Most male BC patients received adjuvant endocrine therapy (AET) with tamoxifen (73.5%). AET drop-out rate due to side effects was 16.3% for males compared to 7.6% for women (p = 0.04). Comparing FBC and male BC, no differences have been identified in terms of DFS and OS, with a similar 10-year-relapse rate . Propensity Score Matching by age, nodal status, pT, and molecular subtype had been performed and no differences in OS and DFS were seen between male BC and FBC. In conclusion, male BC and FBC have similar prognostic factors and survival outcomes. The drop-out rate of AET was higher in males, and side effects were the main reason for drug discontinuation.Male breast cancer (BC) represents less than 1% of male tumors. Little is known about male BC characteristics, management, and survival, with many studies based on a small number of cases. Consequently, the treatment of male BC lacks specific guidelines. The aims of the study are to compare male and female breast cancer (FBC) in terms of cancer clinical and anatomopathological features and treatment approach, and to identify differences between male BC and FBC in terms of survival. Patients and methods: Data from 2006 to 2018 were retrospectively acquired. Amounts of 49 males and 680 postmenopausal females with primary non-metastatic BC who underwent breast surgery at Mauriziano Hospital or IRCCS Candiolo  were included. The mean age at diagnosis for male BC was 68.6 years, and males presented a smaller tumor size than women ( Breast cancer (BC) is the most common malignant tumor among women worldwide; many researches have been conducted on this subject. Male breast cancer represents less than 1% of male tumors; in Italy, 500 new cases were recorded in 2018 [Males share with females the principal risk factors of developing BC such as age, smoking, and alcohol consumption; in addition, all the causes of hyperestrogenism as cirrhosis, obesity, testicular injuries, or Klinefelter syndrome can increase the risk of BC in males ,3. InherData in the literature are conflicting: in most studies, there are no differences between males and females in terms of prognosis . ConversMale BC usually presents as a unilateral breast lump and undergoes mastectomy in most cases .Lacking specific treatment guidelines and indications for male BC, adjuvant treatments are the same as those used in female breast cancer (FBC). This practice may be suboptimal in some cases considering that hormonal differences between men and women may influence the effectiveness of treatments. For this reason, some aspects of male BC need improvement in specific management, such as surgery, radiotherapy, and systemic treatments ,12.This study aims to compare male and female BC in a cohort of real-life patients, analyzing clinical and anatomopathological features of the tumors and the treatment approach. The goal is to identify differences and possible implications to improve disease management and survival.We retrospectively compared 49 male patients to 680 female postmenopausal patients with a minimum follow-up available of 36 months with primary non-metastatic BC. All the patients underwent breast surgery at the Academic Department of Obstetrics and Gynaecology of “Umberto I” Hospital in Torino and IRCCS Hospital in Candiolo between 1 January 2006 and 31 December 2018.Data regarding the patient’s medical history, age, ongoing therapies, previous surgery, personal and hereditary oncological history, BRCA1/2 gene mutation, clinical and biological characteristics of the tumor, and follow-up data were obtained from prospectively maintained, non-open-access institutional databases.As recommended by the most recent version of the NCCN guideline, all the specimens had been tested for hormone receptors such as estrogen (ER) and progesterone receptors (PgR) by immunohistochemistry (IHC). Cancers were considered hormone-receptor-positive if at least 1% of the cells tested expressed ER. Otherwise, the tumor was considered hormone receptor-negative ,14,15. HWe classified both male and female BC using IHC surrogates for molecular subtypes, as follows: luminal A , luminal B , HER-2 (ER and PgR negative and HER-2 positive) and basal-like  ,18.Surgical and adjuvant medical treatment decisions were based on current international guidelines at the time of surgery ,20,21. PConsent to the anonymous use of their clinical and instrumental data for scientific purposes is routinely signed by all patients treated at both institutes.The Kolmorogov–Smirnov test was performed to evaluate the normal distribution of the variables. The association and the statistical significance of the categorical variables have been verified with Chi-square or Fisher’s exact test, when applicable. The t-Student test has been used to compare continuous variables with normal distribution.The survival endpoints were Disease Free Survival (DFS) and Overall Survival (OS) and Kaplan–Meier curves and log-rank tests were carried out in the two groups. Univariate and multivariate Cox regression analyses were also performed for survival endpoints and hazard ratios (HR) with 95% confidence intervals were reported.To reduce the bias due to the effects of confounding variables in the two cohorts, Propensity Score Matching (PSM) has been performed for DFS and OS. The patients have been matched 1:1 by age at diagnosis, nodal status, pT, and molecular subtype so 49 males have been matched with 49 women. DFS and OS have been calculated.p-values < 0.05 were considered to be statistically significant.Data were analyzed using SPSS software, version 25.0 for macOS .The characteristics of the entire population, male and female, are presented in The mean age at diagnosis for male BC was 68.6 years (±10.1). The first symptom was, in most of the male BC cases, the presence of a unilateral breast lump (73.5% of the patients); less often nipple discharge or nipple retraction (26.5% of the patients).Six males carried genetic mutations: four patients had a BRCA2 mutation, one patient a CHECK2 mutation, and one patient presented MLH1 mutation (associated with Lynch syndrome).p < 0.05) than women. In both populations, tumors were pT1 or pT2 in more than half of the cases. An amount of 14% of male BC were diagnosed at pT4 stage compared with only 6% of women (p = 0.01). In contrast, no men with pT3 tumors were present in our case series.Males presented a smaller tumor size . Axillary surgery was not performed in 25 women and in a man due to poor general conditions.Males and females presented node-negative disease in 67.3% and 64.7%, respectively (p < 0.05). Nearly 90% of male BC were IHC luminal A or B subtypes, and we had no cases of basal-like male BC . According to IHC classification, male BC was more frequently luminal B than FBC .The most common histological features in male BC were the non-special type (NST) ductal carcinoma subtype with high nuclear grade (G3) and elevated Ki67. A lower rate of lobular tumors was noticed among male BC compared with females . Axillary surgery was carried out following the indications given by national and international guidelines. Specifically, most of our male patients (n = 33) presented clinically N0 at diagnosis, and BLNS was performed, while ten patients had lymph node involvement, so they underwent ALND. In 5 cases, axillary dissection was performed following the finding of lymph node metastasis at BLNS in patients who underwent a mastectomy. In one case, axillary surgery was omitted due to the patient’s poor general clinical condition.A relevant difference between male BC and FBC is the type of surgery performed. In fact if all men with BC underwent mastectomy, this type of surgery was performed in only one third of the women (p < 0.05). AET dropout rate due to side effects was 16.3% for males compared to 7.6% for women (p = 0.04).Most male BC patients received adjuvant endocrine therapy (AET) with tamoxifen (73.5%). Six patients received no AET due to personal refusal. In agreement with the initial selection criteria , the majority of FBC patients had AET with aromatase inhibitors (p < 0.05), probably due to the higher rate of pT4 tumors diagnosed in males.Radiotherapy on the chest wall was indicated in 16.3% of male BC compared with 4.1% of FBC .Univariate analysis for DFS performed on the entire female population identified the following risk factors for recurrence: nodal involvement, G3, tumoral diameter > 2 cm, Ki67 > 20%, and luminal B-type and basal-like IHC