Dietary supplements in lymphedema

Lymphedema is a chronic inflammatory disorder resulting from ineffective fluid uptake by the lymphatic system, and the effects are principally felt in the lower limbs. The condition is said to be primary when caused by genetic mutations and secondary when caused by injuries, infections, or surgery. Lymphedema, a worldwide pathology, does not have an effective therapy so far. Leukotriene B4 has recently been identified as a key molecule in lymphedema pathogenesis. Surgical, nonsurgical, and pharmacological treatments have been proposed; however, they do not cure the disease and only ameliorate the symptoms. Nutrition and nutritional status are extremely important in lymphedema physiopathology. Obesity is a comorbidity that exacerbates the risk for secondary lymphedema and constitutes a negative prognostic factor. Indeed, anti-inflammatory foods and their effects on the inflammatory state and on oxidative stress are now being investigated for their possible therapeutic role in lymphedema. Although no special diet has so far been proven to be very effective, specific dietary tips could help in alleviating the edematous state of patients with lymphedema. A few supplements have been tested for lymphedema treatment. Among them, GARLIVE® containing hydroxytyrosol, hesperidin, spermidine and vitamin A, exhibited promising effects in the animal model. Hydroxytyrosol, a polyphenol from olives, showed anti-inflammatory effects and reduced leukotriene B4 synthesis, thus holding promise as a potential natural candidate for lymphedema treatment

Optimization of long-range PCR protocol to prepare filaggrin exon 3 libraries for PacBio long-read sequencing

BackgroundThe filaggrin (FLG) protein, encoded by the FLG gene, is an intermediate filament-associated protein that plays a crucial role in the terminal stages of human epidermal differentiation. Loss-of-function mutations in the FLG exon 3 have been associated with skin diseases. The identification of causative mutations is challenging, due to the high sequence homology within its exon 3 (12,753 bp), which includes 10 to 12 filaggrin tandem repeats. With this study we aimed to obtain the whole FLG exon 3 sequence through PacBio technology, once 13-kb amplicons have been generated.Methods and resultsFor the preparation of SMRTbell libraries to be sequenced using PacBio technology, we focused on optimizing a 2-step long-range PCR protocol to generate 13-kb amplicons covering the whole FLG exon 3 sequence. The performance of three long-range DNA polymerases was assessed in an attempt to improve the PCR conditions required for the enzymes to function properly. We focused on optimization of the input template DNA concentration and thermocycling parameters to correctly amplify the entire FLG exon 3 sequence, minimizing non-specific amplification.ConclusionsTaken together, our findings suggested that the PrimeSTAR protocol is suitable for producing the amplicons of the 13-kb FLG whole exon 3 to prepare SMRTbell libraries. We suggest that sequencing the generated amplicons may be useful for identifying LoF variants that are causative of the patients' disorders

Dietary supplements for Lipedema

Lipedema is a chronic disease that mostly manifests in females as the abnormal distribution of subcutaneous adipose connective tissue, usually coupled with bruising, pain, and edema. Lipedema molecular pathophysiology is currently not clear, but several studies suggest that genetics and hormonal imbalance participate in lipedema pathogenesis. Women with lipedema present in some cases with elevated body mass index, and the appearance of obesity in addition to lipedema, where the obesity can cause serious health issues as in lipedema-free individuals with obesity, such as diabetes and cardiovascular disorders.  Unlike obesity, lipedema tissue does not respond well to diet or physical exercise alone. Therefore, in this review we discuss the effect of various dietary supplements that, along with diet and physical exercise, cause fat burning and weight loss, and which could potentially be important in the treatment of lipedema. Indeed, an effective fat burner should convert stored fats into energy, mobilize and break down triglycerides in adipocytes, boost metabolism and inhibit lipogenesis. Common ingredients of fat burning supplements are green tea, caffeine, chromium, carnitine, and conjugated linoleic acid. The use of fat burners could act synergistically with a healthy diet and physical exercise for decreasing adipose tissue deposition in patients with lipedema and resolve related health issues. The effects of fat burners in human studies are sometimes contradictory, and further studies should test their effectiveness in treating lipedema

Peripheral Edema: Differential Diagnosis

Peripheral edemas can be generated by multiple causes, local and/or systemic. The difficulties in recognizing the exact nature of the edema and the cause that originates it often lead to erroneous considerations that determine an inappropriate therapeutic approach. In this chapter the various causes that generate peripheral edema are analyzed (systemic: cardiac diastolic dysfunction, kidney failure, liver failure, myxedema, from drugs, and idiopathic; and local: venous and/or lymphatic transport insufficiency). They are also described, according to the diagnosis made and the clinical and instrumental criteria to attain a correct and early diagnosis and to proceed to the most appropriate therapeutic measures (drugs, surgery, physical rehabilitative by means of manual and mechanical techniques) in individual cases

Familial hyperhomocysteinemia, age and peripheral vascular diseases - an Italian study

