In-depth analysis of immunohistochemistry concordance in biopsy-resection pairs of bronchial carcinoids.

Primary diagnosis of bronchial carcinoids (BC) is always made on biopsies and additional immunohistochemistry (IHC) is often necessary. In the present study we investigated the concordance of common diagnostic (synaptophysin, chromogranin, CD56 and INSM-1) and potential prognostic (OTP, CD44, Rb and p16) IHC markers between the preoperative biopsies and resections of in total 64 BCs, 26 typical (41 %) and 38 atypical (59 %) carcinoid tumors. Synaptophysin and chromogranin had 100 % concordance in all resected carcinoids and paired diagnostic biopsies. Synaptophysin was not affected by variable expression in biopsies compared to chromogranin, CD56 and INSM-1. Notably, INSM-1 IHC was false negative in 8 % of biopsies. Of the novel and potential prognostic markers, only CD44 showed 100 % concordance between biopsies and resections, while OTP showed two (4 %) false negative results in paired biopsies. While Rb IHC was false negative in 8 % of biopsies, no strong and diffuse pattern of p16 expression was observed. In this study, most false negative IHC results (85 %, 22/26) were observed in small flexible biopsies. Taken together, our data suggest excellent concordance of synaptophysin and CD44 on the preoperative biopsy samples, while other neuroendocrine markers, Rb and OTP should be interpreted with caution, especially in small biopsies.

A Multimodal Biomarker Predicts Dissemination of Bronchial Carcinoid.

Background: Curatively treated bronchial carcinoid tumors have a relatively low metastatic potential. Gradation into typical (TC) and atypical carcinoid (AC) is limited in terms of prognostic value, resulting in yearly follow-up of all patients. We examined the additional prognostic value of novel immunohistochemical (IHC) markers to current gradation of carcinoids. Methods: A retrospective single-institution cohort study was performed on 171 patients with pathologically diagnosed bronchial carcinoid (median follow-up: 66 months). The risk of developing distant metastases based on histopathological characteristics (Ki-67, p16, Rb, OTP, CD44, and tumor diameter) was evaluated using multivariate regression analysis and the Kaplan−Meier method. Results: Of 171 patients, seven (4%) had disseminated disease at presentation, and 164 (96%) received curative-intent treatment with either endobronchial treatment (EBT) (n = 61, 36%) or surgery (n = 103, 60%). Among the 164 patients, 13 developed metastases at follow-up of 81 months (IQR 45−162). Univariate analysis showed that Ki-67, mitotic index, OTP, CD44, and tumor diameter were associated with development of distant metastases. Multivariate analysis showed that mitotic count, Ki-67, and OTP were independent risk factors for development of distant metastases. Using a 5% cutoff for Ki-67, Kaplan−Meier analysis showed that the risk of distant metastasis development was significantly associated with the number of risk predictors (AC, Ki-67 ≥ 5%, and loss of OTP or CD44) (p < 0.0001). Six out of seven patients (86%) with all three positive risk factors developed distant metastasis. Conclusions: Mitotic count, proliferation index, and OTP IHC were independent predictors of dissemination at follow-up. In addition to the widely used carcinoid classification, a comprehensive analysis of histopathological variables including Ki-67, OTP, and CD44 could assist in the determination of distant metastasis risks of bronchial carcinoids.

Diagnosis of atypical carcinoid can be made on biopsies > 4 mm2 and is accurate.

