Diagnosis, genetic characterization and clinical follow up of mitochondrial fatty acid oxidation disorders in the new era of expanded newborn screening: A single centre experience.

INTRODUCTION: Mitochondrial fatty acid oxidation disorders (FAODs) are a heterogeneous group of hereditary autosomal recessive diseases included in newborn screening (NBS) program in Italy. The aim of this study was to analyse FAODs cases, identified either clinically or by NBS,for clinical and genetic characterization and to evaluate a five years' experience of NBS, in the attempt to figure out the complexity of genotype-phenotype correlation and to confirm the clinical impact of NBS in our centre experience. MATERIALS AND METHODS: We analysed FAODs patients diagnosed either by NBS or clinically, followed since February 2014 to April 2019 at the Regional Screening Centre and Inherited Metabolic Diseases Unit of Verona. Diagnosis was confirmed by plasma acylcarnitines, urinary organic acids, enzymatic and genetic testing. For not clear genotypes due to the presence of variants of uncertain significance, in silico predictive tools have been used as well as enzymatic activity assays. Patients underwent clinical, nutritional and biochemical follow up. RESULTS: We diagnosed 30 patients with FAODs. 20 by NBS: 3 CUD, 6 SCADD, 5 MCADD, 4 VLCADD, 2 MADD. Overall incidence of FAODs diagnosed by NBS was 1:4316 newborns. No one reported complications during the follow up period. 10 patients were diagnosed clinically: 2 CUD, 2 CPT2D, 1 VLCADD, 5 MADD. Mean age at diagnosis was 29.3 years. Within this group, complications or symptoms were reported at diagnosis, but not during follow-up. 12 mutations not previously reported in literature were found, all predicted as pathogenic or likely pathogenic. DISCUSSION AND CONCLUSIONS: Our study highlighted the great phenotypic variability and molecular heterogeneity of FAODs and confirmed the importance of a tailored follow up and treatment. Despite the short duration of follow up, early identification by NBS prevented diseases related complications and resulted in normal growth and psycho-motor development as well.

Abdominal Wall Reconstruction after Flap Surgery and the Effect on the Immune System.

BACKGROUND: The aim of our study was to investigate the impact of abdominal wall reconstruction surgery on tissue anatomy and to explore how flap surgery influences the patient's immune status. METHODS: Experimental abdominal wall defects were created in 8 Sus scrofa (swine) animal models. The animals were divided into two groups: 4 swine were euthanized one month after surgery for the biopsies retrieval purpose and the other 4 swine were kept alive and the collection of blood samples has been done 6 months after surgery. In order to evaluate the relative gene expression in operated-on animal cohorts we compared them with samples from 4 healthy swine used as controls. RESULTS: The inflammatory process was present in all types of repairs. Collagen I deposition was higher in the flap repairs. The expression level for the genes related to immune response after 6 months from surgery was relatively similar to the control group except minor alteration registered in the case of two swine models. CONCLUSION: Our findings indicate a less pronounced proinflammatory response to surgical trauma in animal models after flap surgery. The postoperative levels of the inflammatory cytokines did not show significant differences after abdominal wall reconstruction using flap surgery.

Prostate cancer between prognosis and adequate/proper therapy.

Knowing the indolent, non-invasive nature of most types of prostate cancer, as well as the simple fact that the disease seems more likely to be associated with age rather than with other factors (50% of men at the age of 50 and 80% at the age of 80 have it [1], with or without presenting any symptom), the big challenge of this clinical entity was to determine severity indicators (so far insufficient) to guide the physician towards an adequate attitude in the clinical setting. The risk of over-diagnosing and over-treating many prostate cancer cases (indicated by all the major European and American studies) is real and poses many question marks. The present paper was meant to deliver new research data and to reset the clinical approach in prostate cancer cases.

Immunotherapy and gene therapy in prostate cancer treatment.

There are few methods bringing several relatively recent advances in therapy of certain types of prostate cancer. Belonging to personalized therapies, they use cells (normal or pathologic) from the patient, modify and reintroduce them in the patient's body, leading to an increased efficiency against the neoplastic tissue, proving to increase the patient's lifespan and/ or tumor progression.

