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Abstract This is the protocol for a Campbell systematic review. The objectives are as follows: Our proposed systematic review and meta‐analysis will integrate the evidence available from all sources to answer the following questions: (1) to what extent can information, behavioral and monetary interventions reduce energy consumption of households in residential buildings? (average treatment effect of interventions) (2) what is the relative effectiveness of interventions? (account for heterogeneity in treatment effects across and within studies) (3) how effective are combinations of different interventions?

Abstract The PEGASUS study is the first multicentric and prospective assessment of the safety of air travel flying in pulmonary hypertension (PH) (NCT03051763). Data of air travel from 60 patients with PH was available. No severe adverse events occurred. Nine patients self‐reported mild adverse events during flight (13%), while after landing, 12 patients reported events (20%). Solely one patient (2%) had an adverse event leading to medical consultation. In patients with PH and World Health Organization functional classes II and III, air travel was safe.

Abstract Objective Myasthenia gravis (MG) is an autoimmune disease leading to fatigable muscle weakness. Extra‐ocular and bulbar muscles are most commonly affected. We aimed to investigate whether facial weakness can be quantified automatically and used for diagnosis and disease monitoring. Methods In this cross‐sectional study, we analyzed video recordings of 70 MG patients and 69 healthy controls (HC) with two different methods. Facial weakness was first quantified with facial expression recognition software. Subsequently, a deep learning (DL) computer model was trained for the classification of diagnosis and disease severity using multiple cross‐validations on videos of 50 patients and 50 controls. Results were validated using unseen videos of 20 MG patients and 19 HC. Results Expression of anger (p = 0.026), fear (p = 0.003), and happiness (p

Abstract Aims Telemedical interventions in heart failure patients intend to avoid unfavourable, indication‐related events by an early, individualized care, which reacts to the current patients need. However, telemedical support is an expensive intervention, and usually only patients with high risk for unfavourable follow‐up events will be able to profit from it. Möckel et al. therefore adapted a new design which we call ‘prognostic‐efficacy‐combination design’. This design allows to define a biomarker cut‐off and to perform a randomized controlled trial (RCT) in a biomarker‐selected population within a single study. However, so far, it has not been evaluated if this double use of the control group for biomarker cut‐off definition and efficacy assessment within the RCT leads to a bias in treatment effect estimation. In this methodological research work, we therefore want to evaluate whether the ‘prognostic‐efficacy‐combination design’ leads to biased treatment effect estimates and also compare it to alternative designs. If there is a bias, we further want to analyse its magnitude under different parameter settings. Methods We perform a systematic Monte Carlo simulation study to investigate among others potential bias, root mean square error and sensitivity, and specificity as well as the total treatment effect estimate in various realistic trial scenarios that mimic and vary the true data characteristics of the published TIM‐HF2 Trial. In particular, we vary the event proportion, the sample size, the biomarker distribution, and the lower bound for the sensitivity. Results The results show that indeed the proposed design leads to some bias in the effect estimators, indicating an overestimation of the effect. However, this bias is relatively small in most scenarios. Conclusions The ‘prognostic‐efficacy‐combination design’ can generally be recommended for clinical applications due to its efficiency compared to two separate trials. We recommend a sufficiently large sample size depending on the trial scenario. Our simulation code can be adapted to explore suitable sample sizes for other settings.

