Dual truncation of tau by caspase-2 accelerates its CHIP-mediated degradation.

Intraneuronal aggregates of the microtubule binding protein Tau are a hallmark of different neurodegenerative diseases including Alzheimer's disease (AD). In these aggregates, Tau is modified by posttranslational modifications such as phosphorylation as well as by proteolytic cleavage. Here we identify a novel Tau cleavage site at aspartate 65 (D65) that is specific for caspase-2. In addition, we show that the previously described cleavage site at D421 is also efficiently processed by caspase-2, and both sites are cleaved in human brain samples. Caspase-2-generated Tau fragments show increased aggregation potential in vitro, but do not accumulate in vivo after AAV-mediated overexpression in mouse hippocampus. Interestingly, we observe that steady-state protein levels of caspase-2 generated Tau fragments are low in our in vivo model despite strong RNA expression, suggesting efficient clearance. Consistent with this hypothesis, we find that caspase-2 cleavage significantly improves the recognition of Tau by the ubiquitin E3 ligase CHIP, leading to increased ubiquitination and faster degradation of Tau fragments. Taken together our data thus suggest that CHIP-induced ubiquitination is of particular importance for the clearance of caspase-2 generated Tau fragments in vitro and in vivo.

TOPICAL CARBONIC ANHYDRASE INHIBITORS IN THE LONG-TERM TREATMENT OF JUVENILE X-LINKED RETINOSCHISIS.

PURPOSE: To describe the response to long-term topical dorzolamide treatment in patients with juvenile X-linked retinoschisis and cystic-like foveal lesions. METHODS: This was a retrospective interventional case series that included 18 eyes of 10 patients with genetically confirmed juvenile X-linked retinoschisis examined at the Cleveland Clinic Cole Eye Institute, a tertiary referral center, between 2005 and 2021. Patients were treated with topical 2% dorzolamide two to three times daily in both eyes. Two eyes were excluded because of retinal detachment. Primary outcome measures were logarithm of minimum angle of resolution visual acuity and optical coherence tomography based central subfield thickness. RESULTS: The mean follow-up was 8.38 years (SD, 3.41 years). The mean baseline and final central subfield thickness was 429.88 µ m (SD, 143.36 µ m) and 372.28 µ m, respectively (SD, 147.13 µ m, P = 0.10). The mean baseline and final logarithm of minimum angle of resolution visual acuity was 0.45 (SD, 0.17) and 0.34, respectively (SD, 0.22, P < 0.01). None of the patients experienced any side effects from topical dorzolamide. CONCLUSION: The study data support previous reports of improved visual acuity in X-linked retinoschisis patients on topical dorzolamide treatment. This is the longest follow-up for a series of juvenile X-linked retinoschisis patients treated with a topical carbonic anhydrase inhibitor to date. A large, prospective, randomized clinical trial is needed to provide stronger evidence regarding the efficacy of topical carbonic anhydrase inhibitors in juvenile X-linked retinoschisis.

Loss of stromal JUNB does not affect tumor growth and angiogenesis.

The transcription factor AP-1 subunit JUNB has been shown to play a pivotal role in angiogenesis. It positively controls angiogenesis by regulating Vegfa as well as the transcriptional regulator Cbfb and its target Mmp13. In line with these findings, it has been demonstrated that tumor cell-derived JUNB promotes tumor growth and angiogenesis. In contrast to JUNB's function in tumor cells, the role of host-derived stromal JUNB has not been elucidated so far. Here, we show that ablation of Junb in stromal cells including endothelial cells (ECs), vascular smooth muscle cells (SMCs) and fibroblasts does not affect tumor growth in two different syngeneic mouse models, the B16-F1 melanoma and the Lewis lung carcinoma model. In-depth analyses of the tumors revealed that tumor angiogenesis remains unaffected as assessed by measurements of the microvascular density and relative blood volume in the tumor. Furthermore, we could show that the maturation status of the tumor vasculature, analyzed by the SMC marker expression, α-smooth muscle actin and Desmin, as well as the attachment of pericytes to the endothelium, is not changed upon ablation of Junb. Taken together, these results indicate that the pro-angiogenic functions of stromal JUNB are well compensated with regard to tumor angiogenesis and tumor growth.

