and dietary patterns in ophthalmic patients.

CLINICAL SIGNIFICANCE: Ophthalmic nutrition education programs and good dietary patterns are of considerable significance to ocular health outcomes. BACKGROUND: The study aimed at assessing the level of ophthalmic nutrition knowledge and to compare the dietary patterns between different ophthalmic patients. METHODS: Four hundred and ninety-two adults (mean age 54.7 ± 19.4-years) receiving care at three eye-care facilities in Ghana were surveyed: 171 had senile cataract, 162 had open-angle glaucoma, and 159 had healthy eyes. A modified food frequency questionnaire was administered to elicit ophthalmic nutrition knowledge and dietary intake information of participants over the last 30-days. The results were compared between the three groups. RESULTS: More than half of the participants (n = 267; 54.3 per cent) had received some form of education on food sources that are good for the eyes and vision, with healthy participants accounting for 50.6 per cent of this proportion. Television (55.1 per cent; 147/267), radio (49.1 per cent; 131/267) and the eye doctor (34.1 per cent; 91/267) were the most frequent sources of ophthalmic nutritional knowledge. Overall, less than one-quarter of the participants (n = 108; 22.2 per cent) were able to correctly identify one food source for at least four out of seven selected ophthalmic nutrients. Vitamin C was the most frequently consumed ophthalmic nutrient, whereas lycopene was the least taken across all groups. The average ophthalmic nutrients intake score for those with healthy eyes (59.16 [53.39-64.93]) was significantly greater than for both glaucoma patients (37.73 [32.01-43.44]) and cataract patients (34.81 [29.24-40.37]). CONCLUSION: In addition to poor ophthalmic nutrition awareness and knowledge, both cataract and glaucoma patients, compared to those with healthy eyes, consumed a lesser amount of nutrients considered vital for eye health and vision. Ophthalmic nutrition education is needed to improve eye-health outcomes.

Electrode displacement strain imaging of thermally-ablated liver tissue in an in vivo animal model.

PURPOSE: Percutaneous thermal ablation is increasingly being used to destroy hepatic tumors in situ. The success of ablative techniques is highly dependent on adequate ablation zone monitoring, and ultrasound-based strain imaging could become a convenient and cost-effective means to delineate ablation zone boundaries. This study investigates in vivo electrode displacement-based strain imaging for monitoring hepatic ablation procedures that are difficult to perform with conventional elastography. METHODS: a In our method, minute displacements (less than a millimeter) are applied to the unconstrained end of the ablation electrode, resulting in localized tissue deformation within the ablation zone that provides the mechanical stimuli required for strain imaging. This article presents electrode displacement strain images of radiofrequency ablation zones created in porcine liver in vivo (n = 13). RESULTS: Cross-sectional area measurements from strain images of these ablation zones were obtained using manual and automated segmentation. Area measurements from strain images were highly correlated with areas measured on histopathology images, quantitated using linear regression (R = 0.894, P < 0.001 and R = 0.828, P < 0.001, respectively). CONCLUSIONS: This study further demonstrates that electrode displacement elastography is capable of providing high-contrast images using widely available commercial ultrasound systems which may potentially be used to assess the extent of thermal ablation zones.

Managing uncertain recovery for patients nearing the end of life in hospital: a mixed-methods feasibility cluster randomised controlled trial of the AMBER care bundle.

