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Trichorhinophalangeal syndrome 1 (TRPS1) is a nuclear protein highly expressed in breast epithelial cells. TRPS1 immunohistochemistry (IHC) has been suggested as a breast cancer marker. To determine the diagnostic and prognostic utility of TRPS1 IHC, tissue microarrays containing 19,201 samples from 152 different tumor types and subtypes were analyzed. GATA3 IHC was performed in a previous study. TRPS1 staining was seen in 86 of 152 tumor categories with 36 containing at least one strongly positive case. TRPS1 staining predominated in various types of breast carcinomas (51%-100%), soft tissue tumors (up to 100%), salivary gland tumors (up to 46%), squamous cell carcinomas (up to 35%), and gynecological cancers (up to 40%). TRPS1 positivity occurred in 1.8% of 1083 urothelial neoplasms. In invasive breast carcinoma of no special type, low TRPS1 expression was linked to high grade (P= 0.0547), high pT (P< 0.0001), nodal metastasis (P= 0.0571), loss of estrogen receptor and progesterone receptor expression (P< 0.0001 each), and triple-negative status (P< 0.0001) but was unrelated to patient survival (P= 0.8016). In squamous cell carcinomas from 11 different sites, low TRPS1 expression was unrelated to tumor phenotype. Positivity for both TRPS1 and GATA3 occurred in 47.4% to 100% of breast cancers, up to 30% of salivary gland tumors, and 29 (0.3%) of 9835 tumors from 134 other cancer entities. TRPS1 IHC has high utility for the identification of cancers of breast (or salivary gland) origin, especially in combination with GATA3. The virtual absence of TRPS1 positivity in urothelial neoplasms is useful for the distinction of GATA3-positive urothelial carcinoma from breast cancer.

In maritime traffic surveillance, detecting illegal activities, such as illegal fishing or transshipment of illicit products is a crucial task of the coastal administration. In the open sea, one has to rely on Automatic Identification System (AIS) message transmitted by on-board transponders, which are captured by surveillance satellites. However, insincere vessels often intentionally shut down their AIS transponders to hide illegal activities. In the open sea, it is very challenging to differentiate intentional AIS shutdowns from missing reception due to protocol limitations, bad weather conditions or restricting satellite positions. This paper presents a novel approach for the detection of abnormal AIS missing reception based on self-supervised deep learning techniques and transformer models. Using historical data, the trained model predicts if a message should be received in the upcoming minute or not. Afterwards, the model reports on detected anomalies by comparing the prediction with what actually happens. Our method can process AIS messages in real-time, in particular, more than 500 Millions AIS messages per month, corresponding to the trajectories of more than 60 000 ships. The method is evaluated on 1-year of real-world data coming from four Norwegian surveillance satellites. Using related research results, we validated our method by rediscovering already detected intentional AIS shutdowns.

Background: Depending on the therapy approach and disease background, the heterogeneity of muscular tissues complicates the development of targeted gene therapy, where either expression in all muscle types or restriction to only one muscle type is warranted. Muscle specificity can be achieved using promotors mediating tissue specific and sustained physiological expression in the desired muscle types but limited activity in non-targeted tissue. Several muscle specific promotors have been described, but direct comparisons between them are lacking.Objective: Here we present a direct comparison of muscle specific Desmin-, MHCK7, microRNA206- and Calpain3 promotor.Methods: To directly compare these muscle specific promotors we utilized transfection of reporter plasmids using an in vitromodel based on electrical pulse stimulation (EPS) to provoke sarcomere formation in 2D cell culture for quantification of promotor activities in far differentiated mouse and human myotubes.Results: We found that Desmin- and MHCK7 promotors showed stronger reporter gene expression levels in proliferating and differentiated myogenic cell lines than miR206 and CAPN3 promotor. However, Desmin and MHCK7 promotor promoted gene expression also cardiac cells whereas miR206 and CAPN3 promotor expression was restricted to skeletal muscle.Conclusions: Our results provides direct comparison of muscle specific promotors with regard to expression strengths and specificity as this is important feature to avoid undesired transgene expression in non-target muscle cells for a desired therapy approach.

