Biochemische und molekularbiologische Charakterisierung von RAIP, einem neuen ER-lokalisierten proapoptotischen Protein

Charakterisierung eines zuvor funktionell unbeschriebenen Genes, das in einem genetischen Screen nach proapoptotischen Genen isoliert wurde und RAIP genannt wurde. Verifizierung der proapoptotischen Eigenschaften mit mehreren Apoptose-Assays in humanen Zelllinien, Nachweis der Lokalisation im ER in Kulturzellen, Eingrenzung eines 63 Aminosäure-Reste grossen proapoptotischen Fragmentes, Isolierung von drei Interaktionspartnern (Ferritin, SRp40, SIRTUIN 7)

Laivelio lūžio rekonstrukcija šonkaulio autotransplantatu: klinikinis atvejis ir literatūros apžvalga

Objective. To evaluate the results of scaphoid bone proximal pole reconstruction with rib osteochondral autograft due to comminuted scaphoid fracture. Material and methods. We present a clinical case of fragmented scaphoid bone proximal pole fracture reconstruction by rib osteochondral autograft. The modified wrist function score of Green and O’Brien and Quick Disabilities of the Arm, Shoulder and Hand (QuickDASH) outcome measuring scales were used for clinical evaluation before and 6 months after the reconstruction. Additio­nally, a literature review was conducted for case reports and previous literature reviews describing scaphoid bone proximal pole fracture surgical treatment. Medline (PubMed), ScienceDirect and UpToDate databases were used. Results. Conventional treatment methods for the treatment of comminuted proximal pole scaphoid bone fractures are often inappropriate due to technical issues or potential adverse outcomes. In these cases, reconstruction with rib autograft is possible. The study patient’s modified wrist function score of Green and O’Brien increased from 75 to 95 points out of 100 at 6 months postoperatively, and the Quick Disabilities of the Arm, Shoulder and Hand (QuickDASH) score decreased from 13.64 to 4.55 points. The results of this technique have been investigated in several studies (Sandow, 1998, 2001; Veitch et al., 2007). All subjects (22, 47 and 14 patients, respectively), except one, experienced improvement of wrist function – enhanced wrist movement, grip strength, reduced pain and restored wrist function to the pre-injury performance level. Conclusions. Scaphoid bone proximal pole fragmented fracture reconstruction with osteochondrial rib autograft achieves favorable recovery of wrist function and avoids complications or unfavorable functional consequences of alternative surgical procedures.Tikslas. Įvertinti laivelio proksimalinio poliaus rekonstrukcijos šonkaulio osteochondriniu autotransplantatu rezultatus, gydant skeveldrinį lūžį. Tyrimo metodai. Pateikiamas laivelio proksimalinio poliaus skeveldrinio lūžio rekonstrukcijos šonkaulio osteochondriniu autotransplantatu klinikinis atvejis. Klinikiniam paciento įvertinimui prieš operaciją ir praėjus 6 mėn. po atliktos rekonstrukcijos pasitelktas modifikuotas Greeno ir O’Brieno riešo funkcijos balas ir Quick Disabilities of the Arm, Shoulder and Hand (QuickDASH) vertinimo skalė. Taip pat, naudojantis Medline (PubMed), ScienceDirect ir UpToDate duomenų bazėmis, atlikta mokslinės literatūros apžvalga, apimanti laivelio proksimalinio poliaus lūžių chirurginio gydymo metodus ir galimybes. Rezultatai. Esant laivelio proksimalinio poliaus lūžiams, ypač skeveldrinio pobūdžio, tradiciniai gydymo metodai dėl techninių atlikimo principų ar numatomų nepalankių išeičių dažnai yra netinkami. Minėtais atvejais galima rekonstrukcija šonkaulio osteochondriniu autotransplantatu. Tirtam pacientui nustatytas modifikuotas Greeno ir O’Brieno riešo funkcijos balas, praėjus 6 mėn. po rekonstrukcinės operacijos, padidėjo nuo 75 iki 95 balų iš 100, o taikant Quick Disabilities of the Arm, Shoulder and Hand (QuickDASH) apskaičiuotas balas sumažėjo nuo 13,64 iki 4,55 balų. Minimų metodikų išeitys atskleidžiamos ir keliuose užsienio tyrimuose (Sandow, 1998, 2001; Veitch et al., 2007). Šių tyrimų duomenimis, atlikus laivelio rekonstrukciją osteochondriniu šonkaulio autotransplantatu, visų tiriamųjų (atitinkamai 22, 47 ir 14 pacientų), išskyrus vieną pacientą, judesio amplitudė ir sugriebimo jėga padidėjo, sumažėjo skausmas, tiriamieji atgavo prieš traumą buvusį darbingumą. Išvados. Atliekant laivelio proksimalinio poliaus skeveldrinio lūžio rekonstrukciją, kai kaulinio audinio defektas atkuriamas osteochondriniu šonkaulio autotransplantatu, pasiekiama palankių riešo funkcijos atkūrimo rezultatų, išvengiama alternatyvioms chirurginėms procedūroms būdingų komplikacijų ar nepalankių funkcinių padarinių

