Multicenter Selective Lymphadenectomy Trial-I confirms the central role of sentinel node biopsy in contemporary melanoma management: response to 'No survival benefit for patients with melanoma undergoing sentinel lymph node biopsy: critical appraisal of the Multicenter Selective Lymphadenectomy Trial-I final report'.

Sentinel lymph node (SLN) biopsy has become a standard procedure for many patients with melanoma and is recommended in numerous national and professional melanoma guidelines. The Multicenter Selective Lymphadenectomy Trial (MSLT-1) confirms earlier large database studies and prospective clinical trials in demonstrating the independent and unequalled prognostic value of the SLN. It also demonstrates the ability of biopsy-directed management to provide effective regional disease control with the least possible morbidity. These benefits are not in question and provide ample justification for the procedure, even without evidence of a survival benefit. However, MSLT-1 also provides strong evidence of a substantial reduction in the risk of melanoma death for patients with intermediate thickness melanomas who harbour occult nodal metastases at the time of presentation. Denying appropriately selected patients with melanoma the opportunity to undergo SLN biopsy is no longer reasonable or acceptable.

Generation and characterization of phage-GnRH chemical conjugates for potential use in cat and dog immunocontraception.

Overpopulation of cats and dogs is a serious worldwide problem that demands novel, safe and cost-effective solutions. The objective of this study was to generate and characterize phage-peptide conjugates with gonadotropin-releasing hormone (GnRH) for potential use as an immunocontraceptive. A filamentous phage vector f5-8 with wild-type phage coat proteins was used as a carrier for construction of chemical conjugates with GnRH, a peptide that acts as a master reproductive hormone. In such conjugates, the phage body plays the role of a carrier protein, while multiple copies of GnRH peptide stimulate production of neutralizing anti-GnRH antibodies potentially leading to contraceptive effects. To generate the constructs, four different GnRH-based peptides were synthesized and conjugated to phage particles in a two-step procedure: (i) peptides were reacted with phage to form a conjugate using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride chemistry (EDC) and (ii) the conjugates were separated from remaining free peptides by dialysis. Formation and specificity of phage-GnRH conjugates were confirmed by three independent methods: spectrophotometry, electron microscopy and ELISA. When the conjugates were tested for interaction with sera collected from cats and dogs immunized with GnRH-based vaccines in independent studies, strong specific ELISA signals were obtained, suggesting the potential use of the conjugates for cat and dog immunosterilization. The ability of the conjugates to stimulate production of anti-GnRH antibodies in vivo was evaluated in mice. While optimization of dose, immunization route and adjuvant still requires investigation, our preliminary results demonstrated the presence of anti-GnRH antibodies in sera of mice immunized with such conjugates. Fertility trials in cats and dogs will be needed to evaluate contraceptive potentials of the phage-GnRH peptide chemical conjugates.

Antibody-catalyzed porphyrin metallation.

An antibody elicited to a distorted N-methyl porphyrin catalyzed metal ion chelation by the planar porphyrin. At fixed Zn2+ and Cu2+ concentrations, the antibody-catalyzed reaction showed saturation kinetics with respect to the substrate mesoporphyrin IX (2) and was inhibited by the hapten, N-methylmesoporphyrin IX (1). The turnover number of 80 hour-1 for antibody-catalyzed metallation of 2 with Zn2+ compares with an estimated value of 800 hour-1 for ferrochelatase. The antibody also catalyzed the insertion of Co2+ and Mn2+ into 2, but it did not catalyze the metallation of protoporphyrin IX (3) or deuteroporphyrin IX (4). The antibody has high affinity for several metalloporphyrins, suggesting an approach to developing antibody-heme catalysts for redox or electron transfer reactions.

Imitation of Escherichia coli aspartate receptor signaling in engineered dimers of the cytoplasmic domain.