Depending on the therapy approach and disease background, the heterogeneity of muscular tissues complicates the development of targeted gene therapy, where either expression in all muscle types or restriction to only one muscle type is warranted. Muscle specificity can be achieved using promotors mediating tissue specific and sustained physiological expression in the desired muscle types but limited activity in non-targeted tissue. Several muscle specific promotors have been described, but direct comparisons between them are lacking. Here we present a direct comparison of muscle specific Desmin-, MHCK7, microRNA206- and Calpain3 promotor. To directly compare these muscle specific promotors we utilized transfection of reporter plasmids using an in vitromodel based on electrical pulse stimulation (EPS) to provoke sarcomere formation in 2D cell culture for quantification of promotor activities in far differentiated mouse and human myotubes. We found that Desmin- and MHCK7 promotors showed stronger reporter gene expression levels in proliferating and differentiated myogenic cell lines than miR206 and CAPN3 promotor. However, Desmin and MHCK7 promotor promoted gene expression also cardiac cells whereas miR206 and CAPN3 promotor expression was restricted to skeletal muscle. Our results provides direct comparison of muscle specific promotors with regard to expression strengths and specificity as this is important feature to avoid undesired transgene expression in non-target muscle cells for a desired therapy approach.

To study the extent of heterogeneity of mesothelin overexpression in primary ovarian cancers and their peritoneal and lymph node metastases, a tissue microarray (TMA) was constructed from multiple sites of 220 ovarian cancers and analyzed by immunohistochemistry. One tissue core each was taken from up to 18 different tumor blocks per cancer, resulting in a total of 2460 tissue spots from 423 tumor sites (188 primary cancers, 162 peritoneal carcinosis, and 73 lymph node metastases). Positive mesothelin expression was found in 2041 of the 2342 (87%) arrayed tissue spots and in 372 of the 392 (95%) tumor sites that were interpretable for mesothelin immunohistochemistry. Intratumoral heterogeneity was found in 23% of 168 primary cancer sites interpretable for mesothelin and decreased to 12% in 154 peritoneal carcinosis and to 6% in 71 lymph node metastases (P<0.0001). Heterogeneity between the primary tumor and matched peritoneal carcinosis was found in 16% of 102 cancers with interpretable mesothelin results. In these cancers, the mesothelin status switched from positive in the primary tumor to negative in the peritoneal carcinosis (3 cancers) in or vice versa (2 cancers), or a mixture of positive and negative peritoneal carcinoses was found (11 cancers). No such switch was seen between the mesothelin-interpretable primary tumors and their nodal metastases of 59 cancers, and only 1 mesothelin-positive tumor had a mixture of positive and negative lymph node metastases. In conclusion, mesothelin expression is frequent and highly homogeneous in ovarian cancer.

CTLA-4 is an inhibitory immune checkpoint receptor and a negative regulator of anti-tumor T-cell function. This study is aimed for a comparative analysis of CTLA-4+cells between different tumor entities. To quantify CTLA-4+cells, 4582 tumor samples from 90 different tumor entities as well as 608 samples of 76 different normal tissue types were analyzed by immunohistochemistry in a tissue microarray format. Two different antibody clones (MSVA-152R and CAL49) were validated and quantified using a deep learning framework for automated exclusion of unspecific immunostaining. Comparing both CTLA-4 antibodies revealed a clone dependent unspecific staining pattern in adrenal cortical adenoma (63%) for MSVA-152R and in pheochromocytoma (67%) as well as hepatocellular carcinoma (36%) for CAL49. After automated exclusion of non-specific staining reaction (3.6%), a strong correlation was observed for the densities of CTLA-4+lymphocytes obtained by both antibodies (r= 0.87; p< 0.0001). A high CTLA-4+cell density was linked to low pT category (p< 0.0001), absent lymph node metastases (p= 0.0354), and PD-L1 expression in tumor cells or inflammatory cells (p< 0.0001 each). A high CTLA-4/CD3-ratio was linked to absent lymph node metastases (p= 0.0295) and to PD-L1 positivity on immune cells (p= 0.0026). Marked differences exist in the number of CTLA-4+lymphocytes between tumors. Analyzing two independent antibodies by a deep learning framework can facilitate automated quantification of immunohistochemically analyzed target proteins such as CTLA-4.