subtypes. The drop-out of AET had no apparent impact on DFS .Univariate analysis for DFS in male BC showed prognostically unfavorable factors tumor dimension > 20 mm, nodal involvement, G3, and luminal B IHC subtype. .In male BC, multivariate Cox regression analysis for DFS confirmed tumoral dimensions > 20 mm, nodal involvement, and G3 as independent negative prognostic factors in male BC  Figure . In FBC,OS data of FBC obtained from univariate analysis showed that pT3, histologic grade ≥ 2, and basal-like tumors resulted in reduced survival . Focusinp = 0.3) and OS (p = 0.34)  , with a p = 0.43) and in DFS (p = 0.09) were seen in the two groups , and the most frequent IHC subtype is luminal, whereas basal-like and HER2+ BC are less represented  in agreement with Johansson et al. and a SEER database analysis published in 2012 and other studies ,29,30.Regarding surgical treatment, all males underwent mastectomy compared with 36% of women. The twofold likelihood of males receiving mastectomy is probably due to the mammary gland tumor ratio and the higher frequency of pT4 tumors, thus making mastectomy the most appropriate surgery in accordance with guidelines .In line with the literature, endocrine therapy plays a leading role among systemic adjuvant treatments in hormone-responsive tumors, in both males and females, reducing the rate of recurrence and increasing survival . Recent In men, tamoxifen represents the most employed drug, followed by aromatase inhibitors associated or not with ovarian suppression function (OSF) with GnRH agonists ,33.Besides the evident benefits, AET is burdened with non-negligible side effects that may lead to drug discontinuation. As mentioned in a recent paper published by our group, the main side effects that can lead to drug drop-outs in up to 11.3% of patients taking AI and 13.8% of premenopausal patients taking tamoxifen ±OFS are related to anti-estrogen action such as hot flashes, vaginal dryness, sexual dysfunction, and dyspareunia, increased thrombotic risk, weight gain, arthralgias, and insomnia .In the present study, AET was well tolerated by women with a dropout rate of less than 10%, probably due to the inclusion of only postmenopausal women. On the contrary, the adherence to AET was statistically lower for males with a discontinuation rate of 16% due to the side effects cited above, in particular decreased libido, weight gain, and hot flashes. Interestingly, such discontinuation had no impact on the risk of recurrence. This finding needs more data to be confirmed but could reflect tumor heterogeneity and complexity. In this regard, several trials are underway evaluating genomic differences between FBC and male BC, and testing the efficacy of drugs directed against the androgen receptor .Forty percent of both women and men received adjuvant chemotherapy. The literature has not demonstrated a significant difference between men and women concerning the use of chemotherapy for BC. A recent study from Giordano et al. including 135 patients with a median follow-up of 14 years, pointed out that adjuvant chemotherapy  reduces male BC mortality by 43% and also significantly reduces the risk of recurrence .Data on male BC survival outcomes widely vary in the literature and are often based on low-number studies; on the contrary, studies that include a higher number of patients date back many years when there were no screening programs and treatment options were different .Taking into account recent data, it has been shown that the cumulative 5-year survival rate of male BC is between 36 and 66% and in some studies appears statistically lower than the survival rate in women (80–86%), mainly due to the more advanced stage at presentation of male BC ,38. In cIn our study, based on a real-life cohort of female and male patients, the prognostic factors for worse DFS were equal for both sexes. In females, the main factors related to recurrence are tumoral diameter > 20 mm and nodal involvement; similarly, in males, the tumor characteristics significantly and independently correlated with worse DFS were tumor diameter > 20 mm, nodal involvement, and G3.In our population, no significant differences in survival and risk of recurrence at ten years between males and females had been seen, with an almost similar 10-year relapse rate in the two groups, as well as in other studies ,43,44.To eliminate confounding factors and selection bias, we applied the PSM by pairing females with males. In contrast to Scomersi et al. who obtained a higher risk of recurrence in male BC after PSM, we observed no differences in DFS and OS in our case series between females and males . This reTo the best of our knowledge, ours is one of the few real-life studies in the literature with a discrete sample size of male BC and an average follow-up of more than 5 years. Furthermore, our patients were treated by the same surgical team and with the same medical and surgical treatment protocols. However, studies with a larger sample size are needed to confirm our data.The future perspective of our study is to expand the case series of males and evaluate the impact of BRCA1/2 mutation on clinicopathological features and survival outcomes.According to our preliminary data, we can assume that male BC and FBC have similar prognostic factors and survival outcomes. The drop-out rate of AET was higher in males and the side effects were the main reason for drug discontinuation; the real challenge will be to identify male BC characteristics and develop more specific and tolerable therapies.Like FBC, male BC presents great complexity and heterogeneity. Further randomized prospective clinical trials are needed to clarify the biology and molecular pathways to achieve more targeted and tailored treatments.


Thus, such colloids possess giant dielectric permittivity which shows peculiar temperature dependence, as well as quasi-periodic fluctuations during slow evaporation or dilution. In these colloids, DND interacts with water and air to form cottonwool-like fibers comprising living micro-organisms (fungi and bacteria) and DND particles, with elevated nitrogen content due to fixation of atmospheric N2. Prolonged contact between these solutions and air lead to the formation of ammonium nitrate, sometimes forming macroscopic crystals. The latter was also formed during prolonged oxidation of fungi in aqueous DND colloids. The possible mechanism of N2 fixation is discussed, which can be attributable to the high reactivity of DND. Aqueous suspensions (colloids) containing detonation nano-diamond (DND) feature in most applications of DND and are an indispensable stage of its production; therefore, the interaction of DND with water is actively studied. However, insufficient attention has been paid to the unique physico-chemical and biological properties of transparent colloids with  The synthesis of nanoscale diamond powder by detonating explosives in the absence of air is a unique example of when, as a result of fundamental research, the expensive and environmentally harmful disposal of explosives after the expiration of their safe storage time (by incineration or dissolving in water) has been turned into a profitable industrial production of nanomaterials with huge prospects for their use in science, various technologies, and for biomedical purposes. DNDs, with typical crystal sizes of 4–5 nm, are produced by detonating high explosives enclosed in an ice shell . The latThe first detonation synthesis of nanodiamond published in ,2 initia4, the suspension formed by using sonication and subsequent centrifugation at 15,000 rpm also was converted into a clear colorless suspension containing ca. 0.01–0.1 wt% of DND, henceforth referred to as ‘diamond water’ (DW). This liquid displays a variety of unique and puzzling properties, some of which hold technological promise; please refer to [Aqueous DND suspensions with various carbon contents (wt%): 0.007–0.2 , 0.05 7,9, 0.6–3refer to  for addi3 (diamond-like) carbon. The surface, however, cannot consist of the latter without allowing ‘dangling’ bonds and is either reconstructed into well-ordered chains of π-bonded carbon atoms  [carbon, typically containing 10–15% of other elements. Colloidal suspensions are complex systems. This complexity depends on the materials, their concentrations, the particle sizes, and various atomic interactions, such as electrostatic repulsion, depletion forces, van der Waals attraction and Brownian