Vitamin B12 Administration by Subcutaneous Catheter Device in a Cobalamin A (cblA) Patient.

Cobalamin A deficiency (cblA) is an inherited disorder of intracellular cobalamin metabolism, caused by impaired 5'-deoxy-adenosylcobalamin (AdoCbl) synthesis. Hydroxocobalamin (OHCbl) is the cornerstone of cblA treatment because vitamin B12 may completely restore AdoCbl deficiency. Parenteral administration, intravenous, subcutaneous or intramuscular, is generally required to achieve effect. Daily injections represent a problem for the parents and the caregivers, and this may lead to poor compliance and scarce adherence to the long-term treatment.Our report describes the case of a patient with cblA deficiency, diagnosed by newborn screening, positively treated with daily OHCbl administration by a subcutaneous injection port (i-port advanceTM). After the insertion of the device, we checked methylmalonic acid (MMA) levels weekly for the first month and then monthly. MMA level remained always in the normal range.To date, placement of a subcutaneous catheter to minimize the pain related to parenteral vitamin B12 punctures has been described only in a patient with deficiency of the enzyme methylmalonyl-CoA mutase (MUT). No other experiences are described in the literature.Our case shows that OHCbl administration using a subcutaneous catheter is safe and effective even in patients with cblA deficiency. The use of subcutaneous devices may reduce difficulties in providing parenteral daily injections which is the main reason discouraging physicians and families to use such an invasive treatment. Moreover, our experience may be translated to other inherited metabolic disorders, such as cobalamin C (cblC) disease, which may require daily parenteral drug administration.

The etiopathogenesis of uterine fibromatosis.

Uterine fibroids or uterine leiomyomas are the most common benign tumors of the uterus among women of fertile age, while the etiology is still incompletely elucidated. The occurrence and development of the fibromatosis may be related to certain risk factors and genic mechanisms, although the exact causes are not yet fully known. The development of uterine fibroids is correlated not only with the metabolism and with the level of female sexual hormones, estrogen, and progesterone, but also with the number of these hormone receptors expressed on the surface of the myometrium. Proliferative effects of estrogen and progesterone may be exercised through proinflammatory factors (TNF alpha), growth factors (IGF1, IGF2, TGFbeta3 and betaFGF) or inhibitors of apoptosis (p53 suppression). A number of predisposing factors such as ethnicity - black skin, early menarche, nulliparity, caffeine and alcohol, chronic inflammation, obesity, were also identified. Approximately 40% of the uterine fibroids are caused by the same cytogenetic alterations found in the other tumor types such as kidney, lung, or leiomyosarcoma. As part of a system dysfunction, uterine fibromatosis was connected to other disorders such as AHT (arterial hypertension), endometrium adenocarcinoma, adenomyosis, endometriosis, diabetes mellitus, breast tumors, seemingly with a common causality. The action and effect of some hormonal imbalances over the various organs depend on the histological and local expression particularities of the various receptors, being the cause for many disorders, among which the uterine fibromatosis, coexisting or accompanying the later. This article examines and summarizes the latest data refreshed literature etiopathogenesis offering indicators of uterine fibroids.

Focal adenomyosis (intramural endometriotic cyst) in a very young patient - differential diagnosis with uterine fibromatosis.

Introduction. Adenomyosis is a widespread disease usually affecting the late reproductive years of the women's life, which has a great impact on their fertility. The most common form is diffuse adenomyosis, while focal adenomyosis, a cystic variant, is very rare, particularly in patients younger than 30 years old. Materials and methods. We reported a rare case of a 20-year-old Caucasian woman with cystic adenomyosis who was admitted in our service with severe chronic pelvic pain, dysmenorrhea, and menorrhagia, who had received conservative surgical treatment to preserve fertility and improve her obstetrical prognosis. Results and Discussions. Although the necrobiosis of a uterine fibroid was suspected preoperatively, the extemporaneous histopathological exam revealed adenomyosis associated with fibroleiomyoma with hyaline dystrophy and multiple foci of endometriosis of cystic formation in the wall of a young woman without any risk factors. Conclusion. Although a rare lesion in young patients, cystic adenomyosis should be considered when chronic pelvic pain is exacerbated during menstruation and is associated with a uterine tumor. In young patients, the differential diagnosis should be made with uterine malformations (hematometra), necrobiosis of uterine fibroids, pelvic endometriosis. The surgical treatment should be conservative with the excision of the lesion, always taking into account fertility preservation in young patients.