A new model for preclinical testing of dermal substitutes for human skin reconstruction

Background: Currently, acellular dermal substitutes used for skin reconstruction are usually covered with split-thickness skin grafts. The goal of this study was to develop an animal model in which such dermal substitutes can be tested under standardized conditions using a bioengineered dermo-epidermal skin graft for coverage. Methods: Bioengineered grafts consisting of collagen type I hydrogels with incorporated human fibroblasts and human keratinocytes seeded on these gels were produced. Two different dermal substitutes, namely Matriderm®, and an acellular collagen type I hydrogel, were applied onto full-thickness skin wounds created on the back of immuno-incompetent rats. As control, no dermal substitute was used. As coverage for the dermal substitutes either the bioengineered grafts were used, or, as controls, human split-thickness skin or neonatal rat epidermis were used. Grafts were excised 21days post-transplantation. Histology and immunofluorescence was performed to investigate survival, epidermis formation, and vascularization of the grafts. Results: The bioengineered grafts survived on all tested dermal substitutes. Epidermis formation and vascularization were comparable to the controls. Conclusion: We could successfully use human bioengineered grafts to test different dermal substitutes. This novel model can be used to investigate newly designed dermal substitutes in detail and in a standardized wa

Stenozuojančio tenosinovito operacinis gydymas: ankstyvieji ir vėlyvieji rezultatai, komplikacijos

Background. The thickening of A1 pulley of the tendon sheath limits the excursion of flexor tendon. Stenosing tenosynovitis causes finger movements dysfunction and pain. Objective. To analyze early and late outcomes of patients with stenosing tenosynovitis after surgical treatment – anulotomy. Methods. All patients had standard surgical procedure – open anulotomy of A1 pulley. Pain (verbal pain scale), hand and arm function (QuickDASH) and complications were recorded before surgery, after 1 week, 3 months and 6 months post surgery. Results. There were 45 patients, 29 (64%) female, 16 (36%) male. The highest pain score was recorded before surgery median 5 (IQR 5). The lowest pain score median 2 (IQR 2) was recorded after 6 months post surgery. The difference of the results after 1 week, 3 months and 6 months was statistically significant p < 0.001. The worst hand and arm function was before surgery and 1 week post surgery. Accordingly: medians 52 (IQR 33) and 52 (IQR 35). Full hand function recovery was noticed after 6 months post surgery median 0 (IQR 11). The difference is statistically significant p < 0.001. Conclusions. Surgical treatment, open anulotomy is one of the most effective methods for stenosing tenosynovitis. After this procedure pain and hand function improves greatly. However, for some patients it might cause discomfort of the hand because of the scar’s sensitivity and location.Įvadas. Sustorėjęs A1 skaidulinis žiedas, esantis ties delnakaulio galva, neleidžia sausgyslei laisvai judėti kanale. Pasireiškus šiam sutrikimui, pacientai, lenkdami ar tiesdami pirštą, jaučia vietinį skausmą ir strigimą. Tikslas. Įvertinti pacientų, sergančių stenozuojančiu tenosinovitu, ankstyvuosius ir vėlyvuosius rezultatus, pritaikius atvirąjį chirurginį gydymą, atliekant anulotomiją. Metodika. Tiriamiesiems atlikta standartinė operacija – atviras chirurginis plaštakos piršto skaidulinio A1 žiedo atvėrimas (anulotomija). Skausmas (verbalinė skausmo skalė), plaštakos ir rankos funkcijos (QuickDASH) bei komplikacijos vertinta po operacijos praėjus 1 sav., 3 mėn. ir 6 mėn. Rezultatai. Tyrime dalyvavo 45 pacientai, iš jų – 29 (64 %) moterys, 16 (36 %) vyrų. Didžiausią skausmą pacientai jautė prieš operaciją (mediana – 5; TKN 5), mažiausią – praėjus 6 mėn. po operacijos (mediana – 2; TKN 2). Skausmo rezultatų skirtumas prieš operaciją ir praėjus 1 sav., 3 mėn. bei 6 mėn. po operacijos – statistiškai reikšmingas (p < 0,001). Rankos ir plaštakos funkcijos blogiausiai vertintos prieš operaciją ir praėjus 1 sav. po operacijos (mediana atitinkamai: 52 (TKN 33) ir 52 (TKN 35). Plaštakos funkcija pacientams atkurta praėjus 6 mėn. po operacijos (mediana – 0; TKN 11). Skirtumas statistiškai reikšmingas (p < 0,001). Išvados. Atviroji anulotomija – viena iš efektyviausių stenozuojančio tenosinovito gydymo metodikų. Atlikus šią operaciją, gerėja plaštakos funkcija, mažėja skausmas. Vis dėlto pooperaciniu laikotarpiu nemažai daliai pacientų kyla tam tikrų nepatogumų ar komplikacijų, iš kurių dažniausiai minėtinas nepatogus randas. Neretai pacientus vargina rando jautrumas