In the 2021 WHO thoracic tumors, gradation of lung carcinoids in biopsies is discouraged. We hypothesized that atypical carcinoid (AC) could be reliably diagnosed in larger preoperative biopsies. Biopsy-resection paired specimens of carcinoid patients were included, and definitive diagnosis was based on the resection specimen according to the WHO 2021 classification. A total of 64 biopsy-resection pairs (26 typical carcinoid (TC) (41%) and 38 AC (59%)) were analyzed. In 35 patients (55%), tumor classification between the biopsy and resection specimen was concordant (26 TC, 9 AC). The discordance in the remaining 29 biopsies (45%, 29 TC, 0 AC) was caused by misclassification of AC as TC. In biopsies measuring < 4 mm2, 15/15 AC (100%) were misclassified compared to 14/23 AC (61%) of biopsies ≥ 4 mm2. Categorical concordance of Ki-67 in biopsy-resection pairs at threshold of 5% was 68%. Ki-67 in the biopsy was not of additional value to discriminate between TC and AC, irrespective of the biopsy size. Atypical carcinoid is frequently missed in small bronchial biopsies (< 4 mm2). If the carcinoid classification is clinically relevant, a cumulative biopsy size of at least 4 mm2 should be considered. Our study provides strong arguments to make the diagnosis of AC in case of sufficient mitosis for AC on a biopsy and keep the diagnosis "carcinoid NOS" for carcinoids with ≤ 1 mitosis per 2 mm2. Ki-67 has a good concordance but was not discriminative for definitive diagnosis.

Sentinel lymph node biopsy for cutaneous melanoma: results of 10 years' experience in two regional training hospitals in the Netherlands.

BACKGROUND AND OBJECTIVE: The Multicenter Selective Lymphadenectomy Trial (MSLT-I) demonstrated that the sentinel node (SN) status in cutaneous melanoma affects prognosis and that completion lymphadenectomy in SN-positive patients may improve survival. Our objective was to evaluate sentinel lymph node biopsy (SLNB) in two regional hospitals in the Netherlands. METHODS: Patients with localized melanoma were planned for wide excision and SLNB. Completion lymphadenectomy was recommended for positive SN status. Data were compared with the MSLT-I. RESULTS: A median of 2 (1-7) SNs were identified in 305 patients and complications occurred in 11%. Fifty-four patients (18%) demonstrated SN metastases and 45 underwent completion lymphadenectomy (20% additional metastases). Six patients with initially negative SN developed lymph node metastases (sensitivity 90%). Overall disease-free survival was 83% (SN-negative 91% vs. SN-positive 41%; p < 0.001) and melanoma-specific survival was 93% (SN-negative 97% vs. SN-positive 62%; p < 0.001). Multivariate regression analysis revealed the SN status to be the most significant predictor for recurrence and melanoma-related death. CONCLUSION: Our results of SLNB are comparable to data from high-volume centers participating in MSLT-I. From a patient perspective, the false-negative SN rate of 10% and complication rate of 11% should be weighed against being informed about prognosis and having a possible therapeutic benefit from completion lymphadenectomy.

Abrogation of BRAFV600E-induced senescence by PI3K pathway activation contributes to melanomagenesis.

Human melanocytic nevi (moles) are benign lesions harboring activated oncogenes, including BRAF. Although this oncogene initially acts mitogenically, eventually, oncogene-induced senescence (OIS) ensues. Nevi can infrequently progress to melanomas, but the mechanistic relationship with OIS is unclear. We show here that PTEN depletion abrogates BRAF(V600E)-induced senescence in human fibroblasts and melanocytes. Correspondingly, in established murine BRAF(V600E)-driven nevi, acute shRNA-mediated depletion of PTEN prompted tumor progression. Furthermore, genetic analysis of laser-guided microdissected human contiguous nevus-melanoma specimens recurrently revealed identical mutations in BRAF or NRAS in adjacent benign and malignant melanocytes. The PI3K pathway was often activated through either decreased PTEN or increased AKT3 expression in melanomas relative to their adjacent nevi. Pharmacologic PI3K inhibition in melanoma cells suppressed proliferation and induced the senescence-associated tumor suppressor p15(INK4B). This treatment also eliminated subpopulations resistant to targeted BRAF(V600E) inhibition. Our findings suggest that a significant proportion of melanomas arise from nevi. Furthermore, these results demonstrate that PI3K pathway activation serves as a rate-limiting event in this setting, acting at least in part by abrogating OIS. The reactivation of senescence features and elimination of cells refractory to BRAF(V600E) inhibition by PI3K inhibition warrants further investigation into the therapeutic potential of simultaneously targeting these pathways in melanoma.

Effect of gender and nutritional status on academic achievement and cognitive function among primary school children in a rural district in Malaysia.