Hyperhomocysteinemia is a widely recognized, although not yet entirely understood, risk factor for cardiovascular disease. Particularly, the complex relationships between age, hyperhomocysteinemia, predisposing genetic factors and peripheral vascular diseases have not been fully evaluated. Our contribution to this issue is a retrospective analysis of a large series of patients with peripheral arterial, venous and lymphatic disease, and of their blood relatives, with special reference to homocysteine plasma levels, age and methylenetetrahydrofolate reductase (MTHFR) polymorphisms. Serum homocysteine was measured in 477 patients (286 males, 191 females, age range 19-78 years) with various vascular clinical conditions: postphlebitic syndrome (46) recurrent venous ulcers (78), arterial diseases (101) primary lymphoedema (87), secondary lymphoedema (161) and outlet thoracic syndrome (4), and in 50 normal controls. A MTHFR study for polymorphisms was carried on in the subjects with homocysteine values exceeding 15 mol/L. Serum homocysteine determination and MTHFR polymorphism studies were performed also in 1430 healthy blood related relatives (mainly siblings, descendents and sibling descendents) of the subjects with hyperhomocysteinemia and MTHFR polymorphisms. We found MTHFR polymorphisms in 20% of controls and in 69.3%, 69.5% and 53.8% of hyperhomocysteinemic subjects with arterial diseases, postphlebitic syndrome and venous ulcers, respectively. As expected, the percentage of hyperhomocysteinemia in patients with secondary lymphoedema and with thoracic outlet syndrome did not show significant differences compared to the control group. A MTHFR polymorphism was found in 116 out of the 214 hyperhomocysteinemic patients, i.e., in the 54% of the overall patient population with hyperhomocysteinemia (214 patients). Interestingly 750 (52%) out of the 1430 blood relatives of the 116 patients with hyperhomocysteinemia and MTHFR polymorphisms showed at least one polymorphism in MTHFR gene. In this latter group of 750 healthy blood-related relatives bearing a MTHFR polymorphism the finding of hyperhomocysteinemia increased according to the age class from 1.6% in the age range 60 years. The present study demonstrate that patients with peripheral arterial disease, post-phlebitic syndrome, venous ulcers and primary lymphoedema show a significantly higher incidence of hyperhomocysteinemia compared to controls, and adds further evidence to the causative role of hyperhomocysteinemia in the development of both arterial and venous disease. Moreover our data indicate a possible causative role of hyperhomocysteinemia in primary lymphoedema. In more than 50% of our hyperhomocysteinemic patients a polymorphism of MTHFR (C677T and/or A1298C) was detected. In subjects with these polymorphisms the frequency of hyperhomocysteinemia increases with age. We observed a quite similar frequency of the two polymorphisms in the studied population and therefore claim for the need to study both C677T and A1298C mutations in hyperhomocysteinemic patients

Aldo-Keto Reductase 1C1 (AKR1C1) as the first mutated gene in a family with nonsyndromic primary lipedema

Lipedema is an often underdiagnosed chronic disorder that affects subcutaneous adipose tissue almost exclusively in women, which leads to disproportionate fat accumulation in the lower and upper body extremities. Common comorbidities include anxiety, depression, and pain. The correlation between mood disorder and subcutaneous fat deposition suggests the involvement of steroids metabolism and neurohormones signaling, however no clear association has been established so far. In this study, we report on a family with three patients affected by sex-limited autosomal dominant nonsyndromic lipedema. They had been screened by whole exome sequencing (WES) which led to the discovery of a missense variant p.(Leu213Gln) in AKR1C1, the gene encoding for an aldo-keto reductase catalyzing the reduction of progesterone to its inactive form, 20-\u3b1-hydroxyprogesterone. Comparative molecular dynamics simulations of the wild-type vs. variant enzyme, corroborated by a thorough structural and functional bioinformatic analysis, suggest a partial loss-of-function of the variant. This would result in a slower and less efficient reduction of progesterone to hydroxyprogesterone and an increased subcutaneous fat deposition in variant carriers. Overall, our results suggest that AKR1C1 is the first candidate gene associated with nonsyndromic lipedema

The Italian Earthquakes and Tsunami Monitoring and Surveillance Systems

The Osservatorio Nazionale Terremoti (ONT) is the Italian seismic operational centre for monitoring earthquake, it is part of Istituto Nazionale di Geofisica e Vulcanologia (INGV) the largest Italian research institution, with focus in Earth Sciences. INGV runs the Italian National Seismic Network (network code IV) and other networks at national scale for monitoring earthquakes and tsunami. INGV is a primary node of European Integrated Data Archive (EIDA) for archiving and distributing, continuous, quality checked seismic waveforms (strong motion and weak motion recordings). ONT designed the data acquisition system to accomplish, in near-real-time, automatic earthquake detection, hypocentre and magnitude determination and evaluation of moment tensors, shake maps and other products. Database archiving of all parametric results are closely linked to the existing procedures of the INGV seismic monitoring environment and surveillance procedures. ONT organize the Italian earthquake surveillance service and the tsunami alert service (INGV is Tsunami Service Provider of the ICG/NEAM for the entire Mediterranean basin). We provide information to the Dipartimento di Protezione Civile (DPC) and to several Mediterranean countries. Earthquakes information are revised routinely by the analysts of the Italian Seismic Bulletin. The results are published on the web and are available to the scientific community and the general public.PublishedMontreal1SR TERREMOTI - Sorveglianza Sismica e Allerta Tsunam