A case report of retinal dystrophy in patients with PACS1 syndrome.

PACS1 syndrome, also referred to as Schuurs-Hoeijmakers syndrome, is a multisystemic developmental disorder caused by a specific pathogenic variant in the PACS1 (phosphofurin acidic cluster sorting protein 1) gene. Ocular findings in PACS1 syndrome are known to include iris, retina, optic nerve coloboma, myopia, nystagmus, and strabismus. Here, we present the cases of two patients referred to the University of Wisconsin-Madison Department of Ophthalmology and Visual Sciences for ocular evaluation. The first patient is a 14-month-old female who, at 3 months of age, was found to have a depressed rod and cone response on electroretinogram (ERG), consistent with possible retinal dystrophy (RD). This feature has not been previously described in PACS1 syndrome and joins a growing list of calls for expanding the PACS1 phenotype. The second case illustrates a 5-year-old male referred for ocular screening after diagnosing PACS1 syndrome and underwent ERG without abnormal findings. These cases demonstrate the significant variability in the ophthalmic presentation of PACS1 syndrome and the need for early screening. These novel findings may have implications in understanding the mechanism of the PACS1 protein and its role in retinal ciliary phototransduction in photoreceptors.

MFRP variant results in nanophthalmos, retinitis pigmentosa, variability in foveal avascular zone.

BACKGROUND: Membrane frizzled-related protein (MFRP) plays a critical role in ocular development. MFRP mutations are known to cause nanophthalmos and, in some cases, retinitis pigmentosa, foveoschisis, and/or optic nerve head (ONH) drusen. The broad clinical spectrum of MFRP mutations necessitates further investigation of specific genotype-phenotype relationships. MATERIALS AND METHODS: We reviewed ophthalmologic and genetic medical records of two affected siblings and one unaffected sibling. RESULTS: Genetic testing revealed variants MFRP c.855T>A, p.(Cys285*) and MFRP c.1235T>C, p.(Leu412Pro) in trans in the two affected siblings. In both cases, photopic and scotopic responses were markedly reduced on electroretinogram (ERG), with greater decrease in scotopic function. Optical coherence tomography for both siblings revealed non-cystoid thickening. Blunted foveal reflexes were also observed in both siblings. Notably, foveal avascular zone abnormalities were seen on fundus autofluorescence in only one affected sibling. CONCLUSIONS: MFRP-related ocular disease may be underrecognized due to its presentation with high hyperopia and possibly subtle retinal findings. Presence of variants MFRP c.855T>A, p.(Cys285*) and MFRP c.1235T>C, p.(Leu412Pro) in trans resulted in nanophthalmos and retinitis pigmentosa without associated foveoschisis or ONH drusen in our patients, consistent with the incomplete phenotype previously described in Neri et al. Abnormalities in the foveal avascular zone have been noted in other case studies and were inconsistently associated with the variants described here, representing a potential area for future investigation.

Retained avidity despite reduced cross-binding and cross-neutralizing antibody levels to Omicron after SARS-COV-2 wild-type infection or mRNA double vaccination.