Riešo kanalo tunelinio sindromo diagnostinių metodų palyginimas

Background. Carpal tunnel syndrome is the most common and widespread peripheral neuropathy in the world. The diagnostic testing methods for the carpal tunnel syndrome are based on anamnesis data, objective and instrumental inspection. Electroneuromyography is the main instrumental test when carpal tunnel syndrome is suspected. An ultrasound imaging may also be performed. Objective. To determine the effectiveness, sensitivity and specificity of instrumental diagnostics methods used to diagnose the carpal tunnel syndrome, and to compare them. Methods. Diagnostic testing – ultrasound and electroneuromyography was applied to persons under study. The area of the median nerve (mm2) was assessed during the ultrasound examination. The results of the electroneuromyography study evaluated the response speed of the sensory impulse (ms), the speed of the motor impulse response (ms). Later, the tests, their sensitivity and specificity were evaluated and compared. Results. 30 patients participated in the study, of which 26 (86.7%) were women, 4 (13.3%) were men. Electroneuromyography was found to be both sensitive and specific for a measure of sensory propagation velocity, 76.2 and 75%, respectively (p = 0.042); of motor propagation speed – 100 and 75% (p = 0.040). Ultrasonography is sensitive (87.5%) but nonspecific (66.7%) (p = 0.008). The study showed that the ultrasound dimension strongly, directly and reliably correlates with the sensory impulse response rate (p < 0.001), and the ultrasound dimension moderately, directly and reliably correlates with the motor impulse response rate (p < 0.001). Conclusions. Carpal tunnel syndrome can be suspected by ultrasound measurement of the area of the median nerve, but the test is only sensitive but non-specific. Electroneuromyography can confirm the diagnosis of carpal tunnel syndrome, as the test is both sensitive and specific. Comparing the studies, a strong, direct and reliable relationship between the results of the ultrasound and the electroneuromyography testing was established.vadas. Riešo kanalo sindromas – labiausiai paplitusi periferinių nervų neuropatija pasaulyje. Riešo kanalo tunelinio sindromo diagnostika paremta anamnezės duomenimis, objektyviu ištyrimu ir instrumentiniais tyrimo metodais. Pagrindinis instrumentinis tyrimas, įtariant riešo kanalo sindromą, yra elektroneuromiografija. Taip pat gali būti atliekamas ultragarsinis tyrimas. Tikslas – įvertinti pacientams, sergantiems riešo kanalo sindromu, taikytų instrumentinių diagnostinių metodų efektyvumą, jautrumą bei specifiškumą ir palyginti šiuos metodus tarpusavyje. Metodika. Tiriamiesiems atlikti diagnostiniai – ultragarsinis ir elektroneuromiografijos – tyrimai. Ultragarsinio tyrimo metu vertintas vidurinio nervo plotas (mm2). Elektroneuromiografijos tyrimu vertintas sensorinio impulso atsako greitis (ms) ir motorinio impulso atsako greitis (ms). Vėliau abu diagnostiniai tyrimai įvertinti atskirai, išskirtas jų jautrumas ir specifiškumas, metodai palyginti tarpusavyje. Rezultatai. Tyrime dalyvavo 30 pacientų, iš jų – 26 (86,7 %) moterys, 4 (13,3 %) vyrai. Nustatyta, kad elektroneuromiografijos tyrimas yra ir jautrus, ir specifiškas: sensorinio impulso atsako sklidimo greičio matmuo atitinkamai 76,2 % ir 75 % (p = 0,042); motorinio impulso atsako sklidimo greičio matmuo – 100 % ir 75 % (p = 0,040). Ultragarsinis tyrimas yra jautrus (87,5 %), tačiau nespecifiškas (66,7 %) (p = 0,008). Tyrimas parodė, kad ultragarsinis matmuo stipriai, tiesiogiai ir patikimai koreliuoja su sensoriniu impulso atsako greičiu (p < 0,001). Taip pat ultragarsinis matmuo vidutiniškai, tiesiogiai ir patikimai koreliuoja su motoriniu impulso atsako greičiu (p < 0,001). Išvados. Ultragarsu matuojant vidurinio nervo plotą, pacientui galima įtarti riešo kanalo sindromą, tačiau tyrimas yra tik jautrus, bet nespecifiškas. Atlikus elektroneuromiografijos tyrimą, riešo kanalo sindromo diagnozę galima patvirtinti, nes tyrimas yra ir jautrus, ir specifiškas. Tyrimus lyginant tarpusavyje, tarp ultragarsinio tyrimo ir elektroneuromiografijos tyrimo rezultatų nustatytas stiprus, tiesioginis ir patikimas ryšys