INTRODUCTION: The aim of this study was to investigate the relationship between gender, birth weight, nutritional status, and iron status of children with their academic performance and cognitive function. METHODS: Two hundred and forty-nine children, seven to nine years of age, were recruited by systematic sampling from six primary schools in a rural area in Malaysia. Cognitive function was assessed by using Raven's Coloured Progressive Matrices (R-CPM). Academic performance of the children was recorded from their school final examination results in four subjects including Malay language, English, Mathematics, and Science. Birth weight was recorded from the birth certificate, and nutritional status was determined by weight-for-age z score and height-for-age z score. RESULTS: Girls had a significantly higher score in all the academic tests, but a lower cognitive score compared to boys. Nutritional status was found to be correlated significantly with academic performance. Academic and cognitive function scores were also found to be correlated significantly with birth weight, parents' education, and family income. In a multivariate analysis, gender remained the significant predictor of academic function, and iron status and haemoglobin were the significant predictors of cognitive function, after controlling for other variables. CONCLUSION: The study showed that girls performed better academically than boys in rural Malaysia. Nutritional status, parents' education and family income could be additional modifiable factors to improve academic performance of the children. More attention is needed to improve academic achievements of boys at their early school years.

Preventing depression in high-risk groups.

PURPOSE OF REVIEW: Although most would agree that to prevent is better than to cure, prevention of depression has only recently been studied rigorously. The purpose of the present study is to review the state of the current literature. RECENT FINDINGS: The technical and theoretical literature underpinning depression prevention is developing in concert with high-quality intervention studies testing the effects of novel preventive interventions. Data suggest that universal prevention, targeting the whole population, is not likely to be effective, whereas both selective (high-risk groups) and indicated (people with some signs or symptoms, but no disorder) prevention may be very effective. Overall, preventive interventions may reduce the onset of depression by as much as 25-50%, which compares favourably with treatment. SUMMARY: Preventing depression may be effective at all ages and in diverse settings. Prevention has moved beyond the stage of pioneering studies and it deserves a regular place within our armamentarium to combat depression.

A Lethal Case of Fat Embolism Syndrome in a Nine-Year-Old Child: Options for Prevention.

We report a lethal case of fat embolism syndrome in a nine-year-old child after a direct blunt trauma leading to a pelvic fracture. On the second day, signs of bowel perforation and septic shock led to an acute aggravation of the pulmonary symptoms, cardiac arrest and death. Fat embolism is seldom thought to occur in pediatric trauma patients; however, this case illustrates it can lead to disastrous sequela. Since there is no specific treatment for it, prevention by early fracture stabilization is the only option.

Multivalent avimer proteins evolved by exon shuffling of a family of human receptor domains.

We have developed a class of binding proteins, called avimers, to overcome the limitations of antibodies and other immunoglobulin-based therapeutic proteins. Avimers are evolved from a large family of human extracellular receptor domains by in vitro exon shuffling and phage display, generating multidomain proteins with binding and inhibitory properties. Linking multiple independent binding domains creates avidity and results in improved affinity and specificity compared with conventional single-epitope binding proteins. Other potential advantages over immunoglobulin domains include simple and efficient production of multitarget-specific molecules in Escherichia coli, improved thermostability and resistance to proteases. Avimers with sub-nM affinities were obtained against five targets. An avimer that inhibits interleukin 6 with 0.8 pM IC50 in cell-based assays is biologically active in two animal models.

Activated leukocyte cell adhesion molecule (ALCAM/CD166/MEMD), a novel actor in invasive growth, controls matrix metalloproteinase activity.