motion, with the coagulation of particles being another important effect. Furthermore, a DND particle itself has a complex, variable and still-debated structure  15,16,1,18 and sruction) , convertruction)  or is teSuspensions with a high carbon content are typical heterogeneous systems, containing a mixture of liquid and solid phases, which, after intensive centrifugation, turn into homogeneous transparent (to the human eye) solutions containing particles less than 1 μm in size. Indeed, studies of these liquids by the dynamic light scattering (DLS) method show that the particle-size distribution in them varies from tens to hundreds of nanometers ,8,12,21.2g−1 from Brunauer–Emmett–Teller (BET) adsorption–desorption data, which corresponds to effective particle sizes of 6–10 nm, almost as small as the primary particles [However, this agglomeration is not just a close packing of primary particles, as was presumed initially. Firstly, agglomerated DND has specific surface area of 180 to 300 marticles ,5. Seconarticles  or at learticles  as the particles . The conarticles ), and DNarticles . These farticles ,20 by prIn the examination of dry DND powder using static light scattering (SLS) as depicted in n = 2.1 in dry DND), which corresponds to the 14% water content within these particles, and the density of particles (ρp) in DW equaled 2.2 g/cm3. This fact is consistent with the strength of bonding between water molecules and the surface of DND [It was shown by Fabry–Perot interferometry  that cole of DND  and the e of DND ,24.D) of DND particles in water, their corresponding volumes (Vp) and the concentrations (C) of two samples, calculated as 0.01/2.2 = 0.045 and 0.1/2.2 = 0.45 vol% (wt%/ρp). Additionally, the table presents the water volume per particle (Vw = Vp) and the radii of water spheres (Rw) surrounding DND particles within DWs ectively ,23,29, lectively ,12, chemectively  and the ectively .τprec) of a particle isD is the particle size, ρ is its density, h is the height of its descent, η and ρo are the viscosity and density of the medium  and g is the gravitational acceleration. Equation (1) predicts that DND agglomerates with D = 1, 0.5 and 0.1 μm would precipitate in 4 h, 16 h and 17 days, respectively, while 10 μm ones in just 2.5 min. On the contrary, primary DND particles of 5 or 10 nm, if dispersed, will remain suspended practically indefinitely , although their density is higher than that of agglomerates, which are a mixture of porous DND frameworks and water. This, of course, is a crude estimate, precipitation being a complicated process. To the authors’ knowledge, no direct experimental measurement of this interesting effect has been reported so far; however, it is noteworthy that hydrosols of deagglomerated primary DND particles proved remarkably stable and did not show any turbidity or precipitation of solid over several years , with relatively low dielectric permittivity , shows similar behavior.DW is slightly acidic. Remarkably, its pH of 6.25–6.77 does not change after long exposure to atmospheric air, i.e., to atmospheric COpolar organic solvents are higher than those of the solvents themselves, increasing sympathetically with the dipole moment of the solvent, as shown in The transparent colloidal solutions of DND are the most convenient objects for electro-physical measurements, which are very sensitive to changes in the atomic and electronic structure of homogeneous media. For example, in , it was In DWs, the effect is much greater and shows the trend of change in the dielectric constant (ε) and the resistivity (R) with the concentration of DND, as shown in 4 to 106 at AC of 1 kHz (cf. 78 for pure water). Although dry DND has higher ε , which is very difficult to avoid, amplifies the effect. Obviously, the same can be expected from DND particles prepared under oxidative conditions, which has ionizable surface-terminating groups (carboxylic and hydroxylic). However, electrode polarization effects only became noticeable at frequencies lower than 1 kHz; above this point, the curve was practically flat [Notwithstanding the low concentration of DND, the colloids showed giant ε, varying for different preparations from 10d ε = 21 ) than bud ε = 21 ,34; corrd ε = 21 . This aplly flat ,34. TherNumerous studies of electro-physical properties in hydrated powders, aqueous suspensions and colloids ,42,43,44An obvious question is whether the colloid remains spatially homogenous during (and after) freezing. Indeed, on slow (48 h) freezing of DW, DND particles were observed to concentrate in one end of the frozen sample . HThe fluorescent properties of DND make it possible to monitor the distribution of particles in DW in real time. increasing changes of NMR relaxation times in highly diluted aqueous solutions [According to standard DLVO theory, interfacial effects occur at distances of a few nm from surfaces of solids , but theolutions  also sug8 to 1010-fold dilution, ε(DW) approached that of pure bulk water. Also, somewhat paradoxically, the first three dilutions  caused successive increases of the DND particle sizes spread (according to DLS measurements), as Non-monotonic changes of permittivity and conductivity of DW occur not only during its evaporation , but alslower than that of bulk water (BW). If this is so and the structure  of the solid component remain unaltered, then where does the giant DP of the water–solid combination comes from? According to theoretical models [The above-mentioned new experimental data, along with previously known facts ,51,52,53l models ,70,71,72l models ,65, to 7l models  in the pl models  that CW 5–10−6 (at 1 kHz) at room temperature and experiences a significant increase upon heating. This effect is one of greatest complexity and still imperfectly understood; however, we always observed the direct dependence of permittivity on conductivity of DW, according to the known relationnot at the freezing point. The observed values of ε(DW) are quite unprecedented. Structural changes in aqueous colloids are detectable due to very high sensitivity of the dielectric method: in DW with carbon content within hundredths of one per cent, ε reaches 10articles . DW haviEp and Eb are the atomization energies, and Np t and Nb are the average coordination numbers of atoms in the particle and in the bulk, respectively. The Ep calculated by Equation (3) for some elemental solids  formed and gradually increased in size, ultimately reaching several mm in length. In identical vials of pure water kept alongside, no fibers were formed; moreover, if transplanted there, the fibers failed to develop and soon disintegrated . AuthorsFurther DWF studies showed that its content changes under different treatments, see 2 molecule into ammonia and/or nitrate) occurs. The nitrogen content increases with the duration of exposure DW to air; it also increases when air is replaced by pure N2 gas. These facts speak in favor of nitrogen fixation from the air. Finally, a replacement of DND with nitrogen-free synthetic diamond (AS) prepared under static high pressure . The slow kinetics of these reactions is explained by the same low concentration of nanodiamond, which is a carrier of fixed nitrogen from air to water. A very small concentration of these particles, which served as centers of crystallization of the formed NH2 molecules in the atmosphere. The ‘fixation’ of atmospheric nitrogen, which involves breaking up the extremely strong N≡N bond, and its conversion into chemically useable compounds, e.g., ammonia or nitrogen oxides, is thus a major challenge in chemistry. The importance of fixation, which has been carried out by the Haber–Bosch method for more than 100 years, is evident from the huge scale of global ammonia production (170 million tons per year), which accounts for up to 1–2% of the global energy consumption. The high cost, high energy costs and environmental harm (the release of a huge amount of carbon dioxide into the atmosphere) of this method make us look for alternatives, to which up to 30,000 publications and citations for the topic of ammonia synthesis have been devoted annually [It is well known that nitrogen is vital for all living organisms, as an ingredient of proteins and nucleic acids. It is also necessary for production of various industrial chemicals, both large-scale and fine. However, most of the nitrogen on Earth is in the form of chemically inert Nannually . encouraging cellular  growth has not attracted attention