Alkūnės kanalo sindromo chirurginio gydymo rezultatai

Background. Cubital tunnel syndrome is the second most commonly diagnosed compressive neuropathy of the upper extremity after carpal tunnel syndrome and the most common neuropathy of the ulnar nerve. Objective. To evaluate the results of surgical treatment of patients with cubital tunnel syndrome by performing an open decompression in situ. Methods. All subjects underwent a standard operation – open surgical opening of the elbow canal (decompression in situ). On the day of surgery, after 3 months, after 6 months, pain (verbal pain scale), hand and arm function (QuickDASH), complications are evaluated. Results. There were 44 patients, 16 (36%) female, 28 (64%) male. In 50% of the patients, electroneuromyography revealed a severe degree of ulnar nerve damage, in the rest – a moderate degree of damage. Hand function for women 3 months after surgery improved by 42.046 points (according to QuickDASH), while in men it was 15.454 points (p < 0.05). Meanwhile, hand function for woman 3 months after surgery improved by 45.833 points, while in men it was 20.000 points (p < 0.05). Statistically significant differences between the groups with a moderate and severe degree of damage were found only when assessing pain according to the VAS scale at 3 months after surgery (mean VAS scores 1.2 and 2.8 respectively). A positive correlation was also observed between age and improvement in hand function between 3–6 months after surgery (p < 0.05). In older people, improvement in hand function is observed after a longer period of time after surgery, and a more pronounced improvement in hand function with a severe degree of damage (comparing function before surgery and 6 months after surgery) is experienced by younger people. Conclusions. In situ decompression of the ulnar nerve is one of the most effective methods of treating ulnar neuropathy. With this method an improvement in the function of the hand and a decrease in pain are observed.Įvadas. Alkūnės kanalo tunelinis sindromas – antra po riešo kanalo sindromo dažniausiai diagnozuojama kompresinė viršutinės galūnės neuropatija ir dažniausiai nustatomas alkūninio nervo neurologinis sutrikimas. Tikslas – įvertinti pacientų, operuotų dėl alkūnės kanalo sindromo, chirurginio gydymo rezultatus, atliekant atvirąją dekompresiją. Metodika. Tiriamiesiems atlikta standartinė operacija – atvirasis chirurginis alkūnės kanalo atvėrimas (dekompresija in situ). Operacijos dieną ir po operacijos praėjus 3 mėn. ir 6 mėn. vertintas skausmas (verbalinė skausmo skalė), plaštakos ir rankos funkcija (QuickDASH), komplikacijos. Rezultatai. Klinikiniame tyrime dalyvavo 44 pacientai, iš jų – 28 (64 %) vyrai ir 16 (36 %) moterų. 