Transmembrane signaling by bacterial chemotaxis receptors appears to require a conformational change within a receptor dimer. Dimers were engineered of the cytoplasmic domain of the Escherichia coli aspartate receptor that stimulated the kinase CheA in vitro. The folding free energy of the leucine-zipper dimerization domain was harnessed to twist the dimer interface of the receptor, which markedly affected the extent of CheA activation. Response to this twist was attenuated by modification of receptor regulatory sites, in the same manner as adaptation resets sensitivity to ligand in vivo. These results suggest that the normal allosteric activation of the chemotaxis receptor has been mimicked in a system that lacks both ligand-binding and transmembrane domains. The most stimulatory receptor dimer formed a species of tetrameric size.

Spectrophotometry of pluto-charon mutual events: individual spectra of pluto and charon.

Time-resolved spectra of the 3 March and 4 April 1987 mutual events of Pluto and its satellite Charon were obtained with spectral coverage from 5,500 to 10,000 angstroms with 25 angstrom spectral resolution. Since both events were total occultations of Charon by Pluto, spectra were obtained of the anti-Charon-facing hemisphere of Pluto, with no contribution from Charon during totality. On 4 April, a combined spectrum of Pluto and Charon immediately before first contact was also obtained. The spectrum of the Pluto-facing hemisphere of Charon was extracted by differencing the pre-event and totality spectra. The spectra were reduced to reflectances by ratioing them to spectra of solar analog stars. Charon has a featureless reflectance spectrum, with no evidence of methane absorption. Charon's reflectance appears neutral in color and corresponds to a geometric albedo of approximately 0.37 at 6000 angstroms. The Pluto reflectance spectrum displays methane absorption bands at 7300, 7900, 8400, 8600, and 8900 angstroms and is red in color, with a geometric albedo of approximately 0.56 at 6000 angstroms. The signal-to-noise ratios of the eclipse spectra were not high enough to unambiguously identify the weaker methane band at 6200 angstroms.

Nerve transfers for peripheral nerve injury in the upper limb: a case-based review.

Nerve transfer has become a common and often effective reconstructive strategy for proximal and complex peripheral nerve injuries of the upper limb. This case-based discussion explores the principles and potential benefits of nerve transfer surgery and offers in-depth discussion of several established and valuable techniques including: motor transfer for elbow flexion after musculocutaneous nerve injury, deltoid reanimation for axillary nerve palsy, intrinsic re-innervation following proximal ulnar nerve repair, and critical sensory recovery despite non-reconstructable median nerve lesions.

Lamellar pro-inflammatory cytokine expression patterns in laminitis at the developmental stage and at the onset of lameness: innate vs. adaptive immune response.

REASONS FOR PERFORMING STUDY: Recent research has indicated that inflammation plays a role in the early stages of laminitis and that, similar to organ failure in human sepsis, early inflammatory mechanisms may lead to downstream events resulting in lamellar failure. Characterisation of the type of immune response (i.e. innate vs. adaptive) is essential in order to develop therapeutic strategies to counteract these deleterious events. OBJECTIVES: To quantitate gene expression of pro-inflammatory cytokines known to be important in the innate and adaptive immune response during the early stages of laminitis, using both the black walnut extract (BWE) and oligofructose (OF) models of laminitis. METHODS: Real-time qPCR was used to assess lamellar mRNA expression of interleukins-1beta, 2, 4, 6, 8, 10, 12 and 18, and tumour necrosis factor alpha and interferon gamma at the developmental stage and at the onset of lameness. RESULTS: Significantly increased lamellar mRNA expression of cytokines important in the innate immune response were present at the developmental stage of the BWE model, and at the onset of acute lameness in both the BWE model and OF model. Of the cytokines characteristic of the Th1 and Th2 arms of the adaptive immune response, a mixed response was noted at the onset of acute lameness in the BWE model, whereas the response was skewed towards a Th1 response at the onset of lameness in the OF model. CONCLUSIONS: Lamellar inflammation is characterised by strong innate immune response in the developmental stages of laminitis; and a mixture of innate and adaptive immune responses at the onset of lameness. POTENTIAL RELEVANCE: These results indicate that anti-inflammatory treatment of early stage laminitis (and the horse at risk of laminitis) should include not only therapeutic drugs that address prostanoid activity, but should also address the marked increases in lamellar cytokine expression.