Abstract Objective Activated astroglia is involved in the pathophysiology of neurodegenerative diseases and has also been described in animal models of spinal muscular atrophy (SMA). Given the urgent need of biomarkers for treatment monitoring of new RNA‐modifying and gene replacement therapies in SMA, we examined glial fibrillary acidic protein concentrations in cerebrospinal fluid (cGFAP) as a marker of astrogliosis in SMA. Methods 58 adult patients and 21 children with genetically confirmed 5q‐associated SMA from four German motor neuron disease specialist care centers and 30 age‐ and sex‐matched controls were prospectively included in this study. cGFAP was measured and correlated to motor performance and disease severity. Additionally, we compared cGFAP with neurofilament light chain concentrations in cerebrospinal fluid (cNfL). Results cGFAP concentrations did not differ from controls but showed higher levels in more severely affected patients after adjustment for patients' age. Normalized cNfL values were associated with disease severity. Within 14 months of nusinersen treatment, cGFAP concentrations did not change, while cNfL decreased significantly. Interpretation cGFAP is not an outstanding biomarker in SMA, but might support the hypothesis that glial activation is involved in SMA pathology. Unlike previously suggested, cNfL may be a promising biomarker also in adult patients with SMA, which should be subject to further investigations.

Abstract While the efficacy of the intracardiac defibrillators (ICDs) for primary prevention is not disputed, the relevant studies were carried out a long time ago. Most pertinent trials, including MADIT‐II, SCD‐Heft, and DEFINITE, recruited patients more than 20 years ago. Since then, improved therapeutic modalities including, in addition to cardiac resynchronization therapy, mineralocorticoid receptor antagonists, angiotensin receptor‐neprilysin inhibitors, and, most recently, inhibitors of sodium‐glucose cotransporter 2, have lowered present‐day rates of mortality and of sudden cardiac death. Thus, nowadays, ICD therapy may be less effective than previously reported, and not as beneficial as many people currently believe. However, criteria for ICD implantation remain very inclusive. The patient must (only) be symptomatic and have ejection fraction (EF) ≤ 35%. The choice of EF 35% is notable because the average EF in all large trials was much lower, and clinical benefit was mainly limited to EF ≤ 30%. This EF cut‐off value defines a substantial portion of potential ICD recipients. It seems therefore reasonable to limit ICD eligibility criteria in the EF range 30–35% to patients at highest risk only. We discuss and present some rational criteria to assist the clinician in improving risk stratification for preventive ICD implantation.

Abstract Aims This study aimed to investigate the left ventricular (LV) remodelling and long‐term prognosis of patients with new‐onset acute heart failure (HF) with reduced ejection fraction who were pharmacologically managed and survived until hospital discharge. We compared patients with ischaemic and non‐ischaemic aetiology. Methods and results This cohort study consisted of 111 patients admitted with new‐onset acute HF in the period 2008–2016 [62% non‐ischaemic aetiology, 48% supported by inotropes, vasopressors, or short‐term mechanical circulatory devices, and left ventricular ejection fraction (LVEF) at discharge 28% (interquartile range 22–34)]. LV dimensions, LVEF, and mitral valve regurgitation were used as markers for LV remodelling during up to 3 years of follow‐up. Both patients with non‐ischaemic and ischaemic HF had significant improvement in LVEF (P

Abstract Objective To determine whether serum creatine kinase activity (CK) and serum creatinine concentration (Crn) are prognostic and predictive biomarkers for disease severity, disease progression, and nusinersen treatment effects in adult patients with 5q‐associated spinal muscular atrophy (SMA). Methods Within this retrospective, multicenter observational study in 206 adult patients with SMA, we determined clinical subtypes (SMA types, ambulatory ability) and repeatedly measured CK and Crn and examined disease severity scores (Hammersmith Functional Motor Scale Expanded, Revised Upper Limb Module, and revised Amyotrophic Lateral Sclerosis Functional Rating Scale). Patients were followed under nusinersen treatment for 18 months. Results CK and Crn differed between clinical subtypes and correlated strongly with disease severity scores (e.g., for Hammersmith Functional Motor Scale Expanded: (CK) ρ = 0.786/ (Crn) ρ = 0.558). During the 18 months of nusinersen treatment, CK decreased (∆CK = −17.56%, p