Ethical considerations regarding animal experimentation

Animal experimentation is widely used around the world for the identification of the root causes of various diseases in humans and animals and for exploring treatment options. Among the several animal species, rats, mice and purpose-bred birds comprise almost 90% of the animals that are used for research purpose. However, growing awareness of the sentience of animals and their experience of pain and suffering has led to strong opposition to animal research among many scientists and the general public. In addition, the usefulness of extrapolating animal data to humans has been questioned. This has led to Ethical Committees’ adoption of the ‘four Rs’ principles (Reduction, Refinement, Replacement and Responsibility) as a guide when making decisions regarding animal experimentation. Some of the essential considerations for humane animal experimentation are presented in this review along with the requirement for investigator training. Due to the ethical issues surrounding the use of animals in experimentation, their use is declining in those research areas where alternative in vitro or in silico methods are available. However, so far it has not been possible to dispense with experimental animals completely and further research is needed to provide a road map to robust alternatives before their use can be fully discontinued

Methodology for clinical research

A clinical research requires a systematic approach with diligent planning, execution and sampling in order to obtain reliable and validated results, as well as an understanding of each research methodology is essential for researchers. Indeed, selecting an inappropriate study type, an error that cannot be corrected after the beginning of a study, results in flawed methodology. The results of clinical research studies enhance the repertoire of knowledge regarding a disease pathogenicity, an existing or newly discovered medication, surgical or diagnostic procedure or medical device. Medical research can be divided into primary and secondary research, where primary research involves conducting studies and collecting raw data, which is then analysed and evaluated in secondary research. The successful deployment of clinical research methodology depends upon several factors. These include the type of study, the objectives, the population, study design, methodology/techniques and the sampling and statistical procedures used. Among the different types of clinical studies, we can recognize descriptive or analytical studies, which can be further categorized in observational and experimental. Finally, also pre-clinical studies are of outmost importance, representing the steppingstone of clinical trials. It is therefore important to understand the types of method for clinical research. Thus, this review focused on various aspects of the methodology and describes the crucial steps of the conceptual and executive stages

SISMIKO:emergency network deployment and data sharing for the 2016 central Italy seismic sequence

At 01:36 UTC (03:36 local time) on August 24th 2016, an earthquake Mw 6.0 struck an extensive sector of the central Apennines (coordinates: latitude 42.70° N, longitude 13.23° E, 8.0 km depth). The earthquake caused about 300 casualties and severe damage to the historical buildings and economic activity in an area located near the borders of the Umbria, Lazio, Abruzzo and Marche regions. The Istituto Nazionale di Geofisica e Vulcanologia (INGV) located in few minutes the hypocenter near Accumoli, a small town in the province of Rieti. In the hours after the quake, dozens of events were recorded by the National Seismic Network (Rete Sismica Nazionale, RSN) of the INGV, many of which had a ML > 3.0. The density and coverage of the RSN in the epicentral area meant the epicenter and magnitude of the main event and subsequent shocks that followed it in the early hours of the seismic sequence were well constrained. However, in order to better constrain the localizations of the aftershock hypocenters, especially the depths, a denser seismic monitoring network was needed. Just after the mainshock, SISMIKO, the coordinating body of the emergency seismic network at INGV, was activated in order to install a temporary seismic network integrated with the existing permanent network in the epicentral area. From August the 24th to the 30th, SISMIKO deployed eighteen seismic stations, generally six components (equipped with both velocimeter and accelerometer), with thirteen of the seismic station transmitting in real-time to the INGV seismic monitoring room in Rome. The design and geometry of the temporary network was decided in consolation with other groups who were deploying seismic stations in the region, namely EMERSITO (a group studying site-effects), and the emergency Italian strong motion network (RAN) managed by the National Civil Protection Department (DPC). Further 25 BB temporary seismic stations were deployed by colleagues of the British Geological Survey (BGS) and the School of Geosciences, University of Edinburgh in collaboration with INGV. All data acquired from SISMIKO stations, are quickly available at the European Integrated Data Archive (EIDA). The data acquired by the SISMIKO stations were included in the preliminary analysis that was performed by the Bollettino Sismico Italiano (BSI), the Centro Nazionale Terremoti (CNT) staff working in Ancona, and the INGV-MI, described below