Enhancer landscape of lung neuroendocrine tumors reveals regulatory and developmental signatures with potential theranostic implications.

Well-differentiated low-grade lung neuroendocrine tumors (lung carcinoids or LNETs) are histopathologically classified as typical and atypical LNETs, but each subtype is still heterogeneous at both the molecular level and its clinical manifestation. Here, we report genome-wide profiles of primary LNETs' cis-regulatory elements by H3K27ac ChIP-seq with matching RNA-seq profiles. Analysis of these regulatory landscapes revealed three regulatory subtypes, independent of the typical/atypical classification. We identified unique differentiation signals that delineate each subtype. The "proneuronal" subtype emerges under the influence of ASCL1, SOX4, and TCF4 transcription factors, embodying a pronounced proneuronal signature. The "luminal-like" subtype is characterized by gain of acetylation at markers of luminal cells and GATA2 activation and loss of LRP5 and OTP. The "HNF+" subtype is characterized by a robust enhancer landscape driven by HNF1A, HNF4A, and FOXA3, with notable acetylation and expression of FGF signaling genes, especially FGFR3 and FGFR4, pivotal components of the FGF pathway. Our findings not only deepen the understanding of LNETs' regulatory and developmental diversity but also spotlight the HNF+ subtype's reliance on FGFR signaling. We demonstrate that targeting this pathway with FGF inhibitors curtails tumor growth both in vitro and in xenograft models, unveiling a potential vulnerability and paving the way for targeted therapies. Overall, our work provides an important resource for studying LNETs to reveal regulatory networks, differentiation signals, and therapeutically relevant dependencies.

Evaluation of Virtual Reality in the Reduction of Pain During Dressing Changes in Patients with Burn Wounds: A pilot study.

Burn injuries are among the most common life-threatening injuries for which medical attention is sought, and are accompanied by intense, severe pain, particularly during treatment. Burn therapy pain management with opioid and non-opioid analgesics is often insufficient when administered alone. Virtual reality (VR) interfaces provide an immersive experience that has demonstrable therapeutic benefits, including distraction from, and reduction of, pain. In this interventional pilot study, we assessed the correlation between VR and passive distraction and pain tolerance during burn wound treatment. This pilot study assessed patients undergoing burn wound dressing changes while receiving both pharmacological and VR intervention at [Removed for masked review]. Questionnaires and evaluation forms were subjectively completed by both patients and medical staff before and after treatment, and clinical metrics were recorded throughout the treatment. Forty-one patients ≥18 years old and 76.9% male that had primarily undergone ≥4 dressing changes before the study were included. Correlations were found between VR engagement during treatment and a decrease in subjective levels of nausea, anxiety, and pain sensation. Furthermore, high levels of VR engagement were correlated with high levels of VR enjoyment. These results suggest that highly engaging and enjoyable VR interfaces may reduce sensations of anxiety and pain in burn patients during dressing changes. Furthermore, these data suggest that VR technology may be applied as an adjunct therapy to pharmacological treatment in the standardization of burn wound care management. Further studies with control groups and larger sample populations are needed for better quantification of these benefits.

Effect of cyclosporine A - tempol topical gel for the treatment of alopecia and anti-inflammatory disorders.

Over the years, extensive research has been carried out to develop new chemical entities for hair loss treatment. Despite these efforts, the newly developed topical and oral treatments have not proven to be curative. Hair loss can result from underlying mechanisms, such as inflammation and apoptosis around hair follicles. We have developed a nanoemulsion based on Pemulen gel for topical application, tentatively addressing both mechanisms. The novel formulation contains two well-known molecules: Cyclosporin A (CsA), an immunosuppressant calcineurin inhibitor, and Tempol, a potent antioxidant. The in vitro permeation study on human skin revealed that the CsA-Tempol gel formulation effectively delivered CsA into the skin's inner target layer, the dermis. The effects of the CsA-Tempol gel on hair regrowth were further demonstrated in the in vivo well-established androgenetic model induced in female C57BL/6 mice. The beneficial outcome was statistically confirmed by quantitative analysis of hair regrowth, measured by color density. The results were further supported by histology analysis. Our findings revealed a topical synergy effect, resulting in lower therapeutic concentrations of both actives unlikely to cause systemic side effects. Overall, our research suggests that the CsA-Tempol gel is a highly promising platform for treating alopecia.