BACKGROUND: The AMBER (Assessment, Management, Best Practice, Engagement, Recovery Uncertain) care bundle is a complex intervention used in UK hospitals to support patients with uncertain recovery. However, it has yet to be evaluated in a randomised controlled trial (RCT) to identify potential benefits or harms. The aim of this trial was to investigate the feasibility of a cluster RCT of the AMBER care bundle. METHODS: This is a prospective mixed-methods feasibility cluster RCT. Quantitative data collected from patients (or proxies if patients lack capacity) were used (i) to examine recruitment, retention and follow-up rates; (ii) to test data collection tools for the trial and determine their optimum timing; (iii) to test methods to identify the use of financial resources; and (iv) to explore the acceptability of study procedures for health professionals and patients. Descriptive statistical analyses and thematic analysis used the framework approach. RESULTS: In total, 894 patients were screened, of whom 220 were eligible and 19 of those eligible (8.6%) declined to participate. Recruitment to the control arm was challenging. Of the 728 patients screened for that arm, 647 (88.9%) were excluded. Overall, 65 patients were recruited (81.3% of the recruitment target of 80). Overall, many were elderly (≥80 years, 46.2%, n = 30, mean = 77.8 years, standard deviation [SD] = 12.3 years). Over half (53.8%) had a non-cancer diagnosis, with a mean of 2.3 co-morbidities; 24.6% patients (n = 16) died during their hospital stay and 35.4% (n = 23) within 100 days of discharge. In both trial arms, baseline IPOS subscale scores identified moderate patient anxiety (control: mean 13.3, SD 4.8; intervention: mean 13.3, SD 5.1), and howRwe identified a good care experience (control: mean 13.1, SD 2.5; intervention: mean 11.5, SD 2.1). Collecting quantitative service use and quality of life data was feasible. No patient participants regarded study involvement negatively. Focus groups with health professionals identified concerns regarding (i) the subjectivity of the intervention's eligibility criteria, (ii) the need to prognosticate to identify potential patients and (iii) consent procedures and the length of the questionnaire. CONCLUSIONS: A full trial of the AMBER care bundle is technically feasible but impractical due to fundamental issues in operationalising the intervention's eligibility criteria, which prevents optimal recruitment. Since this complex intervention continues to be used in clinical care and advocated in policy, alternative research approaches must be considered and tested. TRIAL REGISTRATION: International Standard Randomised Controlled Trial Number (ISRCTN) Register, ISRCTN36040085 .

Associations between neighborhood-level violence and individual mental disorders: Results from the World Mental Health surveys in five Latin American cities.

Rapidly urbanizing areas of Latin America experience elevated but unevenly distributed levels of violence. Extensive research suggests that individual exposure to violence is associated with higher odds of both internalizing (anxiety and mood) and externalizing (substance and intermittent explosive) mental disorders. Less research, however, has focused on how neighborhood-level violence, as an indicator of broader neighborhood contexts, might relate to the mental health of residents, independently of an individual's personal exposure. We used multilevel analyses to examine associations of neighborhood-level violence with individual-level past-year mental disorders, controlling for individual-level violence exposure. We used data from 7,251 adults nested in 83 neighborhoods within five large Latin American cities as part of the WHO World Mental Health Surveys. Accounting for individual-level violence exposure, living in neighborhoods with more violence was associated with significantly elevated odds of individual-level internalizing disorders, but not externalizing disorders. Caution should be exercised when making causal inferences regarding the effects of neighborhood-level violence in the absence of experimental interventions. Nevertheless, neighborhood context, including violence, should be considered in the study of mental disorders. These findings are particularly relevant for rapidly urbanizing areas with high levels of violence, such as Latin America.

A study of human leukocyte D locus related antigens in Graves' disease.

An association between Graves' disease and the human leukocyte antigen (HLA) system has previously been reported. The disease was more strongly associated with the HLA D locus antigen Dw3 than with HLA B8. Products of the HLA D locus are determined by the interaction of test cells with standard typing lymphocytes, a technically difficult procedure. Recently, it has been possible to type serologically for D locus related (DRw) specificities on peripheral bone marrow-derived (B) lymphocytes. Blood B lymphocytes from 50 unrelated controls and 41 patients with Graves' disease were typed for seven HLA DRw specificities. 28 patients with Graves' disease (68%) were positive for DRw3, in contrast to 14 controls (28%); whereas only 21 patients (50%) were HLA B8 positive, compared with 13 (26%) controls. Thus, positivity for DRw3 afforded a relative risk for Graves' disease of 5.5, whereas that for HLA B8 amounted to 3.0. Additionally, a family with multiple cases of Graves' disease in which the disease was previously shown to be inherited with the haplotype, was linked to DRw2, which suggests that the susceptibility to the disease was inherited in association with that antigen. Two HLA B/glyoxalase recombination events were observed in this family; in both instances HLA DRw followed HLA B. This study thus demonstrates that the disease susceptibility gene for Graves' disease is in strong linkage disequilibrium with DRw3; however, it may be associated with other DRw specificities and inherited within family units in association with them.

