Alteration of Notch signaling and functionality of adipose tissue derived mesenchymal stem cells in heart failure.

AIM: Circulating mesenchymal cells increase in heart failure (HF) patients and could be used therapeutically. Our aim was to investigate whether HF affects adipose tissue derived mesenchymal cell (adMSC) isolation, functional characteristics and Notch pathway. METHODS AND RESULTS: We compared 25 patients with different degrees of HF (11 NYHA classes I and II and 14 NYHA III and IV) with 10 age and gender matched controls. 100% adMSC cultures were obtained from controls, while only 72.7% and 35.7% from patients with mild or severe HF (p<0.0001). adMSC from HF patients showed higher markers of senescence (p16 positive cells: 14±2.3% in controls and 35.6±5.6% (p<0.05) and 69±14.7% (p<0.01) in mild or severe HF; γ-H2AX positive cells: 3.7±1.2%, 19.4±4.1% (p<0.05) and 23.7±3.4% (p<0.05) respectively), lower proliferation index (Ki67 positive cells: 21.5±4.9%, 13.2±2.8% and 13.7±3.2%, respectively), reduced pluripotency-associated genes (Oct4 positive cells: 86.7±4.9%, 55±12% (p<0.05) and 43.3±8.7% (p<0.05), respectively; NANOG positive cells: 89.8±3.7%, 39.6±14.4% (p<0.01) and 47±8.1%, respectively), and decreased differentiation markers (α-sarcomeric actin positive cells: 79.8±4.6%, 49±18.1% and 47±12.1% (p<0.05) and CD31-positive endothelial cells: 24.5±2.9%, 0.5±0.5% (p<0.001) and 2.3±2.3% (p<0.001), respectively). AdMSC from HF patients also showed reduced Notch transcriptional activity (lowered expression of Hey 1 and Hey 2 mRNAs). Stimulation with TNF-α of adMSC isolated from controls affected the transcription of several components of the Notch pathway (reduction of Notch 4 and Hes 1 mRNAs and increase of Notch 2 and Hey 1 mRNAs). CONCLUSIONS: In HF yield and functionality of adMSC are impaired and their Notch signaling is downregulated.

Evidence of LEF1 fetal-maternal interaction in cleft lip with or without cleft palate in a consistent Italian sample study.

Epithelial mesenchymal transformation is considered a cardinal process in orofacial development. Several molecular players appear to be involved in this delicate mechanism; the activation of LEF1 transcription factor by transforming growth factor beta 3 seems to be a key step for the correct flow of events. The failure of orofacial processes during embryonic development may provoke cleft lip and/or cleft palate malformations. The scope of the present investigation was to verify whether genetic variants at LEF1 could influence the risk of orofacial clefting. The approach was a family based association study involving a total of 512 Italian patients and their parents, 401 having cleft lip with or without cleft palate (CL/P) and 111 with cleft palate only (CPO). Haplotype association analysis provided moderate evidence of an association with clefting (p 0.01). A log-linear likelihood-based method was used to verify maternal and foetal-maternal association. An association between the maternal genotype and the occurrence of CL/P was observed at two polymorphic loci, at rs10022956 (P = 0.0049) and rs10025431 (P = 0.0065) respectively, while a foetal-maternal effect modulating the risk of clefting was found at locus rs10025431 (P = 0.0071). These data further corroborate the importance of the mother's genotype with regard to susceptibility to malformations and early-onset diseases.

No association between polymorphisms in cubilin, a gene of the homocysteine metabolism and the risk of non-syndromic cleft lip with or without cleft palate.