it deserves, given the practical applications of DND in medicine .Bacterial and fungal spores are omnipresent in air and infestation of water exposed to it is only a matter of time; however, we have shown that an admixture of nanodiamond, even in small concentration, greatly facilitates such infestation and subsequent growth in bacterial and fungal colonies. The effects of DND on living cells have been studied extensively, but usually with the focus on biocompatibility, i.e., the absence of cytotoxicity. It has been shown that DND is biocompatible with a broad variety of eukaryotic cells ,97,98, bAs mentioned above, in vials of DW exposed to atmospheric air at room temperature, after some periods of time, cloudy and then cotton wool-like fibers (DWF) were observed forming. In identical vials of pure water kept alongside, no fibers were formed; moreover, if transplanted there, the fibers failed to develop and soon disintegrated. The IR spectra of DWFs were very similar to those of typical fungi . BiochemPenicillium cyclopium, P. aurantiogriseum, P. aurantiogriseum, P. chrysogenum, Purpureocillium lilacinum, Beaveria bassiana, some species of the genera Mucor, Rhizopus, Trichoderma and Aspergillus, as well as some unidentified species. Various bacterial mucous colonies were observed, as well as colonies characteristic of Bacillus mycoides, without full identification.Both SEM and optical microscopy  showed DPenicillium bacteria, particularly P. cyclopium, clear white mycelium, and B. mycoides; on the third week, fungal colonies evolved, distinguished by clear dark-colored mycelium.Inoculation with the original DND powder did not produce the growth and formation of colonies of fungi or bacteria after 10 days, even though individual bacteria and fragments of mycelia were present. Inoculation with original DND exposed to air and moisture for a short time, already after 1 week yielded isolated colonies of Neither fungi nor bacteria were isolated using DND previously sterilized in an autoclave (at 1 atm and 394 K), whether immediately before the tests or after kept hermetically sealed for 1 year. Even after a protracted (1 month) incubation of sterile DND in sterile water, fungi or bacteria did not emerge or grow. If the original DND was placed into sterile distilled water, 1 month later some isolated fungal hyphae and bacterial cells were detected. If a sterile DND powder was incubated in ordinary distilled water or a distilled water previously exposed (for 1 to 3 h) to atmospheric air, then the growth in fungal mycelia and bacterial cells was also observed. Note that blank experiments (pure water samples kept next to DW under the same conditions) never produced DWF, and that mature DWF quickly dies if transplanted from DW to pure water. The elemental analysis of the evolving DWF  showed a wide variation of their chemical composition : C, 40 to 74, N, 1.5 to 2.9, H, 1.9 to 3.4, the balance (presumably O), 21.7 to 56.5. The absolute and relative content of carbon and nitrogen in DWF samples generally agrees with that in fungal mycelia and spores, or in bacteria. In micromycetic mycelia the nitrogen content varies from 1.3% to 11% . With thgrowing fibers slowly decreased from 25 to 11 in the first 10 weeks of incubation (cf. 40 in pure DND), then dropped from 4 to <1 after the 12th week. The latter stage is probably the lysis of micro-organisms, whereupon carbon is released as CO2 and nitrogen as ammonia or nitrates. The evolution of the DWF composition is instructive see . The C/N2 gas through DW. Remarkably, N2-bubbling for 24 h gave a lower C/N ratio (10.7) than air bubbling under identical conditions (14.5). Mycelial fungi are known which can develop not just under anaerobic, but even under anaerobic and oligotrophic conditions [The growth in fungi and formation of fibrous and/or fluffy DWF was observed also after bubbling Nnditions ,105. HowAlongside the fibers, we observed peculiar cylindrical features resembling hollow tubes sealed on both ends, the nature of which was not established. X-ray spectral analyses gave the composition of the tubular features as C 64.7 and O 16.6, cf. C 75.8 and O 4.2 wt% for the surrounding DWFs, or C 86.3 and O 4.7 wt% for initial DND powder particles. No other elements with Z > 5 was detected; although the sum total of elements found was below 100%, this can be largely a spurious effect due to the porosity of the samples and lack of a flat horizontal surface. The highest oxygen content of up to 19.3 wt% was found in one of the ‘tubes’ (15 µm wide and 120 µm long). As soon as the electron beam was directed at such a tube, a red flash  appeared inside it and quickly propagated along the entire tube, converting it into ash , i.e., combustion in vacuum occurred, which was remarkable. Similar reaction was observed in DND, where the presence of oxygen makes it a micro-explosive. Indeed, compression of DND powder by Bridgman anvils caused a spontaneous detonation when the pressure reached 23–25 kbar, the smoke produced having the characteristic smell of nitric oxides . The experimental study of the physical, chemical and biological behaviors of clear diluted aqueous colloids of DND led to unexpected results which cannot be explained by known theoretical models.Thus, although the giant DP of confined water on other solids is known, a sharp increase in the DP on heating of DND colloids, and a simultaneous increase in the particle sizes, is completely unexpected. This result rules out the orientation polarization as the cause of giant DP of the solution, since re-orientation of larger DND particles is more difficult. The lattice of the DND particles in aqueous solutions, directly observed in a luminescent microscope due to the fluorescence of nanodiamond grains, indicates inter-particle distances in µm range, which are consistent with the distances calculated from the concentrations of DND in DW. The largest particles in these solutions eventually precipitate due to Stokes’ law, that leads to the restructuring of lattices, i.e., to phase transitions in colloids and changes in their electro-physical properties. 8–1010 times, but is completely absent in non-polar solvents, e.g., CCl4.Giant DP of the DND colloids occurs only in polar solvents and persists in aqueous colloids even after dilution by 102 into an exothermic process, allowing the fixation of atmospheric nitrogen, yielding crystalline NH4NO3 or sustaining bacterial and fungal growth in DND colloids. These results are very important for the ongoing search for new methods of fixation of atmospheric nitrogen under mild conditions, to replace the energy-intensive Haber–Bosch process. The catalytic properties of DND can thus be harnessed to reduce the energy footprint of our civilization while helping to feed the Earth’s growing population. The increased chemical activity of nanosized diamond converts the reaction of carbon with NThis issue is also relevant to one of the most fundamental, as well as fascinating, scientific problems: the origin of life on Earth. The importance of water/solid surfaces was first considered by Bernal , and thi2O, N2, CO, CO2, and smaller amounts of H2, O2, NO, NO2, and NH3 molecules. Therefore, the probability of just five C2 molecules meeting to form a 10-atom diamond cluster is 1/59, i.e., practically nil [6N6O6) as the explosive, thus excluding hydrogen (as well as water and ammonia) from the detonation cloud, the size of the DND diamond core increased fivefold [The peculiarities of physico-chemical and biological properties of DND, reviewed above, are principally due to size effect. Evidently, the smaller the nanodiamond particles, the more manifest this effect would be. However, the observed size distributions in both the dry state and in water, as discussed above, show that in as-produced DND, the particles with the size of ≤5 nm are naturally scarce, which significantly reduces its usefulness for research and technological applications. To reduce the particles sizes, or to increase the proportion of de-agglomerated primary particles, three routes have been suggested, viz., (a) milling with balls (beads) of ever-smaller sizes, to maximize the pressure on impact ; (b) remally nil . On the fivefold .Development of an effective technique for obtaining and preserving de-aggregated primary DND particles of the smallest size would be crucial for the technological prospects of clear DND colloids in physical, chemical and biological applications.