Abstract Objective To quantify peripheral nerve lesions in symptomatic and asymptomatic hereditary transthyretin amyloidosis with polyneuropathy (ATTRv‐PNP) by analyzing the magnetization transfer ratio (MTR) of the sciatic nerve, and to test its potential as a novel biomarker for macromolecular changes. Methods Twenty‐five patients with symptomatic ATTRv‐PNP, 30 asymptomatic carriers of the mutant transthyretin gene (mutTTR), and 20 age‐/sex‐matched healthy controls prospectively underwent magnetization transfer contrast imaging at 3 Tesla. Two axial three‐dimensional gradient echo sequences with and without an off‐resonance saturation rapid frequency pulse were conducted at the right distal thigh. Sciatic nerve regions of interest were manually drawn on 10 consecutive axial slices in the images without off‐resonance saturation, and then transferred to the corresponding slices that were generated by the sequence with the off‐resonance saturation pulse. Subsequently, the MTR and cross‐sectional area (CSA) of the sciatic nerve were evaluated. Detailed neurologic and electrophysiologic examinations were conducted in all ATTRv‐PNP patients and mutTTR‐carriers. Results Sciatic nerve MTR and CSA reliably differentiated between ATTRv‐PNP, mutTTR‐carriers, and controls. MTR was lower in ATTRv‐PNP (26.4 ± 0.7; P

Abstract Objective The aim of this study was to assess the diagnostic value of point-of-care bedside ultrasound (PoCUS) as in usual clinical practice in suspected ankle and fifth metatarsal bone fractures, compared to the standard of radiographic imaging. Methods This prospective study included patients ≥17 years presenting to the Emergency Department with ankle trauma and positive Ottawa Ankle Rules. All patients underwent PoCUS of the ankle by a (resident) emergency physician, the images were assessed by an ultrasound expert. Both were blinded for the medical history and clinical findings of the patients. Radiography of the ankle followed, evaluated by a radiologist blinded from the PoCUS findings. Primary outcome measures were sensitivity and specificity of PoCUS. Results A total of 242 patients were included, with 35 (22%) clinically significant (non-avulsion) fractures observed with radiography. The sensitivity of PoCUS in detecting clinically significant fractures by all sonographers was 80.0% (95% Confidence Interval (CI) 63.0 to 91.6%), specificity 90.3% (95% CI 83.7 to 94.9%), positive predictive value 70.0% (95% CI 57.0 to 80.3%) and the negative predictive value 94.1% (95% CI 89.1 to 96.9%). The sensitivity of PoCUS in detecting clinically significant fractures by the expert was 82.8% (95% CI 66.3 to 93.4%), specificity 99.2% (95% CI 95.5 to 99.9%), positive predictive value 96.7% (95% CI 80.3 to 99.5%) and the negative predictive value 95.3% (95% CI 91.0 to 98.2%). Conclusion PoCUS combined with the OAR has a good diagnostic value in usual clinical practice in the assessment of suspected ankle and fifth metatarsal bone fractures compared to radiographic imaging. More experience with PoCUS will improve the diagnostic value. Trial registration Registered in the local Research Register, study number 201500597 .

Abstract Aggregation‐induced emission (AIE) is a phenomenon in which fluorescence is enhanced rather than quenched upon molecular assembly. AIE fluorogens (AIEgens) are flexible, conjugated systems that are limited in their dynamics when assembled, which improves their fluorescent properties. This intriguing feature has been incorporated in many different molecular assemblies and has been extended to nanoparticles composed of amphiphilic polymer building blocks. The integration of the fascinating AIE design principle with versatile polymer chemistry opens up new frontiers to approach and solve intrinsic obstacles of conventional fluorescent materials in nanoscience, including the aggregation‐caused quenching effect. Furthermore, this integration has drawn significant attention from the nanomedicine community, due to the additional advantages of nanoparticles comprising AIEgenic molecules, such as emission brightness and fluorescence stability. In this regard, a range of AIEgenic amphiphilic polymers have been developed, displaying enhanced emission in the self‐assembly/aggregated state. AIEgenic assemblies are regarded as attractive nanomaterials with inherent fluorescence, which display promising features in a biomedical context, for instance in biosensing, cell/tissue imaging and tracking, as well as (photo) therapeutics. In this review, we describe recent strategies for the design and synthesis of novel types of AIEgenic amphiphilic polymers via facile approaches including direct conjugation to natural/synthetic polymers, polymerization, post‐polymerization and supramolecular host−guest interactions. Their self‐assembly behavior and biomedical potential will be discussed.