Carboxypeptidase A1 (CPA1) is a zinc metalloprotease that is produced in pancreatic acinar cells and plays a role in cleaving C-terminal branched-chain and aromatic amino acids from dietary proteins. This study assessed the utility of immunohistochemical CPA1 staining for diagnosing pancreatic acinar cell carcinoma (ACC). A total of 12,274 tumor samples from 132 different tumor types and subtypes as well as 8 samples each of 76 different normal tissue types were interpretable by immunohistochemistry in a tissue microarray format. CPA1 was strongly expressed in acinar cells of all normal pancreas samples but not in any other normal tissues. CPA1 immunostaining was detected in 100% of 11 pancreatic ACCs and 1 mixed acinar endocrine carcinoma, but absent in 449 pancreatic ductal adenocarcinomas, 75 adenocarcinomas of the ampulla Vateri, and 11,739 other evaluable cancers from 128 different tumor entities. A weak to moderate diffuse staining of epithelial and stromal cells of cancer tissues immediately adjacent to non-neoplastic pancreatic acinar cells often occurred and was considered to be caused by the diffusion of the highly abundant CPA1 from normal acinar cells that may have suffered some autolytic cell damage. In conclusion, our data show that CPA1 is a highly sensitive and largely specific marker for normal and neoplastic pancreatic acinar cells. CPA1 immunohistochemistry greatly facilitates the otherwise often difficult diagnosis of pancreatic ACC.

CTLA-4 is an inhibitory immune checkpoint receptor and a negative regulator of anti-tumor T-cell function. This study is aimed for a comparative analysis of CTLA-4+cells between different tumor entities. To quantify CTLA-4+cells, 4582 tumor samples from 90 different tumor entities as well as 608 samples of 76 different normal tissue types were analyzed by immunohistochemistry in a tissue microarray format. Two different antibody clones (MSVA-152R and CAL49) were validated and quantified using a deep learning framework for automated exclusion of unspecific immunostaining. Comparing both CTLA-4 antibodies revealed a clone dependent unspecific staining pattern in adrenal cortical adenoma (63%) for MSVA-152R and in pheochromocytoma (67%) as well as hepatocellular carcinoma (36%) for CAL49. After automated exclusion of non-specific staining reaction (3.6%), a strong correlation was observed for the densities of CTLA-4+lymphocytes obtained by both antibodies (r= 0.87; p< 0.0001). A high CTLA-4+cell density was linked to low pT category (p< 0.0001), absent lymph node metastases (p= 0.0354), and PD-L1 expression in tumor cells or inflammatory cells (p< 0.0001 each). A high CTLA-4/CD3-ratio was linked to absent lymph node metastases (p= 0.0295) and to PD-L1 positivity on immune cells (p= 0.0026). Marked differences exist in the number of CTLA-4+lymphocytes between tumors. Analyzing two independent antibodies by a deep learning framework can facilitate automated quantification of immunohistochemically analyzed target proteins such as CTLA-4.