Betahistine was registered in Europe in the 1970s and approved in more than 80 countries as a first-line treatment for Menière's disease. It has been administered to more than 150 million patients. However, according to a Cochrane systematic review of betahistine and recent meta-analyses, there is insufficient evidence to say whether betahistine has any effect in the currently approved dosages of up to 48 mg/d. A combination with the monoamine oxidase B (MAO-B) inhibitor, selegiline, may increase the bioavailability of betahistine to levels similar to the well-established combination of L-DOPA with carbidopa or benserazide in the treatment of Parkinson's disease. We investigated the effect of selegiline on betahistine pharmacokinetics and the safety of the combination in humans.ClinicalTrials.gov.In an investigator-initiated prospective, non-randomized, single-sequence, two-period titration, open label single-center phase 1 study, 15 healthy volunteers received three single oral dosages of betahistine  without and with selegiline (5 mg/d with a loading period of 7 days). Betahistine serum concentrations were measured over a period of 240 min at eight time points . This trial is registered with EudraCT (2019-002610-39) and *ng/mL and for betahistine plus selegiline 53.28 (+/-37.49) h*ng/mL. The half-life time of around 30 min was largely unaffected, except for the 24 mg betahistine dosage. In total, 14 mild adverse events were documented.In all three single betahistine dosages, selegiline increased the betahistine bioavailability about 80- to 100-fold. For instance, the mean (±SD) of the area under curve for betahistine 48 mg alone was 0.64 (+/-0.47) hThis phase 1 trial shows that the MAO-B inhibitor selegiline increases betahistine bioavailability by a factor of about 80 to 100. No safety concerns were detected. Whether the increased bioavailability has an impact on the preventive treatment of Menière's disease, acute vestibular syndrome, or post-BPPV residual dizziness has to be evaluated in placebo-controlled trials.https://clinicaltrials.gov/study/NCT05938517?intr=betahistine%20and%20selegiline&rank=1, identifier: NCT05938517. Betahistine is a structural analog of histamine with weak histamine H1 receptor agonist and more potent histamine H3 receptor antagonist properties. Betahistine has been used for various indications, namely for the prophylactic treatment of Menière's disease for over 50 years and for the symptomatic treatment of vertigo. It is approved for Menière's disease in more than 80 countries and has been used in over 130 million patients with a favorable safety profile ; this fiA basic pharmacokinetic problem of betahistine is that 99% of orally ingested betahistine is metabolized in the gastrointestinal tract and liver by MAO-B/A . This fiIn an animal model of unilateral vestibular neurectomized cats, oral treatment with betahistine in combination with selegiline led to significantly higher blood concentrations of betahistine and to a better posture function , 13. A sBased on these findings, the primary objective of this phase 1 trial in healthy subjects was to demonstrate that the serum concentration of betahistine is higher in a combination treatment with selegiline compared to betahistine monotherapy. The primary endpoint was the area under the curve (AUC0-240 min) of the serum concentrations of betahistine after combination treatment with betahistine and 5 mg/d selegiline compared to betahistine monotherapy, giving single dosages of 24, 48, and 96 mg betahistine in ascending dosage. The secondary objectives were the assessment of safety of the combination treatment and the pharmacokinetics of betahistine in different dosages in serum, namely the half-life time and the pharmacokinetic simulation of multiple dosing.This study was an investigator-initiated (IIT), prospective, non-randomized, single sequence, two-period titration, open label single-center phase 1 study in healthy volunteers. It was conducted at the Department of Neurology, LMU University Hospital, LMU Munich, Munich, Germany with recruitment between 2 June 2021 and 9 September 2021.ClinicalTrials.gov.The study was performed in compliance with the requirements of the “Bundesinstitut für Arzneimittel und Medizinprodukte” (BfArM), Fachregistratur Klinische Pruefung, Kurt-Georg-Kiesinger-Allee 3, 53175 Bonn and Regierung von Oberbayern, Sachgebiet 55.2, 80534 Munich. It gained full regulatory approval from the Bundesinstitut für Arzneimittel und Medizinprodukte on 05.06.2020. The study was approved by the local ethics committees in Munich (19-992) on 25.05.2020. In the EudraCT register, the study was issued with the EudraCT number 2019-002610-39, registered on 17 June 2019 (first patient in 2 June 2021), named PK-BeST. As phase 1 studies are not public in the EudraCT register, the study was also registered in Subjects were recruited via an announcement on the clinic homepage/intranet and were adult (age ≥18 years and ≤ 70 years) healthy volunteers  as determined by screening and the principal investigator's judgement. Participants were not allowed to take medication due to an illness and had normal blood pressure, heart rate, and ECG values.Exclusion criteria were conditions in which repeated serum draws or injections pose more than minimal risk for the subjects, such as hemophilia, a positive test for alcohol or drugs, known contraindications for betahistine, such as bronchial asthma, and known contraindications for selegiline, e.g., hypersensitivity.Each subject gave their written informed consent before any study-related procedures were performed.Study visits took place in an outpatient setting. The first step was to check the inclusion criteria at screening.Subjects received single dosages of betahistine orally  with an at least 48-h wash-out period between the different dosages. In the second period, subjects were pre-treated with selegiline orally (5 mg/day) for 1 week and treated continuously with selegiline (5 mg/day) in combination with the single dosages of betahistine  with at least 2 days between the different dosages.After absorption, betahistine is rapidly and almost completely metabolized into 2-PAA . After oral administration of betahistine, the plasma (and urinary) concentration of 2-PAA reaches its maximum 1 h after intake and declines with a half-life of about 3.5 h. A wash-out-time of 48 h, i.e., 15 times the half-life time, is sufficient and was feasible.Vital signs and adverse events were checked for a minimum of 4 h after the intake of betahistine in the center. Serum samples to test serum concentrations of betahistine were obtained before and 10, 30, 60, 90, 120, 180, and 240 min after betahistine intake.Safety parameters included assessment of adverse events, ECG, vital signs, laboratory measurements including kidney and liver function, full blood count, and pregnancy and drug screening test. Safety monitoring was conducted during every visit and assessment of vital parameters and adverse events was conducted 10, 30, 60, 90, 120, 180, and 240 min after betahistine intake. The clinical trial was conducted in a hospital of maximum care with the possibility to consult a specialist for possible symptoms of intoxication, specifically an anesthesiologist, cardiologist, and neurologist 24 h/7 days a week.This was a phase 1 open label study. All subjects received the same treatment and no blinding process was involved. Due to safety reasons, ascending dosages were required by the federal institute so that no randomization was possible.The primary endpoint was the area under the curve (AUC0-240 min) of the serum concentrations of betahistine after combination treatment with betahistine and 5 mg/d selegiline compared to betahistine monotherapy, giving single dosages of betahistine of 24, 48, and 96 mg in ascending dosage. Secondary endpoints were the half-life of betahistine in serum after intake of different dosages of betahistine and the occurrence of adverse effects due to combination treatment with betahistine and selegiline.post-hoc to evaluate a possible accumulation over time. The simulations of multiple dosing regimens were conducted using a one-compartment model with PKTool, a free pharmacokinetic prediction tool developed with funding from the Bill & Melinda Gates Foundation . As certain PK parameters necessary for simulating multiple dosing are unknown from empirical studies , estimates were obtained by fitting values to a one-compartment model using the single dose data with PKsolver (Computer simulations of multiple dosing with the three different dosages of betahistine and dosing regimens (three times per day and four times per day) were performed PKsolver .The sample size chosen for this study was not based on statistical considerations. The number of subjects per dosage was chosen based on historical experience with safety and tolerance trials. The sample size falls within the range of those used in other studies of this nature. It was planned to enroll a total number of 15 subjects between 18 and 70 years of age in this trial at one trial site.Our preliminary analysis includes descriptive statistics of demographics, general physical examination, relevant medical history, and concomitant medication and neurological examinations. Numerical variables  are summarized in terms of mean (SD), median (minimum-maximum), and median . Categorical variables  are summarized in terms of frequency counts and percentages.For our study, extensive graphical analyses preceded every population model analysis. Concentration-time data was plotted by individuals . Plots of the summarized data are also presented.Tmaxax, and Cmaxax), summary statistics are presented in a table followed by scatter plots on a linear y-axis and a log y-axis and bar charts. This is followed by the normality tests for linear and log transformed data.For pharmacokinetic parameters  of the serum concentrations of betahistine after combination treatment with betahistine and 5 mg/d selegiline compared to betahistine monotherapy, giving single dosages of betahistine of 24, 48, and 96 mg in ascending order. The efficacy of the experimental intervention was determined by AUC0-240 min of betahistine before and 10, 30, 60, 90, 120, 180, and 240 min after treatment in the two treatment arms.Tmaxax, and Cmaxax). The values from each individual were then used to calculate a mean and standard error of the mean.All data were plotted