Abstract Electrofishing is a well‐established and widely used method for surveying fish populations. Nonetheless, its effectiveness is impacted by numerous factors, including water chemistry, habitat type and fish species. Both physiological and behavioural responses make bottom‐dwelling ‘benthic’ fish which lack swim bladders (e.g. European bullhead Cottus gobio) particularly difficult to survey by electrofishing. We compare the performance and practicalities of electrofishing for benthic fish at a rocky northern English headwater stream with two sampling methods originally designed for crayfish surveys; the triple drawdown method which involves repeated dewatering of a site, and the Pritchard Trap method which involves sunken traps filled with natural substrate that samples a small, fixed (0.25 m2) area of river bed. Both the Pritchard trapping and triple drawdown methods provided similar high‐density population density estimates for bullhead which were at least 2.5–5 times higher than predicted from electrofishing derived sweep depletion curves. Electrofishing and the triple drawdown method are both resource‐intensive, requiring expensive equipment and a team of trained operatives. These approaches also pose a risk to fish and non‐target organisms. In contrast, Pritchard Traps provide a cost‐effective passive, low risk survey method requiring minimal training and only one operative. Pritchard traps, therefore, show particular promise for benthic fish surveying and monitoring.

Objective Investigation of the treatment of femur fractures and the type of femur fracture‐associated complications regarding timing of surgery and length of hospital stay. Methods In this retrospective cohort study, a total of 358 hip fractures were evaluated retrospectively from 1 January 2008 until 31 December 2010 at a level I trauma center in Germany. Inclusion criteria was age >18 years and a proximal femur fracture. Both sexes were evaluated. Mean age was 75.5 years, most patients were female (63.7%). Intervention was the operative treatment of proximal femur fracture. Outcome parameters were time until surgery, complications, reoperations, mortality, and length of hospital stay. Results Among the proximal femur fractures (n = 358), 46.6% were pertrochanteric, 11.2% subtrochanteric, and 42.2% femoral neck fractures. Operation upon hip fractures was managed regularly within 24 hours of injury (73%; mean for femoral neck: 28.3 hrs.; mean for pertrochanteric fractures: 21.4 hrs.; mean for subtrochanteric fractures: 19.5 hrs.). Delayed treatment, as well as implantation of hip total endoprosthesis (TEP), increased the overall length of hospital stay (15.4 vs 17.6 days; 18.1 vs 15.8 days). Accordingly, surgical procedures performed within 24 hours of injury resulted in a shorter hospital residence. Longest delay of operation was measured for hip fractures (28.3 hrs.). In 351 patients, secondary injuries were detected in 94 individuals (26%), with fractures being the most common secondary injury (n = 40). We recorded postoperative complications of nonsurgical and surgical origin, and 33.6% of our patient cohort displayed complications. Complications were distributed among 118 patients. There was no significant difference in complications regarding the time of operation, with most nonsurgical and surgical complications appearing within 24 hours after operation (n = 110 vs n = 31). Nonsurgical complications, such as anemia (n = 49) and electrolyte imbalances (n = 30), were observed more frequently than surgical complications (n = 107 vs n = 34); however, these complications were reduced by delay in surgery (82.0% in 6–24 hrs. vs 74.2% in ≥24 hrs.). Anticoagulant therapy and age did not affect postoperative complications. The hospital mortality of patients was 6.2%. Follow‐up was restrained to ambulatory visits in the clinic. Conclusions Surgical management of hip fractures performed within 24 hours of injury minimizes hospital stay. We did not detect significant differences in the spectrum or number of complications regarding delay of surgery. Surgical complications mainly occur with rapid primary care, and medical complications can be reduced by more intensive preparation of patient and operation procedures.