Activated leukocyte cell adhesion molecule (ALCAM/CD166/MEMD) could function as a cell surface sensor for cell density, controlling the transition between local cell proliferation and tissue invasion in melanoma progression. We have tested the hypothesis that progressive cell clustering controls the proteolytic cascade for activation of gelatinase A/matrix metalloproteinase-2 (MMP-2), which involves formation of an intermediate ternary complex of membrane type 1 MMP (MT1-MMP/MMP-14), tissue inhibitor of metalloproteinase-2 (TIMP-2), and pro-MMP-2 at the cell surface. Surprisingly, truncation of ALCAM severely impaired MMP-2 activation in a nude mouse xenograft model, in which we previously observed diminished primary tumor growth and enhanced melanoma metastasis. Comparative studies of two-dimensional monolayer and three-dimensional collagen-gel cultures revealed that extensive cell-to-cell contacts, wild-type ALCAM, and cell-to-matrix interactions were all indispensable for efficient conversion of pro-MMP-2 to its active form in metastatic melanoma cells. Truncated, dominant-negative ALCAM diminished MMP-2 activation via reduced transcript levels and decreased processing of MT1-MMP. Failure of the proteolytic cascade after selective ALCAM depletion by RNA interference was mainly due to incomplete MT1-MMP processing, which was otherwise promoted by extensive cell-to-cell contacts. These data attribute a novel signaling role to ALCAM in regulation of proteolysis and support its previously postulated sensor function in invasive growth.

Activated leukocyte cell adhesion molecule (ALCAM/CD166): signaling at the divide of melanoma cell clustering and cell migration?

Orchestrated modulation of cell adhesion is essential for development and homeostasis in multicellular organisms. It optimizes embedding of the cell in its dynamic environment and facilitates appropriate cell responses and intercellular communication. Chronic disturbance of this delicate equilibrium causes defects in tissue architecture and sometimes cancer. In tumor cell biology, dynamic control of adhesion molecules is important to proceed through the metastatic cascade and to allow cell release from the primary tumor, invasion of the surrounding matrix, intravasation and adhesion to vascular endothelial cells to facilitate extravasation. Intertwined and multiple adhesive interactions rather than individual interactions presumably play critical roles in neoplastic development. Yet, knowledge of the contribution of each individual adhesion molecule is essential to unravel this network of interactions. This review will focus on activated leukocyte cell adhesion molecule (ALCAM/CD166) and its role in human melanoma progression. It is hypothesized that ALCAM may function as a cell surface sensor to register local growth saturation and to regulate cellular signaling and dynamic responses.

Direct binding of plectin to Fer kinase and negative regulation of its catalytic activity.

Plectin is a cyoskeletal linker protein that protects tissues against mechanical stress. We report here that the N-terminal domain of the nonreceptor tyrosine kinase Fer interacts with N-terminal sequences of plectin. Recombinant protein encoded by exon 12-24 of rat plectin bound directly to amino acid 1-329 of murine Fer. Using an antiserum prepared to a recombinant N-terminal fragment of Fer kinase, plectin was coimmunoprecipitated with Fer from cell lysates of cultured mouse fibroblasts. Plectin was shown to partially colocalize with Fer in these cells. Upon transfection of full length Fer cDNA into plectin-negative mouse fibroblasts, hyperphosphorylation of Fer was observed; hyperphosphorylation was strongly reduced when N-terminal Fer deletion mutants were transfected. Immunocomplex kinase assays showed that the activity of Fer kinase transfected into plectin-negative fibroblasts was increased compared to that transfected into wild type cells. We conclude that Fer interacts with plectin and that this interaction may serve to negatively regulate Fer's activity.

HIV prevalence remains low among Calgary's needle exchange program participants.

OBJECTIVES: To determine the demographic characteristics, risk behaviours and prevalence of HIV-1 among injection drug users (IDU) attending Calgary's needle exchange program (NEP). METHODS: A survey was conducted from June through September 1998 among IDU attending Calgary's NEP. Demographic and behaviour characteristics were determined by personal interview and saliva was tested for HIV antibody. RESULTS: There were 278 participants providing 272 saliva specimens. Nine were positive for HIV-1 (3.3%, 95% C.I. 1.6-6.4%). Sexual and injecting practices, cities where drugs had been used, incarceration, addiction treatment and demographic characteristics were described, and a subanalysis for women, youth and Aboriginals was carried out. CONCLUSIONS: HIV prevalence remains low among NEP attenders in Calgary, although high-risk behaviours are common. Women, youth and Aboriginals have unique risk behaviour profiles. Many IDU want to participate in addiction treatment, and strategies should be made to provide accessible, appropriate treatment services.

Adult formulation of a five component acellular pertussis vaccine combined with diphtheria and tetanus toxoids and inactivated poliovirus vaccine is safe and immunogenic in adolescents and adults.