Epidemiological studies have correlated lower maternal periconceptional levels of plasma folate and cobalamin with increased risk of delivering offspring presenting congenital malformations such as cleft lip with or without cleft palate (CL/P) or neural tube defects. A number of genetic studies aimed at correlating these biochemical levels or the occurrence of malformations with specific genetic defects or polymorphisms have been successfully performed. The cubilin gene (CUBN) codes for a carrier that plays a crucial role in cobalamin cell internalization. CUBN polymorphisms were previously found to be associated with spina bifida occurrence. In this work, a family-based association study was performed to test CUBN involvement in CL/P. A sample of 391 CL/P triads was investigated with three single nucleotide polymorphisms mapping on the cubilin gene. Association tests indicated no significant association between CL/P and marker alleles or marker haplotypes. No evidence of maternal effect and imprinting were obtained. These data suggest that CUBN is not involved in CL/P onset in the investigated Italian population.

Corrective rhinoplasty: medical and legal aspects.

Preoperative planning of any surgical procedure, especially rhinoplasty, is crucial in order to achieve an optimal outcome which will satisfy both the surgeon and patient. Use of imaging and the possibility of virtually displaying the changes planned by the surgeon definitely lead to a better understanding between doctor and patient. Although these new technologies have led to undeniable advantages, they have also generated unprecedented medical and legal problems. The Authors discuss a number of reflections on the legal consequences of image digital processing, the problems related to the storage of such images, and the appropriate way to draft an informed consent.

Pregnancy and tram-flap breast reconstruction after mastectomy: a case report.

A 38-year-old patient had a right radical (Patey) mastectomy for an infiltrating ductal carcinoma followed by chemotherapy and, one year later, a TRAM flap breast reconstruction. She was given clear and exhaustive information about the possible consequences of pregnancy, but despite this she became pregnant four months after the reconstruction. The pregnancy was taken to term despite the appearance at four months of a slow-growing abdominal hernia. In the light of our experience and that of others, we recommend an interval of at least 12 months between breast reconstruction with a TRAM flap and pregnancy.

[Breast reconstruction]. / La ricostruzione mammaria.

The ideal goal of every post-mastectomy breast reconstruction is to achieve a breast that is as "identical" as possible to the contralateral one in shape, size, consistency, mobility and degree of naturalness. At the same time, however, it is essential to rely on the simplest and safest reconstruction technique as far as the patient is concerned. Mastectomy and restoration of the lost morphology should be performed in a single operation whenever the opportunity arises and there are two reasons for this. First of all, by working this way reconstruction can be performed on an area that is completely free of any scar tissue. Secondly, there are psychological considerations involved, since this makes it possible to avoid the trauma of letting the patient seeing herself, even for just an instant, without her breast. The numerous different techniques used for the morphological reconstruction of breast volume can be classified into two basic groups: reconstruction with insertion of a prosthesis and reconstruction without a prosthesis. Both of these techniques make it possible to personalize surgical choices based on the patient's unique characteristics, in order to achieve a complete symmetry in relation to the contralateral breast.

[The postoperative expansion of a myocutaneous flap of the large dorsal muscle in postmastectomy breast reconstruction]. / L'espansione postoperatoria del lembo miocutaneo di muscolo gran dorsale nella ricostruzione mammaria postmastectomia.

BACKGROUND: Reconstructive breast surgery using the myocutaneous flap of the latissimus dorsi muscle with simultaneous insertion of a prosthesis represents a very useful procedure from a clinical standpoint. METHODS: Twenty-one breast reconstructions were performed using this type of surgery at the Plastic Surgery Clinic of the University of Udine between December 1993 and December 1995. RESULTS: Good aesthetic results can be obtained due to the relatively moderate functional and scarring complications, as well as to the vitality of the transposed tissue. The percentage of capsular contractures observed frequently in the past was lowered through the use of technologically-advanced prosthetic materials with a texturized surface. CONCLUSIONS: In addition, since the reconstructed breast can be expanded postoperatively through definitive Becker expanders, aesthetically satisfactory results can be achieved even with contralateral ptosic and/or large breasts.

Myofasciocutaneous expansion in the tegumentary rehabilitation of the amputation stump.