Abstract As crayfish invasions continue to threaten native freshwater biota, a detailed understanding of crayfish distribution and population structure becomes imperative. Nonetheless, most current survey methods provide inadequate demographic data. The quantitative ‘Triple Drawdown’ (TDD) dewatering method has highlighted the importance of such data, yet practical constraints prevent its large‐scale application. Here, we introduce the ‘Pritchard Trap’, a novel passive sampling method that reliably generates quantitative crayfish population data while requiring substantially lower sampling effort than TDDs. This quadrat‐style sampler was extensively tested in headwater streams of North Yorkshire, England, along an invasion gradient for signal crayfish (Pacifastacus leniusculus) from well‐established sites to mixed populations of signal crayfish and native white‐clawed crayfish (Austropotamobius pallipes). The Pritchard Trap was trialled over several time intervals to determine the minimum required trap deployment time. TDDs at the same sites allowed for a robust evaluation of Pritchard Trap sampling accuracy in representing crayfish densities and population structure. The Pritchard Trap successfully sampled both invasive and native crayfish (8–42 mm carapace length). A minimum passive deployment time of 4 days was required. At low crayfish densities (0.5 individuals m−2), increased trapping effort was necessary to achieve accurate population density and size class distribution estimates. The Pritchard Trap required substantially less sampling effort (working hours) and resources than the TDD, whilst also posing less risk to non‐target species. The Pritchard Trap, for the first time, affords logistically simple, truly quantitative investigations of crayfish population demographics for headwater systems. It could be integrated into crayfish research and management, for example to explore density‐dependent ecological impacts of invasive crayfish and their management responses or to monitor populations and recruitment in native crayfish conservation initiatives.

Abstract Purpose Meniscus allograft transplantation (MAT) is a possible treatment for patients suffering with pain after meniscectomy. Here, peracetic acid (PAA) sterilised meniscus transplants were investigated on whether they would provide an adequate alternative to fresh‐frozen transplants in their viscoelastic and mechanical properties. Methods In this analysis, 31 menisci donors (26 male and 5 female) were included. The average donor age was 49.87 years, ranging from 32 to 65 years. Menisci of matched pairs of knees underwent chemical sterilisation while counterparts were left fresh‐frozen. Stiffness and load to failure were determined via suture retention. Further menisci were analysed while attached to the tibial bone block using a novel test device to mimic physiological load distribution. Meniscus relaxation, stiffness and failure loads were determined. Histology and biphasic properties of the menisci were examined and results were analysed using paired t‐tests. Results A novel custom built test device allowed the application of physiological loads for suture retention testing and revealed no significant differences between PAA sterilised (14.85 ± 4.46 N/mm, 50.49 ± 17.01 N) and fresh‐frozen (18.26 ± 4.46 N/mm, 59.49 ± 21.07 N) regarding stiffness and failure load, respectively. Furthermore, initial 200 N loading showed significantly higher strain in sterilised menisci (18.87 ± 1.56) compared to fresh frozen (13.81 ± 1.04). Load relaxation experiments demonstrated significantly lower relaxation for sterilised menisci (77.71 ± 1.62) compared to fresh‐frozen (89.11 ± 1.00, p‐value

Abstract The volume of published academic research is growing rapidly and this new era of “big literature” poses new challenges to evidence synthesis, pushing traditional, manual methods of evidence synthesis to their limits. New technology developments, including machine learning, are likely to provide solutions to the problem of information overload and allow scaling of systematic maps to large and even vast literatures. In this paper, we outline how systematic maps lend themselves well to automation and computer‐assistance. We believe that it is a major priority to consolidate efforts to develop and validate efficient, rigorous and robust applications of these novel technologies, ensuring the challenges of big literature do not prevent the future production of systematic maps.