Curcumin delivered through bovine serum albumin/polysaccharides multilayered microcapsules.

The aim of the paper is to obtain and characterize k-carrageenan-chitosan dual hydrogel multilayers shell BSA gel microcapsules, as a carrier for curcumin, and as a possible antitumoral agent in biological studies. We used the CaCO3 template to synthesize non-toxic CaCO3/BSA particles as microtemplates by coprecipitating a CaCl2 solution that contains dissolved BSA, with an equimolar Na2CO3 solution. The microcapsules shell is assembled through a layer-by-layer deposition technique of calcium cross-linked k-carrageenan hydrogel alternating with polyelectrolite complex hydrogel formed via electrostatic interactions between k-carrageenan and chitosan. After the removal of CaCO3 through Ca(2+) complexation with EDTA, and by a slightly treatment with HCl diluted solution, the BSA core is turned into a BSA gel through a thermal treatment. The BSA gel microcapsules were then loaded with curcumin, through a diffusion process from curcumin ethanolic solution. All the synthesized particles and microcapsules were stucturally characterized by: Fourier Transform Infrared Spectroscopy, UV-Vis Spectrometry, X-ray diffraction, thermal analysis, fluorescence spectroscopy, fluorescence optical microscopy, confocal laser scanning microscopy and scanning electron microscopy. The behavior of curcumin loaded microcapsules in media of different pH (SGF, SIF and PBS) was studied in order to reveal the kinetics and the release profile of curcumin. The in vitro evaluation of the antitumoral activity of encapsulated curcumin microcapsules on HeLa cell line and the primary culture of mesenchymal stem cells is the main reason of the microcapsules synthesis as BSA-based vehicle meant to enhance the biodisponibility of curcumin, whose anti-tumor, anti-oxidant and anti-inflammatory properties are well known.

ORBITAL PSEUDOTUMOR CAN MIMIC GRAVES' OPHTHALMOPATHY.

CONTEXT: Orbital pseudotumor (OP) is a benign inflammatory process of the orbit with a large polymorphous lymphoid infiltrate, associated with fibrosis in variable amounts, localized or diffuse. Graves' ophthalmopathy (GO) is the most common cause of proptosis, unilateral or bilateral. CASE REPORT: We report a patient with unilateral proptosis who was initially treated with antithyroid drugs for an euthyroid Graves disease, but the extension of the investigation infirmed this. The MRI findings (inflammation of fat, muscle and the left lacrimal gland) in conjunction with the biopsy infirmed the initial diagnostic and confirmed the OP. CONCLUSIONS: Orbital pseudotumor is a rare disorder that can image and clinically mimics some inflammatory disease especialy Graves' ophthalmopathy. Orbital MRI represents the most important test for diagnostic. A negative TRAb in euthyroid cases of proptosis can be a serious starting point for investigating an OP.

ASSOCIATION BETWEEN LOW BONE MINERAL DENSITY, METABOLIC SYNDROME AND SEX STEROIDS DEFICIENCY IN MEN.

OBJECTIVE: To analyze the association between low bone mineral density (BMD), metabolic syndrome (MS) and sex hormones deficiency in men. METHODS: We included in this retrospective study 199 men with osteoporosis or osteopenia and 167 men with normal BMD as controls, aged between 55-85 years old. Patients' evaluation included: medical history and physical examination, X-ray of thoracic and lumbar spine, measuring BMD at hip and lumbar spine, serum glucose and lipid profile, serum levels of total testosterone (tT), free testosterone (fT) and estradiol (E2). RESULTS: The results revealed a significant association between low BMD and MS (p=0.011). Vertebral fractures were more frequently associated with MS (p=0.041). Patients with MS had lower vertebral BMD (p=0.037) and lower E2 levels (p=0.024) compared with those without MS. In men with MS, E2 deficiency can predict the value of vertebral and hip BMD. fT deficiency can predict only the value of hip BMD. CONCLUSIONS: A significant association between MS, low BMD, vertebral fractures and sex steroids deficiency, in particular E2 and fT was found. The presence of MS and sex hormones deficit can predict the reduction of BMD.