Introduction: The rapid evolution of SARS-CoV-2 has posed a challenge to long-lasting immunity against the novel virus. Apart from neutralizing function, binding antibodies induced by vaccination or infection play an important role in containing the infection. Methods: To determine the proportion of wild-type (WT)-generated antibodies recognizant of more recent variants, plasma samples from either SARS-CoV-2 WT-infected (n = 336) or double-mRNA (Comirnaty)-vaccinated individuals (n = 354, age and sex matched to the convalescent group) were analyzed for binding antibody capacity against the S1 protein of the BA.1 omicron variant. Results: Overall, 38.59% (95% CI, 37.01- 40.20) of WT-generated antibodies recognized Omicron BA.1 S1 protein [28.83% (95% CI, 26.73-30.91) after infection and 43.46% (95% CI, 41.61-45.31) after vaccination; p < 0.001]. Although the proportion of WT-generated binding and neutralizing antibodies also binding to BA.1 is substantially reduced, the avidity of the remaining antibodies against the Omicron variant was non-inferior to that of the ancestral virus: Omicron: 39.7% (95% CI: 38.1-41.3) as compared to the avidity to WT: 27.0% (95% CI, 25.5-28.4), respectively (p < 0.001). Furthermore, we noticed a modestly yet statistically significant higher avidity toward the Omicron epitopes among the vaccinated group (42.2%; 95% CI, 40.51-43.94) as compared to the convalescent counterparts (36.4%; 95% CI, 33.42-38.76) (p = 0.003), even after adjusting for antibody concentration. Discussion: Our results suggest that an aspect of functional immunity against the novel strain was considerably retained after WT contact, speculatively counteracting the impact of immune evasion toward neutralization of the strain. Higher antibody levels and cross-binding capacity among vaccinated individuals suggest an advantage of repeated exposure in generating robust immunity.

CMTM6 shapes antitumor T cell response through modulating protein expression of CD58 and PD-L1.

The dysregulated expression of immune checkpoint molecules enables cancer cells to evade immune destruction. While blockade of inhibitory immune checkpoints like PD-L1 forms the basis of current cancer immunotherapies, a deficiency in costimulatory signals can render these therapies futile. CD58, a costimulatory ligand, plays a crucial role in antitumor immune responses, but the mechanisms controlling its expression remain unclear. Using two systematic approaches, we reveal that CMTM6 positively regulates CD58 expression. Notably, CMTM6 interacts with both CD58 and PD-L1, maintaining the expression of these two immune checkpoint ligands with opposing functions. Functionally, the presence of CMTM6 and CD58 on tumor cells significantly affects T cell-tumor interactions and response to PD-L1-PD-1 blockade. Collectively, these findings provide fundamental insights into CD58 regulation, uncover a shared regulator of stimulatory and inhibitory immune checkpoints, and highlight the importance of tumor-intrinsic CMTM6 and CD58 expression in antitumor immune responses.

A comprehensive structure-function comparison of hepatitis C virus strain JFH1 and J6 polymerases reveals a key residue stimulating replication in cell culture across genotypes.

The hepatitis C virus (HCV) genotype 2a isolate JFH1 represents the only cloned HCV wild-type sequence capable of efficient replication in cell culture as well as in vivo. Previous reports have pointed to NS5B, the viral RNA-dependent RNA polymerase (RdRp), as a major determinant for efficient replication of this isolate. To understand the contribution of the JFH1 NS5B gene at the molecular level, we aimed at conferring JFH1 properties to NS5B from the closely related J6 isolate. We created intragenotypic chimeras in the NS5B regions of JFH1 and J6 and compared replication efficiency in cell culture and RdRp activity of the purified proteins in vitro, revealing more than three independent mechanisms conferring the role of JFH1 NS5B in efficient RNA replication. Most critical was residue I405 in the thumb domain of the polymerase, which strongly stimulated replication in cell culture by enhancing overall de novo RNA synthesis. A structural comparison of JFH1 and J6 at high resolution indicated a clear correlation of a closed-thumb conformation of the RdRp and the efficiency of the enzyme at de novo RNA synthesis, in accordance with the proposal that I405 enhances de novo initiation. In addition, we identified several residues enhancing replication independent of RdRp activity in vitro. The functional properties of JFH1 NS5B could be restored by a few single-nucleotide substitutions to the J6 isolate. Finally, we were able to enhance the replication efficiency of a genotype 1b isolate with the I405 mutation, indicating that this mechanism of action is conserved across genotypes.

Intraventricular cooling during CSF infusion studies.