Abstract Background Matrix metallopeptidase 20 (MMP20) is an evolutionarily conserved protease that is essential for processing enamel matrix proteins during dental enamel formation. MMP20 mutations cause human autosomal recessive pigmented hypomaturation‐type amelogenesis imperfecta (AI2A2; OMIM #612529). MMP20 is expressed in both odontoblasts and ameloblasts, but its function during dentinogenesis is unclear. Methods We characterized 10 AI kindreds with MMP20 defects, characterized human third molars and/or Mmp20−/− mice by histology, Backscattered Scanning Electron Microscopy (bSEM), µCT, and nanohardness testing. Results We identified six novel MMP20 disease‐causing mutations. Four pathogenic variants were associated with exons encoding the MMP20 hemopexin‐like (PEX) domain, suggesting a necessary regulatory function. Mutant human enamel hardness was softest (13% of normal) midway between the dentinoenamel junction (DEJ) and the enamel surface. bSEM and µCT analyses of the third molars revealed reduced mineral density in both enamel and dentin. Dentin close to the DEJ showed an average hardness number 62%–69% of control. Characterization of Mmp20−/− mouse dentin revealed a significant reduction in dentin thickness and mineral density and a transient increase in predentin thickness, indicating disturbances in dentin matrix secretion and mineralization. Conclusion These results expand the spectrum of MMP20 disease‐causing mutations and provide the first evidence for MMP20 function during dentin formation.

Pulmonary hypertension is frequently underdiagnosed, and referral is delayed with subsequent impact on outcomes. During the SARS-CoV-2 pandemic, restrictions on daily life and changes in hospitals' daily routine care were introduced in Germany. This multi-centre study provides evidence for a negative influence of these restrictions on patient care in pulmonary hypertension expert referral centres.

Abstract Nanoparticle‐based drug delivery systems with low side effects and enhanced efficacy hold great potential in the treatment of various malignancies, in particular cancer; however, they are still challenging to attain. Herein, an anticancer drug delivery system based on a cisplatin (CDDP) containing nanogel, functionalized with photothermal gold nanorods (GNRs) which are electrostatically decorated with doxorubicin (DOX) is reported. The nanoparticles are formed via the crosslinking reaction of hyaluronic acid with the ancillary anticarcinogen CDDP in the presence of DOX‐decorated GNRs. The nanogel is furthermore cloaked with a cancer cell membrane, and the resulting biomimetic nanocarrier (4T1‐HANG‐GNR‐DC) shows efficient accumulation by homologous tumor targeting and possesses long‐time retention in the tumor microenvironment. Upon near‐infrared (NIR) laser irradiation, in situ photothermal therapy is conducted which further induces hyperthermia‐triggered on‐demand drug release from the nanogel reservoir to achieve a synergistic photothermal/chemo‐therapy. The as‐developed biomimetic nanocarriers, with their dual‐drug delivery features, homotypic tumor targeting and synergetic photothermal/chemo‐therapy, show much promise as a potential platform for cancer treatment.

Abstract Most of Earth's biodiversity is found in 36 biodiversity hotspots, yet less than 10% natural intact vegetation remains. We calculated models projecting the future state of most of these hotspots for the year 2050, based on future climatic and agroeconomic pressure. Our models project an increasing demand for agricultural land resulting in the conversion of >50% of remaining natural intact vegetation in about one third of all hotspots, and in 2–6 hotspots resulting from climatic pressure. This confirms that, in the short term, habitat loss is of greater concern than climate change for hotspots and their biodiversity. Hotspots are most severely threatened in tropical Africa and parts of Asia, where demographic pressure and the demand for agricultural land is highest. The speed and magnitude of pristine habitat loss is, according to our models, much greater than previously shown when combining both scenarios on future climatic and agroeconomic pressure.