Junctional epidermolysis bullosa (JEB) is a severe and often lethal genetic disease caused by mutations in genes encoding the basement membrane component laminin-332. Surviving patients with JEB develop chronic wounds to the skin and mucosa, which impair their quality of life and lead to skin cancer. Here we show that autologous transgenic keratinocyte cultures regenerated an entire, fully functional epidermis on a seven-year-old child suffering from a devastating, life-threatening form of JEB. The proviral integration pattern was maintained in vivo and epidermal renewal did not cause any clonal selection. Clonal tracing showed that the human epidermis is sustained not by equipotent progenitors, but by a limited number of long-lived stem cells, detected as holoclones, that can extensively self-renew in vitro and in vivo and produce progenitors that replenish terminally differentiated keratinocytes. This study provides a blueprint that can be applied to other stem cell-mediated combined ex vivo cell and gene therapies.

Focal T lymphocyte aggregates commonly occur in colorectal cancer; however, their biological significance is unknown. To study focal aggregates of T lymphocytes, a deep learning–based framework for automated identification of T cell accumulations (T cell nests) was developed using CD8, PD-1, CD112R, and Ki67 multiplex fluorescence immunohistochemistry. To evaluate the clinical significance of these parameters, a cohort of 523 colorectal cancers with clinical follow-up data was analyzed. Spatial analysis of locally enriched CD8+T cell density and cell-to-cell contacts identified T cell nests in the tumor microenvironment of colorectal cancer. CD112R and PD-1 expressions on CD8+T cells located in T cell nests were found to be elevated compared with those on CD8+T cells in all other tumor compartments (P< .001 each). Although the highest mean CD112R expression on CD8+T cells was observed at the invasive margin, the PD-1 expression on CD8+T cells was elevated in the center of the tumor (P< .001 each). Across all tissue compartments, proliferating CD8+T cells showed higher relative CD112R and PD-1 expressions than those shown by non-proliferating CD8+T cells (P< .001 each). Integration of all available spatial and immune checkpoint expression parameters revealed a superior predictive performance for overall survival (area under the curve, 0.65; 95% CI, 0.60-0.70) compared with the commonly used CD8+tumor-infiltrating lymphocyte density (area under the curve, 0.57; 95% CI, 0.53-0.61; P< .001). Cytotoxic T cells with elevated CD112R and PD-1 expression levels are orchestrated in T cell nests of colorectal cancer and predict favorable patient outcomes, and the spatial nonredundancy underlies fundamental differences between both inhibitory immune checkpoints that provide a rationale for dual anti-CD112R/PD-1 immune checkpoint therapy.

Sporadic inclusion body myositis (sIBM) and polymyositis (PM) are characterized by muscle inflammation, with sIBM showing additional degenerative alterations. In this study we investigated human beta defensins and associated TLRs to elucidate the role of the innate immune system in idiopathic inflammatory myopathies (IIM), and its association with inflammatory and degenerative alterations. Expression levels of human beta-defensin (HBD)-1, HBD-2, HBD-3 and TLR2, 3, 4, 7 and 9 were analysed by quantitative real-time PCR in skeletal muscle tissue. Localization of HBD-3, collagen 6, dystrophin, CD8-positive T-cells, CD-68-positive macrophages, β-amyloid, the autophagy marker LC3, and TLR3 were detected by immunofluorescence and co-localization was quantified. HBD-3 and all TLRs except for TLR9 were overexpressed in both IIM with significant overexpression of TLR3 in sIBM. HBD-3 showed characteristic intracellular accumulations near deposits of β-amyloid, LC3 and TLR3 in sIBM, and was detected in inflammatory infiltrations and macrophages invading necrotic muscle fibres in both IIM. The characteristic intracellular localization of HBD-3 near markers of degeneration and autophagy, and overexpression of endosomal TLR3 in sIBM hint at different pathogenetic mechanisms in sIBM compared with PM. This descriptive study serves as a first approach to the role of the innate immune system in sIBM and PM.

Wound infections caused by multidrug-resistant bacteria are a major issue in wound care. An occlusive dressing delivering an antimicrobial agent to the wound may be advantageous. The objective of this study was to evaluate an occlusive silk membrane loaded with colistin to establish an effective antimicrobial wound dressing against Gram-negative bacteria in vitro and in vivo.