Near vision spectacle coverage and barriers to near vision correction among adults in the Cape Coast Metropolis of Ghana.

PURPOSE: To determine the near vision spectacle coverage and barriers to obtaining near vision correction among adults aged 35 years and older in the Cape Coast Metropolis of Ghana. METHODS: A population-based cross-sectional study design was adopted and 500 out of 576 participants aged 35 years and older were examined from 12 randomly selected clusters in Cape Coast, Ghana. All participants underwent a comprehensive eye examination which included: distance and near visual acuities measurements and external and internal ocular health assessments. Distance and near refractions were performed using subjective refraction technique. Information on participants' demographics, near vision correction status, near visual needs and barriers to acquiring near vision correction were obtained through a questionnaire administered as part of the study. RESULTS: The mean age of participants was 52.3±10.3 years of whom 280 (56%) were females and 220 (44%) were males. The near vision spectacle coverage was 25%, 33% "met need" for near vision correction in the presbyopic population, and 64% unmet need in the entire study population. After controlling for other variables, age (5th and 6th decades) and educational level were associated with "met need" for near vision correction (OR=2.7 (1.55-4.68), p =0.00, and OR=2.36 (1.18-4.72), p=0.02 respectively). Among those who needed but did not have near vision correction, 64 (26%) did not feel the need for correction, 55 (22%) stated that they were unaware of available interventions, and 53 (21%) found the cost of near vision correction prohibitive. CONCLUSION: There was a low near vision spectacle coverage in this population which suggests the need for strategies on health education and promotion to address the lack of awareness of spectacle need and cost of services.

Vitamin A supplementation and plasma retinol levels: a randomized trial among women.

Although dietary intake of vitamin A has little, if any, overall effect on blood retinol in generally well-nourished populations, subgroups may exist that would be responsive to supplementation. The hypothesis that vitamin A supplementation increases blood retinol in apparently well-fed individuals with lower than usual blood levels was tested in female health workers, with relatively low blood retinol values, who were randomly assigned to receive vitamin A (10,000 IU daily) or placebo. After 4 weeks the mean change in plasma retinol was -0.4 micrograms/dl for the group receiving placebo and +4.1 micrograms/dl (an increase of 9% over base-line values) for the group receiving vitamin A (P = .02). The results were similar when the base-line retinol level and several other covariates were considered. Thirteen women who had initially received placebo were then switched to vitamin A for 4 weeks. These women experienced a mean increase of 5.3 micrograms/dl in plasma retinol (P = .04). Responses to vitamin A supplementation tend to be greater among women with lower previous total vitamin A intake, as assessed by questionnaire [Spearman rank correlation coefficient (r) = 0.50; P = .01].

Tamoxifen-mediated growth inhibition of human cholangiocarcinoma.

Cholangiocarcinoma represents a challenging primary malignancy of the liver with no effective medical therapy and a poor prognosis. We have investigated the role of tamoxifen and estrogen receptors (ERs) in the regulation of growth of human cholangiocarcinoma. Two human cholangiocarcinoma cell lines, OZ and SK-ChA-1, were grown in the presence of graded concentrations of tamoxifen; the effects on cell growth were determined by cell counting or 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium proliferation assay. The presence of ER protein was tested by indirect immunofluorescence and immunoprecipitation. In addition, cells were grown in estrogen-depleted media supplemented with exogenous 17beta-estradiol. ER mRNA was evaluated by reverse transcription-PCR and Northern blotting. Finally, one cholangiocarcinoma cell line was grown as a xenograft in athymic nude mice; tamoxifen effects on in vivo tumor growth were determined with biweekly caliper measurements. Tamoxifen (5-10 microM) caused dose-dependent in vitro growth inhibition of two human cholangiocarcinoma cell lines. In addition, growth inhibition of one cell line (SK-ChA-1) grown as a xenograft in nude mice by tamoxifen was observed. The presence of ER protein was suggested by 17beta-estradiol stimulation of tumor cell growth in vitro and confirmed by immunoprecipitation. Immunofluorescence microscopy was ineffective at detection of ER protein. Reverse transcription-PCR demonstrated the presence of ER mRNA in both cell lines. Northern blot analysis confirmed the presence of full-length 6.5-kb ER mRNA. No ER deletion mutants were detected. Tamoxifen inhibited the growth of human cholangiocarcinoma in vitro and in vivo. ER protein and mRNA were detected in both cell lines. The mechanism(s) of tamoxifen-mediated growth inhibition is unclear but may occur via ER protein or additional pathways. The ability of tamoxifen to inhibit tumor growth may offer an alternative adjunctive treatment for cholangiocarcinoma.