We implemented ventricular infusion studies on 33 patients suspected of idiopathic normal pressure hydrocephalus (iNPH), benign intracranial hypertension (BIH) or occlusive hydrocephalus (HOC) in order to confirm shunt indications. The initial scope was to study O(2) supply during infusion tests to exclude further violation of already vulnerable brains during ICP elevation. Intraventricular infusion was performed via ventricle catheters with the ICP tip sensor, while brain tissue oxygenation was measured with intraparenchymal Raumedic PTO probes. In 15 out of 23 (65%; 8 NPH, 2BIH, 5 HOC), pO(2) increased constantly (average 140%), while brain temperature decreased (range: 0.2-4.5°C) during the infusion studies. In another six patients, O(2) values remained largely stable during the infusion studies (4NPH, 1BIH, 1HOC). Cerebral deoxygenation during infusion tests occurred only in two patients (1NPH, 1HOC).Overall cerebral oxygenation and temperature inversely correlated well with some temporary delay regarding oxygenation state as a consequence of cerebral temperature. Probably, this effect is a consequence of reduced cerebral metabolism caused by local cooling. We hypothesise that such cooling is mediated via the large basal arteries and suggest that such a pathophysiology, ICP-controlled local cooling, might offer a new option for brain protection (e.g. in an ICP crisis).

Subdural or intraparenchymal placement of long-term telemetric intracranial pressure measurement devices?

We established a CE-certified telemetric device to measure intracranial pressure (ICP) noninvasively. To evaluate whether subdural or intraparenchymal insertion of such devices should be preferred, we implanted these telemetric ICP measurement devices (Raumedic, Rautel) in both locations. The study was performed in nine minipigs. The telemetric data were validated every 3 months using conventional intraparenchymal ICP measurement probes.The intraparenchymal telemetric device failed in one animal 12 months after insertion. Computed tomography (CT) revealed first hints for failure: Despite the implantation in adult animals, the skull dimensions seemingly increased after implantation, and the sensor tip was dislocated on the tabula interna level. This finding could also be verified by histopathological examination which would explain the reason for mismeasurement. The subdural catheter failed after 9 months. CT and histopathological examination revealed a bony encapsulation of a large catheter part, which had been located correctly initially. We propose that chronic pulsatile stress on the device was the underlying reason for this phenomenon, comparable to that in meningeal arteries.In some of the other animals, failure of subdural catheters could be detected. Histopathological examinations in these cases are still pending. Nevertheless, we assume similar underlying reasons for failure in these subdural probes.In conclusion, we favour intraparenchymal placement of telemetric ICP measurement devices.

Pathophysiology of brainstem lesions due to overdrainage.

Overdrainage in hydrocephalus therapy is a common shunt complication responsible for many different side effects. Especially an association with an impairment of upper brainstem structures causing symptoms of a dorsal midbrain syndrome (DMS) has already been described. Yet apart from these known mesencephalic lesions, we found several more brainstem signs and symptoms resulting from overdrainage. Parinaud's syndrome was diagnosed in all six patients examined; moreover, parkinsonism, memory disturbances, fluctuations in the level of consciousness, and hypothalamic dysfunctions could be detected in five of six patients. In addition hypersalivation combined with peripheral facial nerve palsy and blepharospasm occurred in two patients each, respectively. We postulate an upward herniation of the midbrain into the tentorial notch causing a secondary aqueductal stenosis as causal. An obstructed Sylvian aqueduct and the occurrence of shunt failure can lead to a bulging or enlargement of the third ventricle resulting in diencephalic lesions. If combined with fourth ventricle outlet occlusion, secondary aqueductal stenosis aggravates the situation with a fourth ventricle entrapment. Symptomatology and proposed pathophysiology are presented.

Dynamics of cerebrospinal fluid flow in slit ventricle syndrome.