The 67P/Churyumov-Gerasimenko observation campaign in support of the Rosetta mission.

We present a summary of the campaign of remote observations that supported the European Space Agency's Rosetta mission. Telescopes across the globe (and in space) followed comet 67P/Churyumov-Gerasimenko from before Rosetta's arrival until nearly the end of the mission in September 2016. These provided essential data for mission planning, large-scale context information for the coma and tails beyond the spacecraft and a way to directly compare 67P with other comets. The observations revealed 67P to be a relatively 'well-behaved' comet, typical of Jupiter family comets and with activity patterns that repeat from orbit to orbit. Comparison between this large collection of telescopic observations and the in situ results from Rosetta will allow us to better understand comet coma chemistry and structure. This work is just beginning as the mission ends-in this paper, we present a summary of the ground-based observations and early results, and point to many questions that will be addressed in future studies.This article is part of the themed issue 'Cometary science after Rosetta'.

Zoned immune suppression of lymph nodes draining malignant melanoma: histologic and immunohistologic studies.

Node-to-node heterogeneity of reaction and recognizable patterns of reaction in node groups draining melanoma were sought. Nodes from 72 patients undergoing lymphadenectomy for high-risk, primary melanoma (57) or node-spread melanoma (15) were accurately oriented to the nearest melanoma. Reactivity of paracortex, follicular areas, and sinuses was assessed on a 0-3+ scale. Reactivity was prominent in paracortex and sinuses but varied from node to node within node groups. Nodes nearest to tumor showed least reaction; nodes at intermediate distances from tumor were most reactive, while those farthest away showed mostly little reaction. Variation of nodal reaction that correlated with the node position relative to the nearest melanoma (zoned reaction) was seen in 92% of patients with nodal metastases of melanoma and in 64% of patients with primary malignant melanoma. Follicular and sinusoidal reactions showed no significant zoning. S-100 protein-positive paracortical dendritic cells (PDCs) in tumor-oriented nodes were quantified. PDCs were infrequent in nodes partly replaced by melanoma or located near to melanoma but were numerous in nodes located farther from tumor. Changes of nodal activity (relative stimulation or suppression) correlate with the distance of the node from the nearest deposit of primary or metastatic melanoma.

Frequency and distribution of occult micrometastases in lymph nodes of patients with non-small-cell lung carcinoma.

BACKGROUND: Accurate assessment of the presence and absence of tumor in the regional lymph nodes is critical in assessment of prognosis for patients with lung cancer. Development of sensitive immunohistochemical techniques and specific monoclonal antibodies has increased our capacity, in melanoma and breast cancer, to detect small groups of tumor cells or even single tumor cells in lymph nodes that appear to be tumor free in conventionally stained sections. PURPOSE: This retrospective study was designed to assess whether use of a polyclonal antikeratin reagent in immunohistochemical analysis offers any advantage over conventional histopathology in detection of regional lymph node metastases in non-small-cell lung cancer. METHODS: Paraffin-embedded tissue sections from regional lymph nodes of 65 patients with non-small-cell lung cancer were studied. We examined tissue from 588 nodes of 60 patients with a diagnosis of disease confined to the lung and from 72 nodes of five patients with a diagnosis of metastasis to some nodes. A polyclonal antikeratin antibody was applied to the lymph node tissue sections, using the avidin-biotin complex immunoperoxidase technique. RESULTS: Single tumor cells and small clusters of tumor cells (occult micrometastases) not visible on routine evaluation were readily detected in 38 (63%) of the 60 patients whose nodes appeared to be negative on examination of hematoxylin-eosin-stained slides. In the five patients with a diagnosis of node-positive non-small-cell lung cancer, five (10%) of 51 nodes that were tumor free on conventional examination contained metastases. Metastatic tumor cells were most often located in the subcapsular or medullary sinuses. The lymph nodes that contained occult tumor cells were those located nearest to the tumor, mainly in peribronchial and hilar locations. The median survival of patients with occult metastases (1977 days) was shorter than that of patients whose nodes contained no tumor (2456 days) but was longer than that of patients whose nodes contained metastases detectable on hematoxylin-eosin-stained slides (927 days). CONCLUSIONS: Our results suggest that, in patients with non-small-cell lung cancer, metastatic involvement of regional lymph nodes is more frequent than was previously determined by the conventional histologic method and substantially more frequent than in other tumor types, such as melanoma and breast cancer. IMPLICATIONS: The high frequency of occult nodal metastases in non-small-cell lung cancer makes it clear that, without immunohistochemistry, disease is understaged in many patients. Therefore, it seems essential that immunohistochemical evaluation of the lymph nodes be undertaken in clinical trials.