Abstract Background ENAM mutations cause autosomal dominant or recessive amelogenesis imperfecta (AI) and show a dose effect: enamel malformations are more severe or only penetrant when both ENAM alleles are defective. Methods Whole exome sequences of recruited AI probands were initially screened for mutations in known AI candidate genes. Sanger sequencing was used to confirm sequence variations and their segregation with the disease phenotype. The co‐occurrence of ENAM and LAMA3 mutations in one family raised the possibility of digenic inheritance. Enamel formed in Enam+/+Ambn+/+, Enam+/−, Ambn+/−, and Enam+/−Ambn+/− mice was characterized by dissection and backscattered scanning electron microscopy (bSEM). Results ENAM mutations segregating with AI in five families were identified. Two novel ENAM frameshift mutations were identified. A single‐nucleotide duplication (c.395dupA/p.Pro133Alafs*13) replaced amino acids 133‐1142 with a 12 amino acid (ATTKAAFEAAIT*) sequence, and a single‐nucleotide deletion (c.2763delT/p.Asp921Glufs*32) replaced amino acids 921‐1142 with 31 amino acids (ESSPQQASYQAKETAQRRGKAKTLLEMMCPR*). Three families were heterozygous for a previously reported single‐nucleotide ENAM deletion (c.588+1delG/p.Asn197Ilefs*81). One of these families also harbored a heterozygous LAMA3 mutation (c.1559G>A/p.Cys520Tyr) that cosegregated with both the AI phenotype and the ENAM mutation. In mice, Ambn+/− maxillary incisors were normal. Ambn+/− molars were also normal, except for minor surface roughness. Ambn+/− mandibular incisors were sometimes chalky and showed minor chipping. Enam+/− incisor enamel was thinner than normal with ectopic mineral deposited laterally. Enam+/− molars were sometimes chalky and rough surfaced. Enam+/−Ambn+/− enamel was thin and rough, in part due to ectopic mineralization, but also underwent accelerated attrition. Conclusion Novel ENAM mutations causing AI were identified, raising to 22 the number of ENAM variations known to cause AI. The severity of the enamel phenotype in Enam+/−Ambn+/− double heterozygous mice is caused by composite digenic effects. Digenic inheritance should be explored as a cause of AI in humans.

Abstract Background Ameloblastin (AMBN) is a secreted matrix protein that is critical for the formation of dental enamel and is enamel‐specific with respect to its essential functions. Biallelic AMBN defects cause non‐syndromic autosomal recessive amelogenesis imperfecta. Homozygous Ambn mutant mice expressing an internally truncated AMBN protein deposit only a soft mineral crust on the surface of dentin. Methods We characterized a family with hypoplastic amelogenesis imperfecta caused by AMBN compound heterozygous mutations (c.1061T>C; p.Leu354Pro/ c.1340C>T; p.Pro447Leu). We generated and characterized Ambn knockout/NLS‐lacZ (AmbnlacZ/lacZ) knockin mice. Results No AMBN protein was detected using immunohistochemistry in null mice. ß‐galactosidase activity was specific for ameloblasts in incisors and molars, and islands of cells along developing molar roots. AmbnlacZ/lacZ 7‐week incisors and unerupted (D14) first molars showed extreme enamel surface roughness. No abnormalities were observed in dentin mineralization or in nondental tissues. Ameloblasts in the AmbnlacZ/lacZ mice were unable to initiate appositional growth and started to degenerate and deposit ectopic mineral. No layer of initial enamel ribbons formed in the AmbnlacZ/lacZ mice, but pockets of amelogenin accumulated on the dentin surface along the ameloblast distal membrane and within the enamel organ epithelia (EOE). NLS‐lacZ signal was positive in the epididymis and nasal epithelium, but negative in ovary, oviduct, uterus, prostate, seminal vesicles, testis, submandibular salivary gland, kidney, liver, bladder, and bone, even after 15 hr of incubation with X‐gal. Conclusions Ameloblastin is critical for the initiation of enamel ribbon formation, and its absence results in pathological mineralization within the enamel organ epithelia.