50 proc. pacientų elektroneuromiografijos tyrimu nustatytas sunkaus laipsnio alkūninio nervo pažeidimas, likusiems – vidutinio laipsnio pažeidimas. Praėjus 3 mėn. po operacijos, moterims plaštakos funkcija pagerėjo vidutiniškai 42,046 balo (QuickDASH), vyrams – 15,454 balo (p < 0,05). Praėjus 6 mėn. po operacijos, rankos funkcija, palyginti su rezultatu prieš operaciją, moterims pagerėjo vidutiniškai 45,833 balo, vyrams – 20,000 balo (p < 0,05). Statistiškai reikšmingas skirtumas tarp vidutinio ir sunkaus laipsnio pažeidimų grupių nustatytas tik vertinant skausmą, remiantis VAS skale, praėjus 3 mėn. po operacijos (VAS balų vidurkiai – atitinkamai 1,2 ir 2,8). Praėjus 3–6 mėn. po operacijos, rankos funkcijos pagerėjimas teigiamai koreliavo su tiriamųjų amžiumi (p < 0,05). Vyresniems pacientams rankos funkcijos pagerėjimas nustatytas praėjus daugiau laiko po operacijos. Didesnį rankos funkcijos pagerėjimą (lyginant funkciją prieš operaciją ir praėjus 6 mėn. po operacijos), esant sunkiam pažeidimo laipsniui, patiria jaunesni asmenys. Išvados. In situ alkūninio nervo dekompresija – viena iš efektyviausių alkūninio nervo neuropatijos gydymo metodikų. Taikant šį chirurginio gydymo metodą, pagerėja plaštakos funkcija, sumažėja skausmas

Skingineering I: engineering porcine dermo-epidermal skin analogues for autologous transplantation in a large animal model

Background: Extended full thickness skin defects still represent a considerable therapeutic challenge as ideal strategies for definitive autologous coverage are still not available. Tissue engineering of whole skin represents an equally attractive and ambitious novel approach. We have recently shown that laboratory-grown human skin analogues with near normal skin anatomy can be successfully transplanted on immuno-incompetent rats. The goal of the present study was to engineer autologous porcine skin grafts for transplantation in a large animal model (pig study=intended preclinical study). Materials and methods: Skin biopsies were taken from the pig's abdomen. Epidermal keratinocytes and dermal fibroblasts were isolated and then expanded on culture dishes. Subsequently, highly concentrated collagen hydrogels and collagen/fibrin hydrogels respectively, both containing dermal fibroblasts, were prepared. Fibroblast survival, proliferation, and morphology were monitored using fluorescent labelling and laser scanning confocal microscopy. Finally, keratinocytes were seeded onto this dermal construct and allowed to proliferate. The resulting in vitro generated porcine skin substitutes were analysed by H&E staining and immunofluorescence. Results: Dermal fibroblast proliferation and survival in pure collagen hydrogels was poor. Also, the cells were mainly round-shaped and they did not develop 3D-networks. In collagen/fibrin hydrogels, dermal fibroblast survival was significantly higher. The cells proliferated well, were spindle-shaped, and formed 3D-networks. When these latter dermal constructs were seeded with keratinocytes, a multilayered and partly stratified epidermis readily developed. Conclusion: This study provides compelling evidence that pig cell-derived skin analogues with near normal skin anatomy can be engineered in vitro. These tissue-engineered skin substitutes are needed to develop a large animal model to establish standardized autologous transplantation procedures for those studies that must be conducted before "skingineering” can eventually be clinically applie