In post-traumatic amputations, in particular in leg amputations, the presence of an unstable scar or of ulcerous areas that are difficult to heal in the site of the amputation stump constitutes a repair problem that is not easily solved. In this specific field of reconstruction skin expansion has earned significant agreement, and is even considered to be a safe and reliable method for the surgical rehabilitation of the amputation stump, allowing us to obtain high-quality tegumentary covering, with no excessive thickness, that adapts well to the prosthesis and to the underlying skeleton, and that is characterized by good sensitivity of protection. Tegumentary amplification may, in some cases, also become myocutaneous, allowing us to obtain more tissue thickness for protection in patients submitted to chronic loading that lasts longer and in those with thin teguments, in which traditional expansion could determine excessive narrowing of the tegumentary covering, following atrophy caused by compression of the subcutaneous adipose tissue.

Spiradenocarcinoma: morphological and immunohistochemical features.

Malignant transformation of spiradenoma is a very rare event and its biologic features have not been fully described. Clinically, these tumours are characterized by the rapid enlargement of a long-standing cutaneous nodule. Microscopic examination discloses the presence of the two components, benign and malignant, whose morphological distinction is not always easy. We report the occurrence of a spiradenocarcinoma within a pre-existing spiradenoma of the cheek in a 48 year old man and review the previously reported cases especially on the basis of their morphological and immunohistochemical characteristics.

[Anesthesiologic problems in patients with Launois-Bensaude-Madelung disease. Clinical case]. / Problematiche anestesiologiche in pazienti affetti da malattia di Launois-Bensaude-Madelung. Caso clinico.

Authors report a Launoise-Bensaude-Madelung disease case, in a 64 year old man, admitted to a Plastic Surgical Department for obesity, dysphonia, dysphagia, dyspnea. Early symptoms appeared 20 years before Hospital admission. Lipomatous tissue occupied nape, mandible, neck and shoulders. Surgical exeresis of lipomatous tissue under general anesthesia needed for the patient. Neck movements and mouth opening were short (Mallampati Score = 4); a neck computed tomography showed a tracheal compression and right displacement. Tracheal intubation was considered difficult or impossible. Nose-tracheal intubation was performed using a pediatric fiberoptic instrument as guide for a small gauge tracheal tube. Tracheal stenosis required many attempts for correct nose-tracheal intubation. Fiberoptic instrument as guide for tracheal tube can be useful for patients with Launoise-Bensaude-Madelung disease, when tracheal intubation is considered difficult or impossible. Knowledge of fiberoptic tracheal intubation techniques is mandatory for anesthesiologists, allowing tracheal intubation in patients with anatomical variations of mouth or upper respiratory airways.

[Scrotal reconstruction by cutaneous expansion]. / Reconstrucción del escroto mediante expansión cutánea.

We report on a patient who underwent scrotal reconstruction by cutaneous expansion, a new technique utilized to repair the male genital area. The tissue obtained with this technique permits creating a scrotal pouch and completely free movement of the testes. Unlike the other methods, sufficient skin can be obtained with the exparsion technique. The morphological result achieved with this technique is clearly superior. The absence of scars is underscored.

[A nasal reconstruction via the expansion of the tissue]. / La ricostruzione nasale mediante espansione cutanea.