Abstract Distinguishing tumors from normal brain cells is important but challenging in glioma surgery due to the lack of clear interfaces between the two. The ability of label‐free third harmonic generation (THG) microscopy in combination with automated image analysis to quantitatively detect glioma infiltration in fresh, unprocessed tissue in real time is assessed. The THG images reveal increased cellularity in grades II–IV glioma samples from 23 patients, as confirmed by subsequent hematoxylin and eosin histology. An automated image quantification workflow is presented for quantitative assessment of the imaged cellularity as a reflection of the degree of glioma invasion. The cellularity is validated in three ways: 1) Quantitative comparison of THG imaging with fluorescence microscopy of nucleus‐stained samples demonstrates that THG reflects the true tissue cellularity. 2) Thresholding of THG cellularity differentiates normal brain from glioma infiltration, with 96.6% sensitivity and 95.5% specificity, in nearly perfect (93%) agreement with pathologists. 3) In one patient, a good correlation between THG cellularity and preoperative magnetic resonance and positron emission tomography imaging is demonstrated. In conclusion, quantitative real‐time THG microscopy accurately assesses glioma infiltration in ex vivo human brain samples, and therefore holds strong potential for improving the accuracy of surgical resection.

Abstract Background Truncation FAM83H mutations cause human autosomal dominant hypocalcified amelogenesis imperfecta (ADHCAI), an inherited disorder characterized by severe hardness defects in dental enamel. No enamel defects were observed in Fam83h null mice suggesting that Fam83h truncation mice would better replicate human mutations. Methods We generated and characterized a mouse model (Fam83hTr/Tr) expressing a truncated FAM83H protein (amino acids 1–296), which recapitulated the ADHCAI‐causing human FAM83H p.Tyr297* mutation. Results Day 14 and 7‐week Fam83hTr/Tr molars exhibited rough enamel surfaces and slender cusps resulting from hypoplastic enamel defects. The lateral third of the Fam83hTr/Tr incisor enamel layer was thinner, with surface roughness and altered enamel rod orientation, suggesting disturbed enamel matrix secretion. Regular electron density in mandibular incisor enamel indicated normal enamel maturation. Only mildly increased posteruption attrition of Fam83hTr/Tr molar enamel was observed at 7‐weeks. Histologically, the Fam83hTr/Tr enamel organ, including ameloblasts, and enamel matrices at sequential stages of amelogenesis exhibited comparable morphology without overt abnormalities, except irregular and less evident ameloblast Tomes' processes in specific areas. Conclusions Considering Fam83h−/− mice showed no enamel phenotype, while Fam83hTr/Tr (p.Tyr297*) mice displayed obvious enamel malformations, we conclude that FAM83H truncation mutations causing ADHCAI in humans disturb amelogenesis through a neomorphic mechanism, rather than haploinsufficiency.

Abstract The PREDICTS project—Projecting Responses of Ecological Diversity In Changing Terrestrial Systems (https://www.nhm.ac.uk/our-science/our-work/biodiversity/predicts.html)—has collated from published studies a large, reasonably representative database of comparable samples of biodiversity from multiple sites that differ in the nature or intensity of human impacts relating to land use. We have used this evidence base to develop global and regional statistical models of how local biodiversity responds to these measures. We describe and make freely available this 2016 release of the database, containing more than 3.2 million records sampled at over 26,000 locations and representing over 47,000 species. We outline how the database can help in answering a range of questions in ecology and conservation biology. To our knowledge, this is the largest and most geographically and taxonomically representative database of spatial comparisons of biodiversity that has been collated to date; it will be useful to researchers and international efforts wishing to model and understand the global status of biodiversity.