A primary strategy for preventing and treating wound infection in chronic wounds is the use of topical antiseptics and wound irrigating agents. However, their interaction with commonly used wound dressings has not yet been investigated. In this study, the authors analyzed the antimicrobial activity of antiseptics and wound irrigating agents used with commercially available wound dressings.

Increasing antibiotic resistance has led to an urgent need for new therapeutic approaches. Host defense peptides are known to be antimicrobial and have revealed broad immunomodulatory functions for both innate and adaptive immunity. This review will focus on the role of host defense peptides in infection and immune response and discuss its potential and limitations as a future therapeutical agent.

Background:Mucin 5AC (MUC5AC) belongs to the glycoprotein family of secreted gel-forming mucins and is physiologically expressed in some epithelial cells. Studies have shown that MUC5AC is also expressed in several cancer types suggesting a potential utility for the distinction of tumor types and subtypes. Methods:To systematically determine MUC5AC expression in normal and cancerous tissues, a tissue microarray containing 10 399 samples from 111 different tumor types and subtypes as well as 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. Results:MUC5AC was expressed in normal mucus-producing cells of various organs. At least weak MUC5AC positivity was seen in 44 of 111 (40%) tumor entities. Of these 44 tumor entities, 28 included also tumors with strong positivity. MUC5AC immunostaining was most commonly seen in esophageal adenocarcinoma (72%), colon adenoma (62%), ductal adenocarcinoma of the pancreas (64%), mucinous carcinoma of the ovary (46%), diffuse gastric adenocarcinoma (44%), pancreatic ampullary adenocarcinoma (41%), intestinal gastric adenocarcinoma (39%), and bronchioloalveolar carcinoma (33%). Clinically relevant tumors with complete or almost complete absence of MUC5AC staining included small cell carcinoma of the lung (0% of 17), clear cell renal cell carcinoma (0% of 507), papillary thyroid carcinoma (0% of 359), breast cancer (2% of 1097), prostate cancer (2% of 228), soft tissue tumors (0.1% of 968), and hematological neoplasias (0% of 111). Conclusion:The highly standardized analysis of a broad range of cancers identified a ranking order of tumors according to their relative prevalence of MUC5AC expression.

The temperature dependence of the oxidation kinetics of Fe²⁺ by O3 at pH 0–3 was studied by stopped-flow technique in the temperature range 5–40°C. Activation parameters of the reactions involved in formation and decay of the ferryl ion (iron(IV)), FeO²⁺ are determined. The reaction of Fe²⁺ + FeO²⁺ was found to branch into two channels forming iron(III)-dimer, Fe(OH)2Fe⁴⁺, and Fe³⁺. The yield of the dimer, Fe(OH)2Fe⁴⁺, increases with temperature on the expense of the Fe³⁺ yield. On the basis of the overall rate constant and relative yield of Fe(OH)2Fe⁴⁺ the activation energy is determined for both channels. The activation parameters of the hydrolysis of the ferryl ion and its reaction with H2O2 were also determined. © 1997 John Wiley & Sons, Inc. Int J Chem Kinet 29: 17–24, 1997.