Measurement of Quality of Life after Total Parathyroidectomy in Patients with Secondary Hyperparathyroidism and End Stage Renal Disease.

BACKGROUND: Surgery is the definitive treatment of secondary hyperparathyroidism (2HPT) and end stage renal disease patients. The aim of this study to assess the impact of surgery on the evolution of quality of life (QoL) in patients with 2HPT, and to identify the variables that influence this evolution. METHODS: A series of 85 consecutive unselected patients underwent total parathyroidectomy for 2HPT in our clinic. QoL was measured using the Short-Form Health Survey(SF-36) and alleviation of symptoms was documented using an outcome tool (PAS score), based on visual analog scales, preoperatively, postoperatively and at 6 months. RESULTS: Preoperatively, patients had lower SF-36 scores than the general population in all 8 individual and 2 component summary scales, with significant decrease in the physical health scales(p 0.0001). Patients improved in all ten scales at 6 months follow-up, most significant being: Body Pain (45.02‚+-5.52 vs 33.12‚+-8.82, p 0.0001), Role-physical (41.00‚+-7.43 vs 33.46‚+-8.54, p 0.0001), Physical functioning (40.06 ‚+-7.77 vs 33.36 ‚+-10.84, p 0.0001). PAS Scores decreased from preoperatory levels of 569.99 ‚+- 136.45 to 372.20 ‚+- 104.62 at 7 days after surgery and furthermore at 292.64 ‚+- 85.16 at 6 months follow-up (p 0.0001). CONCLUSIONS: We found no correlation between preoparatory PTH or Calcium levels and clinical symptoms. Parathyroidectomy clearly alleviates symptomatology and improves QoL in 2HPT patients, with durable effect at 6 months.

Surgical Attitude in Patients with Secondary Hyperparathyroidism Undergoing Dialysis.

BACKGROUND: Secondary hyperparathyroidism (SHPT), develops, more or less in all the patients with chronic kidney disease. The pathology is even more severe as it intervenes in a suffering patient in whom the chronic kidney disease frequently associates severe comorbidities. General mortality is higher than in general population. The failure of the medical therapy is an indication for parathyroidectomy. METHODS: The study analyzed 200 patients with SHPT and chronic kidney disease, admitted in the clinic from October 2011 until January 2015.In this period, 179 (89.5 %) total-parathyroidectomies have been performed a long with 14 (7%) subtotal parathyroidectomies. Also 7 (3.5%) surgical interventions were incomplete. RESULTS: Overall mortality was 1% (2 patients) and postoperative specific morbidity 3.5% -4 local hemorrhagic complications and 3 cases of dysphonia have been encountered (12% if we include the reinterventions for recurrent hyperparathyroidism - 17 patients). CONCLUSIONS: Total parathyroidectomy is encumbered by a reduced number of postoperative complications and the risk of recurrent disease in almost nonexistent. The disadvantages of this surgical approach are the tendency of immediate postoperative hypocalcemia and long therm substitution with calcium and vitamine D.

Technical Factors Involved in Parathyroid Surgery.

We aimed to investigate the frequency of ectopic and supernumerary parathyroid glands in our series of renal hyperparathyroidism. From October 2011 to November 2014, 202 patients with chronic renal failure and advanced SHPT nonresponsive to medical therapy were hospitalized in the General Surgery Department of the Carol Davila Nephrology Hospital. These patients underwent a number of 188 (93%) total parathyroidectomies and a number of 14 patients (7%) subtotal parathyroidectomies. Of these 202 patients, reoperation was carried out for 14 patients (7%) in which we identified ectopic and supernumerary parathyroid glands. Operative details and pathology results were prospectively collected and reviewed after we obtained informed consent for data and pictures use. In 188 patients (93% cases), four or more parathyroid glands were removed at the first operation. In 14 cases (7%) high PTH level persisted after the initial operation. In 22 of them (11%), supernumerary glands were found at the first operation and in 6 of them (3%) at the second operation. We conclude that extensive cervical exploration in addition with preoperative imaging tests, parathyroid ultrasound; scintigraphy with Tc will reduce secondary hyperparathyroidism surgery.