The techniques that offer the best results in total nasal reconstruction are those that use frontal skin. Even if traditional procedures meet most of the reconstructive needs, they are often limited by the size of the tissues to be transplanted and by the scarring they leave in the donor area. The stiffness and scarce pliability of the frontal skin hinder fine modelling of the alae, of the columella and of the nasal tip. Expanded forehead flaps to day offer a substantial advantage. In the period from 1986 to 1991 total nasal reconstruction was performed in 12 patients (4 males and 8 females). Their ages ranged from 4 to 69 years. Four patients presented a recurrence of a basal cell carcinoma, 1 patient presented a trichoepithelioma. In 3 cases an arteriovenous vascular malformation was present, in 1 there were radiodermatitic lesions while in another 3 cases there was extensive scarring. In all cases a rectangular skin expander without base and with a remote filling port was used. The nominal volume ranged from 100 cc to 260 cc. All expanders were introduced through a vertical hair incision in the subgaleal plane, except in 1 case in which it was placed over the frontal muscle. The mean time of expansion was 79 day. The mean volume at the end of the expansion was 283 cc. The results obtained are presented and the advantages of the technique are exposed. These are: 1) the possibility of obtaining forehead flaps wide enough to cover large defects, 2) the direct closure of the donor area even if large flaps are raised, and 3) the improved pliability of the expanded frontal skin. A minor complication present in all the patients treated was moderate pain during inflation of the expander, easily controlled with common analgesics. A bony crest was frequently observed around the base of the skin expander which, however, was easily removed with a curette. Only in one case was a major complication observed; that of a preoperative infection necessitating removal of the expander, which was reapplied a few months later in order to complete the initial reconstructive program.

Skin expansion in head and neck reconstructive surgery.

We used a tissue-expansion technique to achieve soft-tissue reconstruction in congenital and acquired defects of head and neck. One hundred and fifty-five head and neck reconstructions have been performed, with a 6 percent rate of major complications. We present our planning and surgical approach for single areas. From a technical point of view, the only major difference from the rest of literature is the approach to the problem of periprosthetic capsule. In fact, we believe that complete removal of the capsule, which can be achieved without vascular risk thanks to a special technique, is necessary to obtain the best results.

Long-term histopathologic evaluation of human expanded skin.

Forty-two patients operated on by skin expansion have been contacted after a mean time of 25 months from the last surgery. Two biopsies have been taken from the expanded area of each patient. In 12 patients it has been possible to obtain a similar sampling from the opposite, nonexpanded area of the body. The samples underwent optic microscopy and cell kinetic and DNA content investigations. The epidermal structure of the followed-up skin, compared with the skin of the opposite side of the body, looks normal. The mitotic activity of the epidermal cells has returned to the values of preexpanded skin. The dermis shows a low degree of elastosis and zonal fragmentation of elastic fibers. The hypodermis, where the expander capsule was removed during the last surgery, does not show an accentuated fibrosis.

Cell kinetics and DNA content (ploidy) of human skin under expansion.

One question which still remains unanswered concerning expansion of human skin is whether expansion simply stretches the skin or whether it induces a real neo-production of epithelium. To evaluate the proliferative activity of the human skin before and after expansion, we performed an autoradiographic study and a densitometric analysis on sections of skin biopsy from 11 patients. The results of the cellular kinetic study showed that expansion induces an increase in the epidermal proliferative activity. On the contrary, the densitometric analysis did not show a parallel increase of the proliferating cells after expansion. This discrepancy is discussed in terms of the modified morphology of the tissue. The proliferative activity of skin in the back region increases more than in that of the neck. Preliminary results have shown that the proliferative activity increases with time of expansion in days. The presence of a low percentage of cells with altered DNA content indicates, at least over the short term, the absence of dysplastic processes in expanded skins.

The temporalis osteomuscular flap versus the free calvarial bone graft. An experimental study in the growing rabbit.

The authors have conducted an experimental study on the evolution of temporalis osteomuscular flaps compared with calvarial free bone grafts in young rabbits. The evaluation of the results has been performed with macroscopic, radiological and histological investigations in an early and late postoperative phase. This study has confirmed that the bone flaps remain viable and are characterized by a normal evolution. There is no evidence of anomaly of craniofacial growth nor asymmetry when compared with the contralateral side. On the other hand, the free bone grafts show the typical signs of necrosis and resorption. Some asymmetries are produced in comparison with the contralateral side, these being particularly evident in the two-months samples, while the overall craniofacial growth is not seriously impaired. This experimental study confirms the reliability of the temporalis osteomuscular flap in facial reconstruction.