as grouped  and as individual data. The graphs of each individual's serum betahistine concentration over time was used to detect possible errant data points. The data from individual plots were then used to calculate the pharmacokinetic parameters .Log10 data.For statistical analysis, as all pharmacokinetic parameters seemed to be not normally distributed, the data were log transformed. There was no evidence that the data were not log normal. All statistical tests were performed on the Log10 transformed AUC0-240 min values. A repeated measure ANOVA was used with each participant used as a blocking factor . To determine differences between means, rather than comparing all possible means, pre-planned comparisons (“post hoc tests”) were performed to determine the effect of selegiline on pharmacokinetics.Analysis of the primary endpoint was performed using repeated measures ANOVA on the This study was funded by Cures within Reach (CwR), Chicago, USA. This trial was not industry co-sponsored. The funder of the trial had no role in the development of the study design, data collection, data analyses, data interpretation, or the writing of this report. All authors had full access to all data in the study and all authors had final responsibility for the decision to submit for publication.Twenty healthy volunteers were screened for this study between June 2, 2021 and September 9, 2021. Five subjects failed to enter the study for different reasons, mostly laboratory findings or abnormal ECG values. Fifteen participants were enrolled. Of these, all completed the trial and were included in the analysis. For details, see The baseline characteristics of the intention-to-treat sample include demographics, a general physical examination, relevant medical history, and concomitant medication and neurological examinations and are summarized in The mean AUC of betahistine seems to increase with the increased dose of administered betahistine and, more prominently, with the administration of a combination treatment of betahistine and selegiline.Our analysis showed that the primary endpoint (mean AUC of betahistine) increased in all three betahistine dosages:- 77-fold (95% CI: 42.3 to 141.5) by the combination of betahistine 24 mg plus selegiline 5 mg/d,- 86-fold (95% CI: 52.9 to 138.9) by the combination of 48 mg betahistine plus selegiline 5 mg/d, and- 108-fold (95% CI: 61 to 190.9) by the combination of 96 mg betahistine plus selegiline 5 mg/d compared to betahistine monotherapy .*ng/mL and for betahistine plus selegiline was 53.28 (+/-37.49) h*ng/ml. All the above results were statistically significant with a P < 0.0001, as shown in For instance, the mean (+/-SD) of the area under curve for betahistine 48 mg alone was 0.64 (+/-0.47) h*ng/ml are presented in The summary statistics are shown in When examining the effect of the combination treatment of betahistine and selegiline vs. betahistine alone on the half-life of betahistine, a one-way repeated measure ANOVA and the post hoc tests (Bonferroni) showed a statistically significant increase in the half-life of betahistine following the combination treatment of betahistine 24 mg and selegiline 5 mg/d. The half-life of betahistine was 1.9 times higher, 95% CI (1 to 3.5) after a combination treatment of selegiline 5 mg/d with betahistine 24 mg. In contrast, there was no statistically significant difference in the half-life of betahistine between the combination treatment and the single treatment of betahistine 48 & 96 mg on the plasma half-life, as shown in post hoc tests (Bonferroni) showed a statistically significant 50-fold increase in Cmax of betahistine following the combination treatment of betahistine 24 mg and selegiline 5 mg/d.When examining the effect of the combination treatment of selegiline and betahistine vs. betahistine alone on the Cmax of betahistine, a one-way repeated measure Anova and the There was no statistically significant difference between the combination treatment and the single treatment of betahistine 24, 48, and 96 mg on Tmax and elimination rate (ke) of betahistine.Based on the above reported findings, simulations of multiple doses with different dosages of betahistine/selegiline and dosing regiments were performed. In total, 14 adverse events (AE) were documented throughout the course of the trial . None ofIn total, 12 adverse events reported were classified as headache  and of these, 10 AEs were assessed as related to the combination treatment of betahistine and selegiline. One adverse event was classified as an upper respiratory infection  and assessed as not related to the trial treatment, and one adverse event was classified as leukopenia  and assessed as not related to the trial treatment. All adverse events that were assessed as related to the treatment were headaches. These headaches started 15 min to a few hours after the intake of betahistine after pre-treatment with selegiline and lasted a few hours without intake of concomitant medication. No AEs related to the therapy were observed due to therapy with betahistine without selegiline and due to concomitant treatment with 5 mg selegiline and 24 mg betahistine. Headaches related to the treatment were observed in six out of 15 subjects due to concomitant treatment with 5 mg selegiline and 48 mg betahistine and in six out of 15 subjects upon treatment with 5 mg selegiline and 96 mg betahistine.The major findings of this phase 1 pharmacokinetic study in 15 healthy volunteers are:Firstly, the MAO-B inhibitor selegiline  increases the bioavailability of orally administered betahistine  by a factor of between 77 and 108.Secondly, the half-life of around 30 min was largely unaffected; simulations showed that there was no accumulation over time with multiple doses.Thirdly, 14 AE were documented throughout the course of the trial. All of these were assessed as mild, and the outcomes of all AEs were classified as recovered or resolved. Twelve adverse events reported were classified as headache and, of these, 10 AEs were assessed as related to the combination treatment of betahistine and selegiline.Based on these findings and pharmacokinetic analysis, which showed an increase in the bioavailability of betahistine when combined with selegiline, as well as a favorable safety profile, phase II studies are needed and justified to evaluate the clinical efficacy. This combination can be useful for the preventive therapy of Menière's disease , acute tIn particular, for the preventive treatment of Menière's disease there is a high unmet medical need becausea) of its high prevalence: it is the second most frequent peripheral vestibular disorder  –18.b) Betahistine has already been used for decades in dosages of up to 48 mg tid, although it is not superior to placebo ; this fic) There is insufficient evidence for the effectiveness of many other measures that have been recommended, such as life-style changes, e.g., low salt diet , diuretiFinally, in terms of possible mechanisms of action of betahistine in Menière's disease, it was demonstrated in pre-clinical studies that it dose-dependently increases cochlear blood flow  which isCombining our pharmacokinetic data with published pharmacodynamic data enables us to refine the proposed mechanism of action of betahistine. Specifically, we can compare the Cmax values with the published affinities of the histamine receptors to evaluate target engagement. Based on radioactive binding experiments, H1, H3, and H2 receptors had Kd values for betahistine of 64 μM, 2 μM, and millimolar, respectively . In otheFinally, more broadly, these findings may also have an impact on the treatment of other diseases with betahistine. For instance, in high dosages it can improve cognition in schizophrenia .In the phase 1, single-center, open label study, only single dosages of betahistine and not the effects of repeated dosing was evaluated. However, the latter was studied by simulations. Only one dosage of selegiline was tested, which was half of the maximal dosage. As local authorities insist on ascending dosages for safety reasons, the study could not be randomized. For safety reasons, the sample size included only a small number of 15 healthy individuals.Future phase 2 trials need to pay attention to contraindication of selegiline, a MAO-inhibitor, which can lead to serotonergic effects and cannot be combined with tricyclics  or SSRI/SNRI anti-anxiety antidepressants. This could limit the usability in patients with MD and migraine as well as depression. Both medications are well tolerated, and the side effect profile is well-defined. Side effects of betahistine include headache and gastric complaints; selegiline can lead to dizziness, headache, bradycardia, and a mild elevation of liver enzymes, so monitoring is necessary.With regards to previous studies and analyses, which showed no effect of betahistine in MD in a dosage of up to 48 mg tid in comparison to placebo, and the uncertainties regarding the pathophysiology of Menière's disease and possible mode of action of betahistine in different diseases, it is uncertain if an increased bioavailability leads to a clinical effect. Therefore, placebo-controlled trials with a combination therapy in Menière's disease are needed. The same is true for acute vestibular syndrome or post-BPPV residual dizziness.This phase 1 trial shows that the MAO-B inhibitor selegiline increases betahistine bioavailability with no safety concerns. Whether the increased bioavailability has an impact on the preventive treatment of Menière's disease, acute vestibular syndrome, or residual dizziness after BPPV has to be evaluated in placebo-controlled trials.The original contributions presented in the study are included in the article/The studies involving humans were approved by Local Ethics Committees in Munich (19-992). The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.MS: Conceptualization, Data curation, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Writing—original draft. GC: Data curation, Formal analysis, Methodology, Validation, Visualization, Writing—original draft. IN: Investigation, Project administration, Resources, Writing—review and editing. UM: Supervision, Writing—review and editing. AA: Data curation, Formal analysis, Validation, Writing—review and editing. AG: Investigation, Writing—review and editing. NG: Investigation, Methodology, Project administration, Validation, Visualization, Writing—original draft.