LGR5 expressing skin fibroblasts define a major cellular hub perturbed in scleroderma.

Systemic sclerosis (scleroderma, SSc) is an incurable autoimmune disease with high morbidity and mortality rates. Here, we conducted a population-scale single-cell genomic analysis of skin and blood samples of 56 healthy controls and 97 SSc patients at different stages of the disease. We found immune compartment dysfunction only in a specific subtype of diffuse SSc patients but global dysregulation of the stromal compartment, particularly in a previously undefined subset of LGR5+-scleroderma-associated fibroblasts (ScAFs). ScAFs are perturbed morphologically and molecularly in SSc patients. Single-cell multiome profiling of stromal cells revealed ScAF-specific markers, pathways, regulatory elements, and transcription factors underlining disease development. Systematic analysis of these molecular features with clinical metadata associates specific ScAF targets with disease pathogenesis and SSc clinical traits. Our high-resolution atlas of the sclerodermatous skin spectrum will enable a paradigm shift in the understanding of SSc disease and facilitate the development of biomarkers and therapeutic strategies.

Levodopa-responsive dystonia caused by biallelic PRKN exon inversion invisible to exome sequencing.

Biallelic pathogenic variants in PRKN (PARK2), encoding the E3 ubiquitin ligase parkin, lead to early-onset Parkinson's disease. Structural variants, including duplications or deletions, are common in PRKN due to their location within the fragile site FRA6E. These variants are readily detectable by copy number variation analysis. We studied four siblings with levodopa-responsive dystonia by exome sequencing followed by genome sequencing. Affected individuals developed juvenile levodopa-responsive dystonia with subsequent appearance of parkinsonism and motor fluctuations that improved by subthalamic stimulation. Exome sequencing and copy number variation analysis were not diagnostic, yet revealed a shared homozygous block including PRKN. Genome sequencing revealed an inversion within PRKN, with intronic breakpoints flanking exon 5. Breakpoint junction analysis implicated non-homologous end joining and possibly replicative mechanisms as the repair pathways involved. Analysis of cDNA indicated skipping of exon 5 (84 bp) that was replaced by 93 bp of retained intronic sequence, preserving the reading frame yet altering a significant number of residues. Balanced copy number inversions in PRKN are associated with a severe phenotype. Such structural variants, undetected by exome analysis and by copy number variation analysis, should be considered in the relevant clinical setting. These findings raise the possibility that PRKN structural variants are more common than currently estimated.

Topical nano-encapsulated cyclosporine formulation for atopic dermatitis treatment.

Systemic cyclosporine A (CsA) therapy shows efficacy in the treatment of recalcitrant severe atopic dermatitis (AD) but elicits severe side-effects. Thus, a topical formulation of CsA nanocapsules (NCs), able to potentially bypass these drawbacks, was developed. CsA-NCs were shown to enhance drug penetration into the various layers of porcine ear skin. Furthermore, the encapsulated CsA was biologically active, as shown in vitro on mouse splenocytes, reflected by inhibition of both cell proliferation and of interleukin (IL)-2 secretion. Ex-vivo efficacy was demonstrated on human skin organ culture by markedly reducing pro-inflammatory cytokines secretion. Finally, CsA-NCs topical formulation elicited improved efficacy in terms of better preservation of the skin barrier integrity, a decrease of the systemic pro-inflammation markers and reduced skin inflammation. The overall results suggest that this original topical platform may provide a novel therapeutic tool of clinical significance compared to the existing topical therapeutic drugs in AD.