BACKGROUND: Pertussis is increasingly recognized as an important cause of cough illness in adolescents and adults. PURPOSE: To evaluate the safety and antibody response to a single dose of an adult formulation of a five component (pertussis toxoid, filamentous hemagglutinin, pertactin, fimbriae 2 and 3) acellular pertussis vaccine (aP) combined with diphtheria and tetanus toxoids (TdaP) and inactivated poliovirus vaccine (TdaP-IPV) in adolescents and adults and to assess the response to a second dose of the acellular pertussis vaccine in a subset of the adults. POPULATION AND SETTING: The study addressed 1207 healthy participants (736 adults and 466 adolescents) recruited in five Canadian communities. STUDY DESIGN: In a randomized, observer-blind, controlled clinical trial, adult participants received Td followed at a separate visit by aP, TdaP followed by IPV or TdaP-IPV; adolescents received Td-IPV followed at a separate visit by aP or TdaP-IPV. A subgroup of adults was given a booster of aP 1 month after TdaP. OUTCOME MEASURES: Antibody titers measured before and 1 month after each immunization; adverse events enumerated at 24 h, 72 h and 8 to 10 days. RESULTS: The aP vaccine given by itself was associated with adverse events less frequently than were Td, Td-IPV, TdaP or TdaP-IPV vaccines, but reaction rates did not differ significantly among the latter products. The antibody response against Bordetella pertussis antigens was vigorous in all groups, although adults given the TdaP-IPV vaccine had lower antibody titers against filamentous hemagglutinin, pertactin, diphtheria and tetanus antibodies than those given TdaP vaccine. Similarly adolescents given TdaP-IPV had lower antibody titers against pertussis toxin, filamentous hemagglutinin, fimbriae and agglutinins than those given Td-IPV and aP alone. A second dose of acellular pertussis vaccine was not associated with increased adverse events in adults but elicited increased antibody titers over that achieved by a single dose only against pertussis toxin. CONCLUSIONS: This adult formulation five component aP vaccine given as TdaP-IPV is safe and immunogenic in adolescents and adults and is a candidate vaccine for adolescent and adult immunization programs.

Comparison of a fifth dose of a five-component acellular or a whole cell pertussis vaccine in children four to six years of age.

BACKGROUND AND OBJECTIVES: Acellular pertussis vaccines are now preferred for all five childhood immunization doses; however, there are few data on the safety and immunogenicity of five consecutive doses. This study compared a fifth dose of an acellular and a whole cell pertussis vaccine in 4- to 6-year-old children previously immunized with four doses of acellular or whole cell pertussis vaccine. STUDY DESIGN: In a double blind, multicenter study, 366 healthy children were randomly allocated to receive a single injection of a 5-component acellular or a whole cell pertussis vaccine, each combined with diphtheria and tetanus toxoids and inactivated poliovirus vaccine. RESULTS: Although injection site redness > or =50 mm and swelling > or =50 mm were common in children who had received five doses of acellular (50% and 48.1%, respectively) or whole cell (66.2% and 59.7%) pertussis vaccine, limb soreness and limitation of motion were less frequently reported after acellular (1.9% and 0%) than after whole cell (49.2% and 36.3%; P < 0.0001) pertussis vaccine. Pre-fifth dose antipertussis antibody titers were higher in children who previously had received four doses of acellular pertussis vaccine. Postimmunization antibody titers against pertussis toxin, filamentous hemagglutinin, pertactin and tetanus toxin were higher in recipients of five doses of acellular pertussis vaccine, whereas antibody titers to diphtheria toxin, pertussis fimbriae and poliovirus serotypes were higher in recipients of five doses of the whole cell pertussis vaccine (P < 0.05 for all comparisons). CONCLUSIONS: A regimen consisting of five doses of a five-component acellular pertussis combination vaccine is safe and immunogenic in pre-school children. Local adverse reactions are common but are less painful and activity-limiting than a regimen of five doses of a whole cell pertussis vaccine.

Extended follow-up of antibody levels and antigen responsiveness after 2 Haemophilus influenzae type b conjugate vaccines.