In this study, grape pomace is used as an ingredient to fortify pasta. The grape pomace phenolic component is highly accessible and available for metabolization in the human gut. Hence, grape pomace can be exploited as a source of polyphenols and fiber for sustainable and dietary beneficial food production. Analyses of soluble and bound phenols and volatile compounds in raw and cooked pasta were performed. In the uncooked pasta fortified with pomace, the content of soluble and bound phenolic molecules increased significantly. During the cooking process, the bound phenols were lost, while the soluble phenols doubled. The whole grape pomace flour as a pasta ingredient increased the fiber component by at least double, increased the soluble polyphenol component by at least 10 times, and doubled the isoprenoids (toco-chromanols and carotenoids) while maintaining the unaltered fatty acid content after cooking. In accordance with the polyphenol content, antioxidant activity resulted higher than that of the control pasta. Analysis of volatile compounds in fortified pasta, both uncooked and cooked, indicated an improvement in aromatic profile when compared to the control pasta. Our results show that durum wheat pasta fortified with whole pomace flour has bioactive potential for the reuse of food industry byproducts. Pasta is among the most consumed foods worldwide, is part of the Mediterranean diet, and has a high level of acceptability regardless of the social class to which the consumer belongs. Italy is among the 10 countries with the highest consumption per capita .Due to its food importance, accessibility, and low value, in addition to being one of the most consumed products on a daily basis, pasta can be considered ideal for adding important ingredients to the diet. The search for alternative sources of healthier and more balanced food is increasing daily, leading the food industry to seek new technological processes that enable improved nutrition, and that do not have high processing costs, in order for them to reach all economic classes of consumers .In order to improve the nutritional value of foods, three approaches can be considered: (1) improving an existing formulation by adding and/or replacing certain ingredients; (2) developing new enriched products using raw materials previously selected as a source of essential nutrients; (3) evaluating the volatile fraction in final products. Each component to be added is characterized as an essential nutrient, providing healthiness to the consumer. In this way, the food can be classified as functional, which by definition is any ingredient in a food capable of providing health benefits, including the prevention and treatment of diseases .Therefore, the growing market for functional products, coupled with consumer interest in disease prevention, has driven the food industry to encourage direct research into the development and acceptability of new products in this regard. In this context, functional foods are gaining more and more success due to their possible health benefits .The food industry produces byproducts that may be considered an opportunity for sustainable valorization in order to minimize waste and ensure environmental defense . The reuMoringa oleifera leaf powder [About 50% of the GP is composed of grape peels, depending on the grape variety, as well as pedo-climatic and winemaking conditions ,12,14. Gf powder  added tof powder  suggestef powder . Furtherf powder . Anotherf powder .The addition of fiber-rich ingredients can have significant effects on dough rheological properties and on the final product texture, microstructure, and color. The effects of grape pomace on the composite flour and final product properties are relative to the addition level, while some negative effects of GP on dough rheology caused by gluten dilution can be minimized by particle size reduction ,24. FoodTrans-Resveratrol was obtained from ICN Biomedicals , whereas catechin, epicatechin, oenin, quercetin, quercetin-3-glucoside, and rutin were purchased from Extrasynthese . All other compounds such as solvents of analytical grade, were provided by Sigma-Aldrich .Vitis vinifera L. cv. Lambrusco (achieved after fermentation) and Fiano (without fermentation) were collected from a local commercial winery . Pomace samples were dried in an oven at 40 °C, for 48 h in the dark. The dried GP was milled by a laboratory sample mill  to obtain powder, and then passed through a 1 mm sieve.Wine pomaces (grape harvest year 2019) of www.pastificiodelduca.com, accessed on 31 May 2023, Parabita, Apulia Region, Italy). It was bronze-drawn pasta “fettuccine” made with durum wheat (Triticum durum L.) and water to help the development of gluten and salt. The development of pasta formulations consists of combining two ingredients: wheat flour and water (30%). This base formulation was modified by replacing 4% of wheat flour with 4% of GPF. For the preparation of pasta, the granulometry of wheat flour and mixed flours was adjusted to diameter ≤ 1 mm; then, the solid ingredients were mixed, before immediately adding the liquid ingredients until a homogeneous consistency of the dough was achieved. The drying process was carried out at temperatures below 40 °C for 36 h.Pasta (fortified and nonfortified samples) was produced by s local pasta factory (v/v ethanol. The combined supernatants (soluble phenolic fraction) were collected, evaporated, and hydrolyzed with 2 M NaOH for 4 h. The insoluble phenolic acids were extracted from pellets by hydrolysis with 2 M NaOH for 4 h. Samples were acidified to pH 2.0 with 12 M HCl and extracted twice with ethyl acetate. The upper phase was collected, evaporated, dissolved in 80% ethanol, and assayed by HPLC-DAD as reported by [Phenolic compounds (soluble or insoluble) were extracted from 1 g of raw and cooked pasta as described by . Brieflyorted by . Isoprenorted by . The finorted by .The Folin–Ciocâlteu method was carried out to evaluate the total amount of polyphenols. The total phenols (soluble or bound) in pasta extracts were assessed by determining the absorbance at 760 nm . Results•+-scavenging capacity. The analysis was performed as previously described by [The Trolox equivalent antioxidant capacity (TEAC) assay in extracts containing soluble or bound phenolic compounds was used to determine the total antioxidant capacity as the ABTSribed by . AbsorbaThe ORAC procedure was carried out following the procedure established by . The antg 10 min) and the organic phase was vaporized by a nitrogen stream. Lipids were subjected to fatty acid derivatization and analyzed by GC-MS as previous reported in [Total lipids were extracted from 0.1 g from pasta with 4 mL of n-hexane while stirring (3000 rpm) overnight at 4 °C. Samples were centrifugated . The CIELAB color space was used to determine the parameters L* from black (0) to white (100), a* (red (+a) to green (−a) color), and b* (yellow (+b) to blue (−b) color).Water absorption and cooking loss were determined as described by . Ten graCooking loss (CL) was assessed according to the AACC method 66–50 ; in an air oven at 105 °C ± 2 °C, the pasta cooking water was dried. The residue was weighed and expressed as a percentage of the starting material using the follow equation:The swelling index (SI) was determined as reported by  with somHeadspace solid-phase microextraction (HS-SPME) was employed to analyze volatile compounds according to  with somTo evaluate the effect of the pasta’s volatile compounds, the ratio between the single peak area and the total molecule area in a specific chromatogram, expressed as a percentage, was considered.Values represent the mean ± standard deviation of three independent replicates.t-test) was used to evaluate the statistical significance of the differences between the measured data by means of the SigmaStat software Version 3.1 . The principal component analysis of volatile compounds was carried out using the Statistica 6.0 software package.A parametric method , fibers (58.60 ± 1.70 g/100 g DW), and ash content (5.27 ± 0.32 g/100 g DW). The TP content was 4.97 ± 0.53 gGAE/100 g DW. The chemical composition of Lambrusco cv.