Breast Implant Q Fever as a Source of In-Hospital Transmission.

Herein, we describe the first case of mammary implant infection caused by Coxiella burnetii, resulting in delayed diagnosis and treatment and an in-hospital cross-transmission of Q fever to medical personnel.

Decision-Making Styles and Levels of Involvement Concerning Breast Reconstructive Surgery: An Israeli Study.

PURPOSE/OBJECTIVES: To address decision-making styles among breast cancer survivors considering breast reconstruction.
. DESIGN: A primary analysis of a cross-sectional sample among survivors who chose to have breast reconstruction to examine correlations among patient age, decision-making style, and the level of involvement of decision making.
. SETTING: Hadassah Medical Center in Jerusalem, Israel.
. SAMPLE: 70 women who had undergone breast reconstruction surgery in the past five years.
. METHODS: Participants completed decision-making style and demographic questionnaires and an assessment of their level of involvement in the decision-making process. 
. MAIN RESEARCH VARIABLES: Level of involvement in decision making, decision-making model between provider and patient, and decision-making styles were examined.
. FINDINGS: No correlation was found between four main decision-making styles and patient age or the extent of patient decision-making involvement and age. A statistically significant correlation was found between the level of involvement in decision making and the decision-making style of the patient.
. CONCLUSIONS: Nurses should assess patient decision-making styles to ensure maximum patient involvement in the decision-making process based on personal desires regardless of age.
. IMPLICATIONS FOR NURSING: Nurses working in breast cancer care must address the decision-making process of patients diagnosed with breast cancer, including the choice to undergo breast reconstruction after mastectomy. Nurses should understand the complex factors that influence a woman's decision-making style to best help with the decision.

Different modes of herpes simplex virus type 1 spread in brain and skin tissues.

Herpes simplex virus type 1 (HSV-1) initially infects the skin and subsequently spreads to the nervous system. To investigate and compare HSV-1 mode of propagation in the two clinically relevant tissues, we have established ex vivo infection models, using native tissues of mouse and human skin, as well as mouse brain, maintained in organ cultures. HSV-1, which is naturally restricted to the human, infects and spreads in the mouse and human skin tissues in a similar fashion, thus validating the mouse model. The spread of HSV-1 in the skin was concentric to form typical plaques of limited size, predominantly of cytopathic cells. By contrast, HSV-1 spread in the brain tissue was directed along specific neuronal networks with no apparent cytopathic effect. Two additional differences were noted following infection of the skin and brain tissues. First, only a negligible amount of extracellular progeny virus was produced of the infected brain tissues, while substantial quantity of infectious progeny virus was released to the media of the infected skin. Second, antibodies against HSV-1, added following the infection, effectively restricted viral spread in the skin but have no effect on viral spread in the brain tissue. Taken together, these results reveal that HSV-1 spread within the brain tissue mostly by direct transfer from cell to cell, while in the skin the progeny extracellular virus predominates, thus facilitating the infection to new individuals.

Characterization of factors that determine lentiviral vector tropism in skin tissue using an ex vivo model.

BACKGROUND: Lentiviral tropism to a solid tissue may be determined by receptor availability, the differentiation state of cells and the three-dimensional architecture of the tissue. METHODS: Using skin organ cultures, lentiviral vector tropism was compared with that of keratinocytes in cell culture. Furthermore, the tropism of lentiviral vector to mouse and human tissues was compared ex vivo, in attempt to validate the mouse skin as an experimental system for human gene therapy of skin diseases. RESULTS: The results obtained indicated that although early progenitor keratinocytes (keratin 15+ and p63+), when grown in culture are permissive to lentiviral vector, they are resistant to transduction in their native 'niche' in the skin tissue. Transiently amplifying keratinocytes (keratin 14+) on the other hand, are permissive to lentiviral vector transduction, in cell culture and in the skin, after separation of the epidermis from the dermis layer. Keratinocytes (keratin 14+) in the hair follicle of human skin are resistant to lentiviral transduction, even after partial digestion of the extracellular matrix collagen. By contrast, collagenase pretreatment of mouse tissue facilitated transduction of keratinocytes within the hair follicle. Because lentivirus pseudotyped by two envelopes (amphotropic murine leukemia virus and vesicular stomatitis virus G glycoprotein) display the same tropism, we suggest that receptor availability is not the critical factor in the pattern of skin tissue transduction. CONCLUSIONS: Taken together, the results obtained in the present study indicate that lentiviral vector tropism in the three-dimensional skin tissue is distinct from the tropism to keratinocytes in culture and is dependent on a complex interplay of extracellular restrictions.