Although immunization programs with Haemophilus influenzae type b (Hib) conjugate vaccines have dramatically reduced disease incidence, few data are available regarding the duration of protection after vaccination. We measured serum anti-polyribosylribitol phosphate (PRP) levels in healthy 4- to 5- year-old children previously given 4 doses of PRP-T vaccine (at 2, 4, 6, and 18 months) or 1 dose of PRP-D vaccine (at 19 months) during clinical trials to assess antibody persistence. Concurrent with other preschool immunizations, half of the children were randomly assigned to receive a PRP-T booster immunization to assess responsiveness. Among 136 subjects who were primed with PRP-D, the baseline geometric mean concentration of antibody was 0.7 microg/mL (95% CI 0.5 to 0.9). Concentrations were <0.15 microg/mL in 24 (17.6%) subjects. Among 212 children who were primed with PRP-T, the geometric mean concentration was 2.2 microg/mL (95% CI 1.9 to 2.5) (P <.001). Only 2 (0.9%) had concentrations <0.15 microg/mL. Four weeks after PRP-T immunization, geometric mean concentrations had increased to 98.4 and 102.0 microg/mL, respectively. Responses were strong even in those with low or undetectable preimmunization antibody levels. Spontaneous increases in antibody levels were seen in 9 (5.2%) of 172 subjects not given additional PRP-T. We concluded that among 4- to 5-year-olds, anti-PRP levels remained above 0.15 microg/mL in nearly all children after PRP-T priming and in most after PRP-D priming, and that both groups were able to respond vigorously to restimulation, consistent with persistent immune memory.

Acellular pertussis vaccine as a booster dose for seventeen- to nineteen-month-old children immunized with either whole cell or acellular pertussis vaccine at two, four and six months of age.

The safety and immunogenicity of two formulations of an acellular pertussis vaccine as a booster at 17 to 19 months of age were assessed in children immunized at 2, 4 and 6 months of age with acellular or whole cell pertussis vaccine. In Study I 86 children primed with a five-component acellular vaccine combined with diphtheria and tetanus toxoids or with a whole cell pertussis-diphtheria-tetanus vaccine were boosted with the same vaccine. Local reactions (64% vs. 93%; relative risk, 0.7; 95% confidence interval, 0.5 to 0.9) and systemic reactions (68% vs. 97%; relative risk, 0.7; 95% confidence interval, 0.5 to 0.9) were less common after the fourth dose of acellular vaccine than after the fourth dose of whole cell vaccine. In Study II 96 children primed with either an acellular or whole cell pertussis vaccine were boosted with an acellular vaccine. Local adverse reactions after booster immunization with acellular vaccine were more common in children primed with acellular vaccine than those primed with whole cell vaccine (68% vs. 33%; relative risk, 2.1; 95% confidence interval, 1.3 to 3.3). Antibody response to pertussis toxin, filamentous hemagglutinin and fimbriae were higher before and 1 month after the booster dose in children primed with the acellular vaccine. We conclude that the acellular pertussis vaccine is safe and immunogenic when used for the booster dose in children primed with either whole cell or acellular vaccine but is associated with local reactions.

An outbreak of diarrhea due to verotoxin-producing Escherichia coli in the Canadian Northwest Territories.

In the summer of 1991 a large outbreak of Escherichia coli O157:H7 associated diarrhea occurred in 6 Inuit communities in the Canadian Northwest Territories. The total population of these communities is 5,292. Of the 521 individuals who developed diarrhea, 152 (29%) were positive for E. coli O157:H7 on stool culture or positive by verotoxin analysis. Median age was 6 years. The attack rate for children < 1 year was 43% in the major affected community of Arviat. Hemolytic-uremic syndrome (HUS) developed in 22 cases, and 2 patients died. Asymptomatic stool carriage of verotoxin-producing E. coli (VTEC) 2-5 weeks after diarrheal illness was noted in 4/28 persons followed prospectively. Epidemic curves, case-control studies and phage type testing suggested person-to-person transmission. The original source of infection was not identified, though a food source was suspected. VTEC were detected in 6 food samples (minced beef and caribou) taken from retail outlets and homes. Primary prevention of infection through health education and promotion activities, as well as long-term follow-up of HUS survivors, are indicated in this population.