INTRODUCTION: Although slit ventricle syndrome (SVS) is identified as a serious complication in shunt-treated hydrocephalus, cerebral spinal fluid (CSF) flow via external ventricular drainage (EVD) or shunts in SVS have not been studied up to now. MATERIAL AND METHODS: A new apparatus (LiquoGuard(®); Möller-Medical, Fulda, Germany) was used for EVD in a child with SVS. The LiquoGuard actively controls CSF drainage, based on intracranial pressure (ICP). RESULTS: To achieve well-tolerated clinical conditions, an ICP level of 4 mmHg was necessary; realizable by drainage rates between 0 and 35 mL/h. Drainage rate variations typically occurred with repetitive time intervals of 2 h causing a "saw tooth" shaped CSF flow pattern throughout 24 h. DISCUSSION: SVS seems to be characterized largely by quickly varying CSF drainage demands. Whether this is a general phenomenon or just true for this case has still to be studied and needs further clarification.

Telemetric ICP measurement with the first CE-approved device: data from animal experiments and initial clinical experiences.

The objective was to evaluate the qualification of the new telemetric intracranial pressure (ICP) measurement (t-ICP) device Raumedic(®) NEUROVENT P-Tel and S-Tel. The proof of concept was examined in a pilot animal study measuring intraperitoneal pressure with a telemetric and a conventional ICP measurement probe at five rates for 1 h each. Moderate external pressure load allowed measuring values between 0 and 40 mmHg. To estimate long-term performance 18 t-ICP devices were implanted subdurally or intraparenchymally into minipigs. Reference measurements were performed regularly using conventional ICP probes. From the short-term as well as from the long-term perspective t-ICP proved to have excellent dynamic ICP signal components perception (e.g. pulse amplitude). Some zero drift of static ICP was found, ranging between 5 and 8 mmHg. While all telemetric, intraparenchymal probes kept their functionality throughout the follow-up, 33% of the subdurals failed for reasons detailed in another paper. Raumedic's NEUROVENT(®) P-Tel/S-Tel proved to provide reliable data over periods of up to 18 months. Minor zero drift can be well tolerated as the dynamic ICP signal is measured with excellent stability. Clinicians should focus more on such ICP dynamic signal information than on static ICP when using the device over longer follow-up periods.

Frontal and temporal horn ratio: a valid and reliable index to determine ventricular size in paediatric hydrocephalus patients?

Because there is currently no sufficient and prevalent parameter for estimating ventricular size in paediatric hydrocephalus patients by using cranial ultrasound, a new measurement index, called the "frontal and temporal horn ratio", is presented in this study. The advantage of the new quotient is that it can be detected in easily obtainable coronal ultrasound planes. A retrospective analysis of 149 MRIs of young hydrocephalus patients proved the new index to be a promising parameter of ventricular size assessment. Statistical comparison between the "frontal and temporal horn ratio" and the already validated "frontal and occipital horn ratio" revealed a strong and linear correlation between the two quotients. Current research is now evaluating the reliability of the new index in the context of an ultrasound study; first results indicate similar positive findings.

Microstructural alterations of silicone catheters in an animal experiment: histopathology and SEM findings.

INTRODUCTION: Biocompatibility of implants in humans has been classified as "inert," "tolerated," and "bioactive." In shunt-treated patients, catheter-induced complications account for up to 70% of all hardware failures. Our objective was to study whether foreign body reactions to silicone shunt catheters in subcutaneous tissue and at their distal, intraperitoneal ends leading to occlusion can be reproduced in an animal model. MATERIALS AND METHODS: Twelve different silicone catheters were implanted in 6-week-old Wistar rats: (a) purely in the subcutaneous tissue and (b) through the subcutaneous tissue into the peritoneal cavity. One of the catheters was of our own design with a silicated surface. After 1 year, all catheters were explanted and were examined by histopathology and scanning electron microscopy (SEM). RESULTS: Histopathological analysis revealed the development of collagenous membranes and chronic immune reactions around the catheters. Completely organized intraluminal obliteration was seen in six intraperitoneally inserted catheters. SEM demonstrated calcifications and signs of biodegradation. Silicated catheters showed the most extensive calcifications. DISCUSSION: Hydrocephalus shunt catheters cannot be termed "inert" or "biotolerated." Rather, they must be regarded as "bio-active" implants. The extensive reaction on silicated catheters can act as reference to estimate the biocompatibility of surface modifications. The model proved appropriate for further studies.