Tropism of lentiviral vectors in skin tissue.

The skin is an attractive tissue for gene therapy applications to treat genetic disorders and to express systemically delivered transgenes encoding therapeutic proteins. Understanding the tissue tropism of vectors is a prerequisite for the design of gene therapy trials. Using an ex vivo system of organ culture, we studied factors that determined viral tropism to the epidermal and dermal cells in human and mouse skin. We applied in these studies a lentiviral vector pseudotyped with two glycoproteins that use different cell receptors (vesicular stomatitis virus glycoprotein [VSV-G] and amphotropic murine leukemia virus envelope). The extent of infection with the amphotropic pseudotype was much higher than that of VSV-G, especially at low multiplicities of infection. In contrast, the tropism of these two pseudotypes in skin tissues was similar; at low multiplicities the infection was limited to areas near the basal layer of the epidermis, whereas at high multiplicities the infection extended to the dermal layer. To overcome physical barriers in the skin, the epidermal and dermal layers were separated and infected. Whereas the human epidermis was readily infected, we could not detect infection of stem and early progenitor cells in their niche. In contrast, mouse epidermis was completely resistant to infection. Dermal cells of both species were readily infected with the two pseudotypes. Molecular analysis indicated that infection of mouse epidermal cells was restricted after proviral DNA synthesis and before integration. In conclusion, we show that lentiviral tropism in a solid tissue is dependent on several factors, extra- and intracellular, distinct of the cellular receptors.

Suicide bombing injuries: the Jerusalem experience of exceptional tissue damage posing a new challenge for the reconstructive surgeon.

BACKGROUND: Suicide bomb injuries vary in form and magnitude. From the onset of the second Palestinian "intifada" in October of 2000 until January of 2004, 577 victims of suicide bombings were admitted to the Hadassah-Hebrew University Medical Center. A single bomber carrying a handbag or belt containing multiple metal objects and explosives carried out most of the attacks. As a result, many of the victims suffered massive tissue destruction in addition to conventional blast injuries. METHODS: This article describes the management of this trauma-related "syndrome" of combined primary and high-magnitude secondary blast injury. RESULTS: The management of the extensive soft-tissue damage is described and two representative cases presented. CONCLUSION: Suicide bombing-related injuries in their present form are a true challenge for the reconstructive surgeon.

[Vacuum--a novel method for treating chronic wounds].

Chronic wounds represent a major health burden and drain on resources. In spite of advances in our understanding of chronic wound biology and the development of new treatments, there is no one form of treatment suitable for all wounds and patients. Many factors can impair healing--local and systemic. The current treatments include different ways of debridement, occlusive hydrocolloid dressings, topical growth factors, hyperbaric oxygen, bioengineered skin equivalents and more. In the last decade there is an increment in the use of negative pressure dressing (VAC--vacuum assisted closure system) in the treatment of chronic wounds from different etiologies--diabetic, venous and pressure sores. Our experience demonstrated good results in the treatment of diabetic and venous insufficiency ulcers, and disappointing results in the treatment of pressure sores. Promising results were demonstrated in the treatment of acute injuries, lowering the need for microvascular reconstruction. Prospective researches examining the efficacy and cost effectiveness of this treatment in comparison to other methods are crucial for wise and extensive use of the VAC system.