Variations in functional immunocompetence of individual tumor-draining lymph nodes in humans.

In studying the histology and immunohistology of tumor-draining lymph nodes, we observed that nodes close to tumor showed reduced paracortical activity relative to those further away. To assess whether this reduction was paralleled by alterations in the functional activity of nodes, we examined the immunocompetence of individually oriented tumor-draining lymph nodes. We assessed the unstimulated activity of lymph node lymphocytes and their responses to phytohemagglutinin, interleukin 2, and one-way alloantigenic stimulation (mixed lymphocyte reaction) in vitro. Regional nodes from patients with malignant melanoma or breast cancer were classified as proximal, intermediate, or distal relative to tumor. Node groups with and without tumor metastases in them were studied. Significant variations in [3H]thymidine uptake were noted with the unstimulated and stimulated lymphocytes of different lymph nodes from the same individual. Nodes near to tumor were less responsive than those located further away; some of the latter showed relative immunostimulation. Tumor-draining node groups thus demonstrated a nonrandom variation in the strength of reaction of individual nodes. There are zones of low and high lymph node reactivity, related to the position of each node relative to tumor. Tumor-derived immunosuppressive products and/or immunoregulation down-regulates lymph node functional activity, creating conditions that may permit the survival of tumor cells and the establishment of metastases. It is suggested that immunosuppression of nodes nearest to tumor may facilitate the early establishment of metastases.

Suppressor cell activity in melanoma-draining lymph nodes.

We previously demonstrated that the cells of lymph nodes near to a melanoma respond well to stimulation by mitogens, alloantigens, and interleukin 2 than do nodes further away. In this study we examined suppressor T-cell activity in nodes at different distances from primary melanoma, using a concanavalin A (Con A) suppressor cell assay. Tumor-free regional nodes were classified as proximal, intermediate, and distal relative to primary melanoma. Lymph node lymphocytes (LNL) were stimulated with 50 micrograms/ml Con A for 48-72 h, inactivated, and then mixed with autologous peripheral blood lymphocytes. The peripheral blood lymphocyte-LNL mixtures were stimulated with phytohemagglutinin for 3 days. Proliferation was measured by [3H]thymidine uptake during the final 18 h of culture. In 13 patients, Con A-treated LNL from nodes near to tumor were more suppressive of the peripheral blood lymphocyte response to phytohemagglutinin than those from nodes located further from tumor. T-lymphocyte subset assessment before and after Con A treatment of LNL showed no significant changes in T4:T8 ratios. Con A-induced suppressor cells could be maintained in culture in the presence of recombinant interleukin 2 and retained their suppressive activity. LNL not exposed to Con A and maintained in culture with interleukin 2 did not show suppressor cell activity. Suppressor cell activity thus contributes to the weak immune reactivity of lymph nodes near to melanoma.

Lymphocyte subset alterations in nodes regional to human melanoma.