The lifetime of the ferryl ion FeO²⁺, (minutes at pH below 1) is reduced by a factor of 50 at pH above 3. This is rationalized in terms of an acid-base equilibrium between two different hydrolytic forms of this species (Fe(OH)⁺⁺2 and Fe(OH)⁺3) with a pKa=2.0. The rate constants for reactions between FeO²⁺ and selected compounds found in cloud water were measured in acid solutions by stopped-flow technique. For inorganic reactants: kHNO2=1.1×10⁴ M⁻¹ s⁻¹, kNO2−≤10⁵ M⁻¹ s⁻¹, kCl−=1.0×10² M⁻¹ s^{− 1}, kHSO3−=2.5×10⁵ M⁻¹ s⁻¹, kSO2=4.5×10⁵ M⁻¹ s⁻¹, and kMn(II)=1.0×10⁴ M⁻¹ s⁻¹ were obtained and for the organic reactants: kHCOOH=160 M⁻¹ s⁻¹, kHCOO−=3×10⁵ M⁻¹, kCH3COOH=3.1 M⁻¹ s⁻¹, kCH2(OH)2=400 M⁻¹ s⁻¹, kCH3COCH3=16 M⁻¹ s⁻¹, kCH3CH20H=2.5×10³ M⁻¹ s⁻¹, kC6H5OH=4.0×10³ M⁻¹ s⁻¹, and kC6H5COOH=80 M⁻¹ s⁻¹ were obtained. A good correlation between log(k) and the standard one electron reduction potential (E°) indicates that the reactions of inorganic compounds proceed by electron transfer. The reaction mechanisms in case of organic compounds are very similar to the reactions of the OH radical, i.e., H-abstraction, and a fairly good correlation between log(k) and the bond dissociation energy BDE was obtained. Activation parameters were measured for the reaction of FeO²⁺ with HNO2 (Ea=34.5 kJ/mol); Mn²⁺ (E a=21.3 kJ/mol); HCOOH (Ea=22.3 kJ/mol); CH2 (OH)2 (Ea=44.5 kJ/mol); and C6H5 OH (Ea=28.1 kJ/mol). © 1998 John Wiley & Sons, Inc. Int J Chem Kinet 30: 215–221, 1998.

Twentieth‐century changes in Norwegian glaciers have been pronounced, but the different geometries and dynamics of the glaciers have caused different responses to similar climatic changes. Close to the Arctic Circle, all the glaciers of Svartisen, the largest ice‐covered area of northern Scandinavia, have retreated since the beginning of the century. However, several of the smaller glaciers which end at relatively high altitude have experienced both periods of advance and periods of retreat since the mid‐1960s. The mass balance of Engabreen, the largest of the West Svartisen glaciers, was positive in 21 of the 27 years to 1995–96. The sizes of most of the glaciers of the Okstindan area, 60 km south‐east of Svartisen, have also decreased throughout the twentieth century, but Corneliussens Bre, a small glacier at the eastern side of the massif, has been advancing since 1970. The areas supplying some of the southern glaciers of Okstindan have been reduced as a result of changes in ice thickness at high altitude. Studies of glacier change are aided by the use of digital terrain models (DTMs). Triangular irregular network DTMs of the surface and bed topography of the largest of the Okstindan glaciers, Austre Okstindbreen, have been used in studies of mass‐balance variations and changing surface flow patterns between 1976 and 1995.

Twentieth‐century changes in Norwegian glaciers have been pronounced, but the different geometries and dynamics of the glaciers have caused different responses to similar climatic changes. Close to the Arctic Circle, all the glaciers of Svartisen, the largest ice‐covered area of northern Scandinavia, have retreated since the beginning of the century. However, several of the smaller glaciers which end at relatively high altitude have experienced both periods of advance and periods of retreat since the mid‐1960s. The mass balance of Engabreen, the largest of the West Svartisen glaciers, was positive in 21 of the 27 years to 1995–96. The sizes of most of the glaciers of the Okstindan area, 60 km south‐east of Svartisen, have also decreased throughout the twentieth century, but Corneliussens Bre, a small glacier at the eastern side of the massif, has been advancing since 1970. The areas supplying some of the southern glaciers of Okstindan have been reduced as a result of changes in ice thickness at high altitude. Studies of glacier change are aided by the use of digital terrain models (DTMs). Triangular irregular network DTMs of the surface and bed topography of the largest of the Okstindan glaciers, Austre Okstindbreen, have been used in studies of mass‐balance variations and changing surface flow patterns between 1976 and 1995.

A remarkable contribution towards new leads for the fight against multiresistant microorganisms is discussed.