Chalazion: racial risk factors for formation, recurrence, and surgical intervention.

OBJECTIVE: An association between race and formation of chalazion has yet to be objectively established. This study investigates race as a risk factor for chalazion and chalazion surgery. Understanding racial risk factors in formation of chalazion, recurrent chalazion, and chalazion requiring surgery (often with general anesthesia in children) informs decisions regarding eyelid hygiene, early topical medical therapy, and aggressiveness with oral antibiotic therapy for coexisting conditions such as blepharitis. METHODS: Demographic data was collected for all pediatric visits to the University of Wisconsin-Madison ophthalmology department from 2012-2019. Retrospective chart review was performed for the subset with chalazion. RESULTS: Of 28 433 minors, 584 had 1088 chalazia, a 2% overall rate. Chalazion was seen in 1.8% of non-Hispanic/Latino participants and 3.8% of Hispanic/Latino participants (p value <0.0001). Chalazion was seen in 1.7% of white participants, compared to 4.3% of American Indian or Alaska Native participants (p value <0.0001) and 4.0% of Asian participants (p value <0.0001). More than one chalazion was recorded in 31% of subjects without coexisting meibomian gland disease, blepharitis, or marginal keratitis, and in 56% (p < 0.0001) with one of these conditions. Repeated diagnoses of chalazion on separate encounters were seen in 17% without these conditions and in 33% (p < 0.0001) with one of these conditions. CONCLUSION: Hispanic/Latino, American Indian, and Asian participants developed chalazion at a rate higher than other racial/ethnic groups, whereas patients with meibomian gland disease or blepharitis are especially at risk for developing multiple chalazia on separate encounters. No group was more likely to require surgical intervention than any other.

Reliability of COVID-19 data: An evaluation and reflection.

IMPORTANCE: The rapid proliferation of COVID-19 has left governments scrambling, and several data aggregators are now assisting in the reporting of county cases and deaths. The different variables affecting reporting (e.g., time delays in reporting) necessitates a well-documented reliability study examining the data methods and discussion of possible causes of differences between aggregators. OBJECTIVE: To statistically evaluate the reliability of COVID-19 data across aggregators using case fatality rate (CFR) estimates and reliability statistics. DESIGN, SETTING, AND PARTICIPANTS: Cases and deaths were collected daily by volunteers via state and local health departments, as primary sources and newspaper reports, as secondary sources. In an effort to begin comparison for reliability statistical analysis, BroadStreet collected data from other COVID-19 aggregator sources, including USAFacts, Johns Hopkins University, New York Times, The COVID Tracking Project. MAIN OUTCOMES AND MEASURES: COVID-19 cases and death counts at the county and state levels. RESULTS: Lower levels of inter-rater agreement were observed across aggregators associated with the number of deaths, which manifested itself in state level Bayesian estimates of COVID-19 fatality rates. CONCLUSIONS AND RELEVANCE: A national, publicly available data set is needed for current and future disease outbreaks and improved reliability in reporting.

Evidence that the Ser192Tyr/Arg402Gln in cis Tyrosinase gene haplotype is a disease-causing allele in oculocutaneous albinism type 1B (OCA1B).