The Role of Collagen Metabolism in the Formation and Relapse of Incisional Hernia.

Despite the continuous development of synthetic prosthetic meshes and their wide use, recurrent incisional hernias still appear in 5 to 20% of cases, with a linear incidence curve over the years, suggesting a multifactorial process rather than a simple failing technical repair as the underlying cause. Recent molecular biological research provide increasing evidence of connective tissue alterations such as a defective wound healing with impaired scarring process in patients with incisional hernia. Although there are some promising results, at present, in-depth understanding of the pathophysiological mechanisms and of the role that collagens play in the development and recurrence of incisional hernia is rather scarce. The aim of this systematic review is to summarize and evaluate the biochemical mechanisms involved in incisional hernia formation and recurrence, with a primary focus on collagen I to III ratio. Also, the consequences for surgical practice are discussed.

Spontaneous rupture of a splenic hydatid cyst with anaphylaxis in a patient with multi-organ hydatid disease.

Hidatid cysts of the spleen are a rare occurrence, the spleen being the third most common organ for the development of Echinococcus Granulosus. Splenic hydatid cysts are commonly part of multi-organ hydatid disease. Diagnosis is often established when investigating a splenomegaly or by chance during an unrelated consult. It can also be diagnosed after rupture, be it following trauma (the most common occurrence)or spontaneous. Splenic hydatid cyst rupture requires immediate action and is a life-threatening condition. It results, most often, in splenectomy. We present the case of a patient with multi-organ hydatid disease that presented with a ruptured splenic cyst and developed anaphylaxis. The case was resolved by splenectomy and recovered well.

Translating clinical research of Molecular Biology into a personalized, multidisciplinary approach of colorectal cancer patients.

Although multimodal treatment has brought important benefit, there is still great heterogeneity regarding the indication and response to chemotherapy in Stage II and III, and individual variations related to both overall survival and toxicity of new therapies in metastatic disease or tumor relapse. Recent research in molecular biology led to the development of a large scale of genetic biomarkers, but their clinical use is not concordant with the high expectations. The Aim of this review is to identify and discuss the molecular markers with proven clinical applicability as prognostic and/or predictive factors in CRC and also to establish a feasible algorithm of molecular testing, as routine practice, in the personalized, multidisciplinary approach of colorectal cancer patients in our country. Despite the revolution that occurred in the field of molecular marker research, only Serum CEA, Immunohistochemical analysis of mismatch repair proteins and PCR testing for KRAS and BRAF mutations have confirmed their clinical utility in the management of colorectal cancer. Their implementation in the current practice should partially resolve some of the controversies related to this heterogenic pathology, in matters of prognosis in different TNM stages, stage II patient risk stratification, diagnosis of hereditary CRC and likelihood of benefit from anti EGFR therapy in metastatic disease. The proposed algorithms of molecular testing are very useful but still imperfect and require further validation and constant optimization.

Stump appendicitis - an overlooked clinical entity.

Stump appendicitis is a rare disease, often overlooked. Thepossibility of stump appendicitis is not clear to all clinicians.It can represent a diagnostic dilemma if the treating physicianis not aware of this uncommon disease. Presenting symptomsare indistinguishable from those of primary appendicitis. Wepresent two cases of stump appendicitis operated in ourdepartment and their treatment. A heightened awareness ofthe possibility of this disease can lead to good initial surgical technique and prevention, and to early diagnosis in the eventof stump appendicitis.

Zenker's diverticulum, a rare cause of upper gastrointestinal bleeding.