Skingineering II: transplantation of large-scale laboratory-grown skin analogues in a new pig model

Background: Tissue engineering of skin with near-normal anatomy is an intriguing novel strategy to attack the still unsolved problem of how to ideally cover massive full-thickness skin defects. After successful production of large, pig cell-derived skin analogues, we now aim at developing an appropriate large animal model for transplantation studies. Materials and methods: In four adult Swiss pigs, full-thickness skin defects, measuring 7.5×7.5cm, were surgically created and then shielded against the surrounding skin by a new, self-designed silicone chamber. In two animals each, Integra dermal regeneration templates or cultured autologous skin analogues, respectively, were applied onto the wound bed. A sophisticated shock-absorbing dressing was applied for the ensuing 3weeks. Results were documented photographically and histologically. Results: All animals survived uneventfully. Integra healed in perfectly, while the dermo-epidermal skin analogues showed complete take of the dermal compartment but spots of missing epidermis. The chamber proved effective in precluding ("false positive”) healing from the wound edges and the special dressing efficiently kept the operation site intact and clean for the planned 3weeks. Conclusion: We present a novel and valid pig model permitting both transplantation of large autologous, laboratory-engineered skin analogues and also keeping the site of intervention undisturbed for at least three postoperative weeks. Hence, the model will be used for experiments testing whether such large skin analogues can restore near-normal skin, particularly in the long term. If so, clinical application can be envisione

Engineered neural tissue for peripheral nerve repair

A new combination of tissue engineering techniques provides a simple and effective method for building aligned cellular biomaterials. Self-alignment of Schwann cells within a tethered type-1 collagen matrix, followed by removal of interstitial fluid produces a stable tissue-like biomaterial that recreates the aligned cellular and extracellular matrix architecture associated with nerve grafts. Sheets of this engineered neural tissue supported and directed neuronal growth in a co-culture model, and initial in vivo tests showed that a device containing rods of rolled-up sheets could support neuronal growth during rat sciatic nerve repair (5 mm gap). Further testing of this device for repair of a critical-sized 15 mm gap showed that, at 8 weeks, engineered neural tissue had supported robust neuronal regeneration across the gap. This is, therefore, a useful new approach for generating anisotropic engineered tissues, and it can be used with Schwann cells to fabricate artificial neural tissue for peripheral nerve repair

Analysis of blood and lymph vascularization patterns in tissue-engineered human dermo-epidermal skin analogs of different pigmentation

PURPOSE: Bioengineered dermo-epidermal skin analogs containing melanocytes represent a promising approach to cover large skin defects including restoration of the patient's own skin color. So far, little is known about the development of blood and lymphatic vessels in pigmented skin analogs after transplantation. In this experimental study, we analyzed the advancement and differences of host blood and lymphatic vessel ingrowth into light- and dark-pigmented human tissue-engineered skin analogs in a rat model. METHODS: Keratinocytes, melanocytes, and fibroblasts from light- and dark-pigmented skin biopsies were isolated, cultured, and expanded. For each donor, melanocytes and keratinocytes were seeded in ratios of 1:1, 1:5, and 1:10 onto fibroblast-containing collagen gels. The skin analogs were subsequently transplanted onto full-thickness wounds of immuno-incompetent rats and quantitatively analyzed for vascular and lymphatic vessel density after 8 and 15 weeks. RESULTS: The skin analogs revealed a significant difference in vascularization patterns between light- and dark-pigmented constructs after 8 weeks, with a higher amount of blood vessels in light compared to dark skin. In contrast, no obvious difference could be detected within the light- and dark-pigmented group when varying melanocyte/keratinocyte ratios were used. However, after 15 weeks, the aforementioned difference in blood vessel density between light and dark constructs could no longer be detected. Regarding lymphatic vessels, light and dark analogs showed similar vessel density after 8 and 15 weeks, while there were generally less lymphatic than blood vessels. CONCLUSION: These data suggest that, at least during early skin maturation, keratinocytes, melanocytes, and fibroblasts from different skin color types used to construct pigmented dermo-epidermal skin analogs have distinct influences on the host tissue after transplantation. We speculate that different VEGF expression patterns might be involved in this disparate revascularization pattern observed