The lymphocyte subpopulations in tumor-draining lymph nodes of melanoma patients were determined using two-color flow cytometry. Data were analyzed according to: (a) the staging of the melanoma; (b) whether or not the nodes contained tumor; and (c) their distance from the primary tumor. Compared with Stage I patients (without metastasis), uninvolved nodes of stage II patients (with nodal metastases) had a significant decrease in helper/inducer (CD4+) T-cells (P less than 0.001), with a corresponding increase in cytotoxic/suppressor (CD8+) cells (P less than 0.001) and Leu 19+ natural killer (CD56+) cells (P less than 0.01). In some patients the presence of tumor within a node was associated with a large decrease in CD3+ total T-cells, whereas in others tumor involvement had little influence on lymphocyte phenotypes. When analyzed by distance from the primary tumor, nodes closest to tumor in Stage I patients contained a smaller percentage of CD19+ B-cells. In Stage II, tumor-free nodes nearest to tumor showed an increase in CD19+ cells, but statistical significance was not reached. CD56+ natural killer cells increased progressively in nodes near tumor and were more numerous in Stage II uninvolved nodes compared with Stage I nodes. Alterations in phenotypically defined lymph node lymphocytes occur in nodes regional to melanoma as the disease progresses, as growth of metastases occurs, and in tumor-free nodes nearest to tumor. These alterations may be essential to the establishment and progression of metastases.

Data-driven classification of bipolar I disorder from longitudinal course of mood.

The Diagnostic and Statistical Manual of Mental Disorder (DSM) classification of bipolar disorder defines categories to reflect common understanding of mood symptoms rather than scientific evidence. This work aimed to determine whether bipolar I can be objectively classified from longitudinal mood data and whether resulting classes have clinical associations. Bayesian nonparametric hierarchical models with latent classes and patient-specific models of mood are fit to data from Longitudinal Interval Follow-up Evaluations (LIFE) of bipolar I patients (N=209). Classes are tested for clinical associations. No classes are justified using the time course of DSM-IV mood states. Three classes are justified using the course of subsyndromal mood symptoms. Classes differed in attempted suicides (P=0.017), disability status (P=0.012) and chronicity of affective symptoms (P=0.009). Thus, bipolar I disorder can be objectively classified from mood course, and individuals in the resulting classes share clinical features. Data-driven classification from mood course could be used to enrich sample populations for pharmacological and etiological studies.

Influence of extended aging on beef quality characteristics and sensory perception of steaks from the biceps femoris and semimembranosus.

The objective was to determine the influence of post-fabrication aging (2, 14, 21, 42, and 63days) on beef quality characteristics and consumer sensory perception of biceps femoris (BF) and semimembranosus (SM) steaks. Lipid oxidation and aerobic plate counts increased (P<0.05) with longer aging periods and retail display times. An aging period by day of retail display interaction (P<0.05) was observed for a* and b* values of the BF and SM. Warner-Bratzler shear force values decreased (P<0.05) with longer aging for the SM, while no difference was observed for the BF. Consumer panel results revealed that longer aging periods increased (P<0.05) acceptability of the SM, tenderness of both muscles, and tended to increase (P=0.07) juiciness of the SM. Our results show that extended aging reduces retail color stability yet has positive effects on consumer perception of tenderness of both muscles and overall acceptability of the SM.

Intraoperative lymphatic mapping and selective cervical lymphadenectomy for early-stage melanomas of the head and neck.

PURPOSE: We developed intraoperative lymphatic mapping with selective lymphadenectomy (SLND) to identify micrometastatic spread of cutaneous melanoma to regional lymph nodes. This study was undertaken to assess the sensitivity and specificity of our technique in patients with clinical stage I (CS-I) melanoma of the head or neck. PATIENTS AND METHODS: Seventy-two CS-I melanoma patients underwent intraoperative lymphatic mapping of primary cutaneous melanomas located on the head, neck, or upper chest/back draining to the neck. Key (sentinel) cervical lymph nodes in the regional lymphatic drainage basin were identified, selectively excised during SLND, and examined for microscopic evidence of tumor cells. If these sentinel nodes were tumor-negative, the surgery was concluded; if the sentinel nodes were tumor-positive, all nodes in the drainage basin were removed during en bloc lymphadenectomy (LND). RESULTS: Intraoperative lymphatic mapping identified sentinel nodes in 90% of the regional drainage basins. Fifteen percent of these nodes were tumor-positive, indicating the need for LND. There were no false-negative sentinel nodes, and extended follow-up showed no local nodal recurrences in patients whose sentinel-node histology did not indicate the need for LND. CONCLUSION: Intraoperative lymphatic mapping and SLND is a minimally invasive and highly accurate screening technique for determining which patients with CS-I head and neck melanomas have subclinical node metastases and therefore might benefit from cervical LND.