HLA-D--related (DRw) antigens in juvenile diabetes mellitus.

We studied the distribution of HLA-D--related (DRw) antigens in 40 patients with juvenile diabetes mellitus (JDM) and 79 matched controls. We found that DRw2 was significantly decreased in the JDM group, suggesting a protective effect of the antigen and that the decrease observed in B7 was secondary. HLA-DRw3 and HLA-DRw4 were increased in the diabetic group, and, as with B8/B15, these two antigen predisposed to the disease additively. The susceptibility for JDM was found to be more strongly related to HLA-DRw3 that to B8. On the other hand, B15 rather than DRw4 showed the stronger association with JDM. Moreover, we found that this second diabetogenic gene is associated primarily with B15 and only secondarily with Cw3, which is in linkage disequilibrium with B15. This study further emphasizes the immunogenetic heterogeneity of JDM.

Bacteriophage P22 portal protein is part of the gauge that regulates packing density of intravirion DNA.

The complex double-stranded DNA bacteriophages assemble DNA-free protein shells (procapsids) that subsequently package DNA. In the case of several double-stranded DNA bacteriophages, including P22, packaging is associated with cutting of DNA from the concatemeric molecule that results from replication. The mature intravirion P22 DNA has both non-unique (circularly permuted) ends and a length that is determined by the procapsid. In all known cases, procapsids consist of an outer coat protein, an interior scaffolding protein that assists in the assembly of the coat protein shell, and a ring of 12 identical portal protein subunits through which the DNA is presumed to enter the procapsid. To investigate the role of the portal protein in cutting permuted DNA from concatemers, we have characterized P22 portal protein mutants. The effects of several single amino acid changes in the P22 portal protein on the length of the DNA packaged, the density to which DNA is condensed within the virion, and the outer radius of the capsid have been determined. The results obtained with one mutant (NT5/1a) indicate no change (+/- 0.5%) in the radius of the capsid, but mature DNA that is 4.7% longer and a packing density that is commensurately higher than those of wild-type P22. Thus, the portal protein is part of the gauge that regulates the length and packaging density of DNA in bacteriophage P22. We argue that these findings make models for DNA packaging less likely in which the packing density is a property solely of the coat protein shell or of the DNA itself.

Molecular genetic analysis of bacteriophage P22 gene 3 product, a protein involved in the initiation of headful DNA packaging.

Bacteriophage P22 DNA packaging events occur in processive series on concatemeric phage DNA molecules. At the point where such series initiate, the DNA is recognized at a site called pac, and most molecular left ends are generated within six short regions called end sites, which are present in a 120 base-pair region surrounding the pac site. The bacteriophage P22 genes 2 and 3 proteins are required for successful generation of these ends and DNA packaging during progeny virion assembly. Mutants lacking the 162-amino-acid gene 3 protein replicate DNA and assemble functional procapsids. In this report we describe the nucleotide changes and DNA packaging phenotypes of a number of missense mutations of gene 3, which give the phage a higher than normal frequency of generalized transduction. In cells infected by these mutants, more packaging events initiate on the host chromosome than in wild-type infections, so the mutations are thought to affect the specificity of packaging initiation. In addition to having this phenotype, these mutations affect the process of phage DNA packaging in detectable ways. They may: (1) alter the target site specificity for packaging; (2) make target site recognition more promiscuous; (3) affect end site utilization; (4) alter the pac site; and (5) cause apparent random DNA packaging series initiation on phage DNA.