UNLABELLED: The most common complication of Zenker's diverticulum is aspiration pneumonia, compression of the trachea and esophageal obstruction with large diverticulum, and increased risk of development of carcinoma. Thus bleeding occurs rarely, can be massive and life threatening, with ulceration being the most common cause. MATERIAL AND METHODS: We describe a patient with sever upper gastrointestinal bleeding as a result of a Zenker's diverticulum. A 75 year-old woman was referred to the emergency room and hospitalized for hematemesis, melena, asthenia and total dysphagia. In this particular case we preferred open technique because of the diverticulum dimensions and bleeding episode. Left cervicotomy was practiced on the anterior edge of the sternocleidomastoid muscle, being known that Zenker diverticulum extend into the left neck 90% of the time, fact also confirmed by radiology in this case. RESULTS: Postoperatively, the patient showed a complication free recovery. Five days after treatment the patient resumed nourishment. Several days later our patient was able to return home. Follow-up at 12 months after the operation showed complete recovery. Ulcer of the basis of Zenker's diverticulum is a rare entity and, only a few cases were reported in the literature to date. Omitting thecricomyotomy predisposes to fistula or diverticulum recurrence due to the persistence of a high pharyngeal intraluminal pressure that acts on the posterior wall just proximal to the upper esophageal sphincter. CONCLUSIONS: Zenker's diverticulum is an unusual site of origin for clinically significant upper gastrointestinal hemorrhage and differential diagnosis must include other more frequent causes of upper gastrointestinal bleeding. In our opinion, classicalsurgical therapy is indicated when distal esophageal imaging cannot be obtained during endoscopic examination, there is a large diverticulum or in an emergency setting when fast control over the bleeding source is required.

Necrotizing enterocolitis, a rare but severe condition with insidious postoperative complications.

UNLABELLED: Necrotizing enterocolitis (NEC) is one of the most frequent causes of gastrointestinal perforation in premature neonates, only few case series and reports being described in adult patients. Early in the course of the disease, superficial mucosal ulceration, sub mucosal edema and hemorrhage occur. Further progression leads to transmural necrosis leading sometimes to bowel perforation. Six cases encountered in our clinic in recent years led us to resume discussions on necrotizing enteritis, not because it is a rare disease, but due to the severe postoperative complications. MATERIAL AND METHODS: Our lot consisted of four stage 1 patients and two with Bell stage III NEC and severe intestinal injury, necrosis, and perforation. All of the patients were diagnosed preoperatory with other surgical conditions, like appendicitis with peritonitis, perforated duodenal ulcer or acute cholecystitis. RESULTS: We present to review two cases. For patients undergoing laparotomy, resection of the involved intestine mandates either enterostomy formation or primary anastomosis. An intermediate option is laparotomy with intestinal resection and delayed anastomosis 48 to 72 hours later. Because of the small number of patients in our lot, we cannot advise a certain surgical treatment, but a strategy involving bienterostomyper primam should be further analyzed. The choice of operative intervention reflects multiple variables, including age, physiologic status, institutional resources and surgeon preference based on experience. Primary peritoneal drainage for perforated NEC may help to resuscitate and treat a critically ill patient initially, and in some instances, may be definitive operative intervention. CONCLUSIONS: Relatively rare disease, of unknown etiology and elusive pathogenesis, NEC has initial non-specific symptoms and clinical features that mimic more common surgical diseases. There is considerable controversy regarding which procedure is preferable. Currently, in the absence of rigorous evidence supporting the superiority of one approach over the other, surgical intervention depends mostly on the treating institution or the individual surgeon.

Microsatellite instability in the management of stage II colorectal patients.

BACKGROUND: Up-to-date it is unclear whether stage II colorectal cancer patients should receive adjuvant chemotherapy.The presence of high risk features (T4, CEA 5 ng dl, less than 12 lymph nodes examined) is an indication for Oxaliplatin based treatment. In their absence, there is no consensus, 5 Fluorouracil regimens, or observation only being equally recommended by oncologists. Microsatellite instability is associated with good prognosis in stage II colorectal cancer and also with poor response to 5 Fluorouracil and should be used as a predictive marker. METHODS: We performed a prospective descriptive study on 115 consecutive patients who received surgical resection for colorectal cancer in our clinic during 2011 and 2012 using a risk stratification algorithm based on TNM staging, clinico pathologic and molecular markers. RESULTS: From the 44 stage II colorectal cancer patients, 10 cases were classified as high risk, in 26 cases we performed Immunohistochemical analysis that identified 8 patients with low risk microsatellite instability phenotype, with no indication for adjuvant chemotherapy; 26 intermediate risk patients received 5-FluoroUracil regimens. CONCLUSION: We believe that microsatellite instability testing provides a useful tool in the goal of better characterizing patients with stage II colorectal cancer in matters of risk of recurrence and likelihood of benefit from chemotherapy.