Dobutamine thallium-201 tomography for evaluating patients with suspected coronary artery disease unable to undergo exercise or vasodilator pharmacologic stress testing.

OBJECTIVES: The aim of this study was to assess the feasibility, safety and diagnostic accuracy of a high dose dobutamine infusion in conjunction with thallium-201 single-photon emission computed tomography in 144 patients (72 men and 72 women with a mean age of 65 +/- 10 years) unable to perform exercise or pharmacologic vasodilator stress testing. BACKGROUND: Dobutamine increases myocardial oxygen consumption by increasing heart rate, contractility and arterial blood pressure. In addition, it causes myocardial blood flow heterogeneity and thus may be a useful stress for noninvasive detection of coronary artery disease. METHODS: Dobutamine was administered intravenously at incremental doses of 5, 10, 20, 30 and up to 40 micrograms/kg per min at 3-min intervals. After 1 min of the maximal dose, 3 mCi of thallium-201 was injected and the infusion was continued for an additional 2 min. Thallium-201 tomography was performed 5 to 10 min after termination of the infusion and 4 h later. The images were visually assessed for the presence and vascular location of perfusion defects and the extent of thallium redistribution. Coronary angiography was performed in 84 patients, with a > 50% stenosis considered significant. RESULTS: Dobutamine significantly (p = 0.0001) increased the heart rate (from 75 +/- 14 beats/min to 120 +/- 23 beats/min), systolic blood pressure (from 136 +/- 23 mm Hg to 148 +/- 35 mm Hg) and the rate-pressure product (from 10,144 +/- 2,517 to 17,858 +/- 4,349) from baseline to peak infusion rate, respectively. Most patients (75%) experienced side effects during the infusion, but 74% tolerated a dobutamine dose of 40 micrograms/kg per min and 97% a dose of 30 micrograms/kg per min. The more common side effects were typical (26%) and atypical (5%) chest pain, palpitation (29%), flushing (14%), headache (14%) and dyspnea (14%). The overall sensitivity of dobutamine tomography was 86% in the patients who underwent coronary angiography and 84% in those with single-vessel, 82% in those with double-vessel and 100% in those with triple-vessel disease. Seventy-eight percent of vessels with severe (> or = 70%) stenoses were identified with dobutamine tomography. The specificity of dobutamine tomography was 90% for patients and 86% for individual vessels. CONCLUSIONS: A high dose dobutamine infusion in conjunction with thallium tomography appears to be a well tolerated and accurate method for diagnosing coronary artery disease in patients unable to perform exercise or vasodilator pharmacologic stress testing.

Protein-protein interfaces: mimics and inhibitors.

Many biological processes are mediated by specific molecular recognition between proteins. However, the thermodynamic and structural 'rules' for such recognition are incompletely understood, as is the potential for inhibition by small molecules. Recent progress has included the discovery of small-molecule inhibitors for several targets important in cancer.

Antagonists of protein-protein interactions.

Protein-protein interactions are often attractive, but not straightforward, targets for disease therapy. Two strategies for identifying inhibitors of these interactions, peptide phage display and high-throughput screening, have recently shown new promise.

Influence of extended aging on beef quality characteristics and sensory perception of steaks from the gluteus medius and longissimus lumborum.

The objective was to determine the influence of post-fabrication aging (2, 14, 21, 42, and 63 days) on beef quality characteristics and consumer sensory perception of gluteus medius (GM) and longissimus lumborum (LL) steaks. Lipid oxidation and aerobic plate counts increased (P<0.05) with longer aging periods and retail display times. An aging period by day of retail display interaction (P<0.05) was observed for a* and b* values for both muscles and L* values for the LL. Warner-Bratzler shear force values decreased (P<0.05) with longer aging for the LL, while no difference was observed for the GM. Consumer panel results demonstrated that longer aging periods increased (P<0.05) tenderness of both muscles. Our results indicate that extended aging reduces retail color stability yet has positive effects on consumer perception of tenderness of beef loin muscles.