Oculocutaneous albinism type 1 (OCA1) is caused by pathogenic variants in the TYR (tyrosinase) gene which encodes the critical and rate-limiting enzyme in melanin synthesis. It is the most common OCA subtype found in Caucasians, accounting for ~50% of cases worldwide. The apparent 'missing heritability' in OCA is well described, with ~25-30% of clinically diagnosed individuals lacking two clearly pathogenic variants. Here we undertook empowered genetic studies in an extensive multigenerational Amish family, alongside a review of previously published literature, a retrospective analysis of in-house datasets, and tyrosinase activity studies. Together this provides irrefutable evidence of the pathogenicity of two common TYR variants, p.(Ser192Tyr) and p.(Arg402Gln) when inherited in cis alongside a pathogenic TYR variant in trans. We also show that homozygosity for the p.(Ser192Tyr)/p.(Arg402Gln) TYR haplotype results in a very mild, but fully penetrant, albinism phenotype. Together these data underscore the importance of including the TYR p.(Ser192Tyr)/p.(Arg402Gln) in cis haplotype as a pathogenic allele causative of OCA, which would likely increase molecular diagnoses in this missing heritability albinism cohort by 25-50%.

Early Holocene Scandinavian foragers on a journey to affluence: Mesolithic fish exploitation, seasonal abundance and storage investigated through strontium isotope ratios by laser ablation (LA-MC-ICP-MS).

At Norje Sunnansund, an Early Holocene settlement in southern Sweden, the world's earliest evidence of fermentation has been interpreted as a method of managing long-term and large-scale food surplus. While an advanced fishery is suggested by the number of recovered fish bones, until now it has not been possible to identify the origin of the fish, or whether and how their seasonal migration was exploited. We analysed strontium isotope ratios (87Sr/86Sr) in 16 cyprinid and 8 pike teeth, which were recovered at the site, both from within the fermentation pit and from different areas outside of it, by using laser ablation multi-collector inductively coupled plasma mass spectrometry. Our investigation indicates three different regions of origin for the fish at the site. We find that the most commonly fermented fish, cyprinids (roach), were caught in the autumn during their seasonal migration from the Baltic Sea to the sheltered stream and lake next to the site. This is in contrast to the cyprinids from other areas of the site, which were caught when migrating from nearby estuaries and the Baltic Sea coast during late spring. The pikes from the fermentation pit were caught in the autumn as by-catch to the mainly targeted roach while moving from the nearby Baltic Sea coast. Lastly, the pikes from outside the fermentation pit were likely caught as they migrated from nearby waters in sedimentary bedrock areas to the south of the site, to spawn in early spring. Combined, these data suggest an advanced fishery with the ability to combine optimal use of seasonal fish abundance at different times of the year. Our results offer insights into the practice of delayed-return consumption patterns, provide a more complete view of the storage system used, and increase our understanding of Early Holocene sedentism among northern hunter-fisher-gatherers. By applying advanced strontium isotope analyses to archaeological material integrated into an ecological setting, we present a methodology that can be used elsewhere to enhance our understanding of the otherwise elusive indications of storage practices and fish exploitation patterns among ancient foraging societies.

JUNB suppresses distant metastasis by influencing the initial metastatic stage.

The complex interactions between cells of the tumor microenvironment and cancer cells are considered a major determinant of cancer progression and metastasis. Yet, our understanding of the mechanisms of metastatic disease is not sufficient to successfully treat patients with advanced-stage cancer. JUNB is a member of the AP-1 transcription factor family shown to be frequently deregulated in human cancer and associated with invasion and metastasis. A strikingly high stromal JUNB expression in human breast cancer samples prompted us to functionally investigate the consequences of JUNB loss in cells of the tumor microenvironment on cancer progression and metastasis in mice. To adequately mimic the clinical situation, we applied a syngeneic spontaneous breast cancer metastasis model followed by primary tumor resection and identified stromal JUNB as a potent suppressor of distant metastasis. Comprehensive characterization of the JUNB-deficient tumor microenvironment revealed a strong influx of myeloid cells into primary breast tumors and lungs at early metastatic stage. In these infiltrating neutrophils, BV8 and MMP9, proteins promoting angiogenesis and tissue remodeling, were specifically upregulated in a JUNB-dependent manner. Taken together, we established stromal JUNB as a strong suppressor of distant metastasis. Consequently, therapeutic strategies targeting AP-1 should be carefully designed not to interfere with stromal JUNB expression as this may be detrimental for cancer patients.