-derived GPF was as follows: nitrogen substances (14.00 ± 1.05 g/100 g DW), fibers (53.00 ± 1.44 g/100 g DW), and ash content (23.34 ± 1.11 g/100 g DW). The TP content was 1.96 ± 0.02 gGAE/100 g DW. Fibers are the major component of grape pomace, as already stated by Antonić et al. [An HPLC analysis of soluble and bound phenols was performed A,B. In tIn accordance with the polyphenol amount, the addition of pomace flour increased antioxidant activity in comparison with the control pasta.The antioxidant power was evaluated in the phenol extracts (soluble and bound) of control pasta and pasta with pomace flour added, before and after cooking .In the raw control pasta, free phenols were scarce and almost undetectable, whereas, in the raw pasta with pomace flour, both white and red, they were definitely detectable. In the extracts of bound phenols, the TP, TEAC, and ORAC values were almost twice as high in the raw dough with added grape pomace flour as in the control.In each pasta sample, cooking greatly lowered the antioxidant capability. Moreover, bound phenols disappeared in all cooked pasta samples, and their antioxidant capacity could not be measured. After cooking, the antioxidant activity of soluble phenol fractions (both TEAC and ORAC) remained about 20 times higher than that of the control in pasta samples containing white pomace flour, and more than 10 times (from 11- to 13-fold) in pasta samples containing red pomace flour. The difference in antioxidant activity between the pasta sample obtained with white and red GP was related to the different phenolic compositions. Thus, it can be hypothesized that the different phenolic composition of the two pasta samples W and R led to a different response after cooking due to the presence of anthocyanins, which in the red, underwent degradation.The incorporation of GPF results in a compact microstructure of pasta ,12. ProtThe presence of whole GPF increased the fiber component before and after cooking . FurtherThe ash of food is the solid residue resulting from the complete combustion of organic matter. During combustion, sugars, starches, fibers, proteins, organic acids, vitamins, etc. are burnt, sparing the minerals, which accumulate in the ash in the form of oxides, carbonates, sulfates, and phosphates. Food ashes, if rich in oxides of potassium, magnesium, calcium, iron, etc., are defined as alkaline, as they can form alkaline aqueous solutions. The ash content reflects the mineral content shown in a previous paper . The ideThere was a significant increase in fiber content in the uncooked W and R pasta samples, as well as in the R samples after cooking, compared to cooked and uncooked control pasta samples. Cooking affects the fiber content of pasta, causing a slight loss of soluble dietary fibers ; howeverThe cooking time of our pasta (8–9 min) is not experimental. The data in Our starting material was total pomace flour  which involved lower work production and energy consumption with zero waste. The presence of the grape seeds enriched the flour with fatty acids and fat-soluble vitamins (vitamins A and E). Checking the shelf-life of the flour, we showed that the product was stable for 6 months at 4 °C . Table 5The addition of 4% GPF derived from both Fiano and Lambrusco cv. significantly modified the pasta color. As shown in SPME-GC/MS was applied to characterize the headspace volatile fraction of paste samples, enriched with pomace from two Italian varieties of Lambrusco (R) and Fiano (W), before and after cooking (1 and 2). As depicted in The significative ester percentage in cooked R was probably contributed by the pomace due to the typical fermentative activities of winemaking ,48. HighAmong the aldehydes and ketones, hexanal was found to be the most abundant compound in the volatile fraction of the pasta, as reported by others ,40, withPrincipal component analysis (PCA) was used as an exploratory statistical technique to investigate which variables play a major role in discriminating between cooked and uncooked samples. The control, on the other hand, both before and after cooking, appeared to be poorly correlated with the volatile molecules analyzed, suggesting a neutral sensory profile. PC2, on the other hand, highlighted the significant differences between the two samples fortified with red pomace, R1, and R2. R1, with a high PC2 (score 6.47) and negative PC1 (−2.18), correlated positively with esters; with cooking, esters decreased and aldehydes increased, characterizing sample R2, which clustered between positive PC1 and PC2.Multivariate analysis confirmed that most VOCs were associated with pasta enriched with aromatic Lambrusco grape pomace, followed by Fiano pomace.Vitis vinifera L.) in order to improve a zero-waste pathway for this wine-chain byproduct. Following this approach, GP can be considered a high-value resource to be converted into sustainable food and bio-functional ingredients, while fully respecting the circular economy concept.The results of this study indicate that is possible to obtain pasta with increased antioxidant activity, increased polyphenol content, increased tocochromanols and carotenoids, increased fiber content, and not modified or slightly increased fatty acid content, following the inclusion of grape pomace flour in the dough formulation. Moreover, pomace, red and/or white, can be successfully used as an additive in pasta samples, improving the volatile profile with increased esters, terpenes, alcohols, and aldehydes compared to the control. To our knowledge, this is the first paper on the production of bronze-drawn pasta made with controlled percentages of flour obtained from whole pomace flour including grape skins, grape seeds, and stems. In this study, we highlighted the integration of different approaches for valorizing the GP (


Objective: To compare the Duhamel and transanal endorectal pull-through (TERPT) procedures in the treatment of children with Hirschsprung’s disease. Methods: Studies comparing the Duhamel and TERPT procedures were included until 22 July 2023. R software (version 4.3.0) was used to perform the meta-analysis. Results: Ten studies with a sum of 496 patients were included. The length of postoperative hospital stay and incidence of postoperative constipation were longer and higher after the Duhamel procedure than the TERPT procedure . The incidence of postoperative anastomotic stricture was higher after the TERPT procedure than the Duhamel procedure (p = 0.0015). No significant differences were found in the incidence of postoperative fecal continence, fecal incontinence/soiling, anastomotic leak, or ileus between these two procedures. The operation time seemed to be similar for both procedures, but it became longer for the Duhamel procedure than the TERPT procedure after sensitivity analysis. While the incidence of postoperative enterocolitis seemed to be higher after the TERPT procedure, it became similar for both procedures in the subgroup analysis. Conclusions: The Duhamel procedure seems to be associated with a longer length of postoperative hospital stay, a higher incidence of postoperative constipation, and a lower incidence of postoperative anastomotic stricture than the TERPT procedure. However, the effect of these two procedures on the operation time and the incidence of postoperative enterocolitis remains unclear. With an incidence of 1 in 5000 live births, Hirschsprung’s disease (HSCR) is the most common gastrointestinal malformation causing intestinal obstruction in children ,3,4,5,6.The Duhamel procedure was first proposed in 1956 by Bernard Duhamel; it involves a longitudinal, side-to-side anastomosis of the posterior wall of the native aganglionic rectum with the anterior wall of the recruited ganglionic proximal colon and an end-to-side anastomosis of the posterior wall of the ganglionic colon with the retained rectal end . Then, IIn 1964, Soave  describeIn recent decades, the implementation of laparoscopy during the Duhamel and TERPT procedures has been reported to reduce trauma, loss of blood, intraoperative contamination, and intestinal adhesion ,30. HoweStudies were identified by searching databases, including PubMed (Medline), EMBASE, Cochrane Library, Web of Science, and EBSCO Host until 22 July 2023. Searches were conducted using Medical Subject Headings (MeSH) and free text words, including  AND . All articles were restricted to human studies written in English, and reference lists were searched for additional relevant articles. This systematic review was registered in PROSPERO with the registration number CRD42022357059.Inclusion criteria for the studies were set as follows: (1) studies comparing the functional outcomes after the Duhamel procedure or TERPT procedure, including randomized controlled studies (RCTs), retrospective studies, prospective studies, and observational studies; (2) a detailed description of the patients’ information, outcomes, and complications. The exclusion criteria were set as follows: (1) duplicate publications; (2) studies reporting findings on the same group of patients; (3) studies reporting fewer than 10 patients in any group; (4) studies only regarding total colonic aganglionosis or adults; (5) studies reporting reoperation patients; (6) single-arm studies; (7) studies without detailed raw data .The assessment of eligibility and risk of bias in included studies, as well as the raw data extraction, were performed by three independent reviewers . Discussion with other investigators (XB and FLW) resolved disagreements. The following data were collected for each suitable study: (1) general information: the first author, publication year, study location, study design, surgical technique, number of participants for each surgical procedure, participants’ gender and age; (2) baseline, benefits, and outcomes data: operation time, length of postoperative hospital stay, rate of postoperative fecal continence, fecal incontinence/soiling, constipation, enterocolitis, anastomotic stricture, leak, and postoperative ileus. Missing data were obtained by contacting the first and corresponding authors if possible.Two independent authors (LYY and LMQ) assessed the quality of the included studies using the Newcastle—Ottawa Scale (NOS) .I2 statistics. A common effect model  was used if I2 ≤ 50%, and a random effect model was used if I2 > 50%. Sensitivity analysis was conducted using a leave-one-out approach. Statistical significance was determined by a probability value of < 0.05.The protocol of this study was based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . The metThe search and screening process in this study is presented in n = 285 for the Duhamel procedure and n = 211 for the TERPT procedure) were included in the meta-analysis. Of these, 133 patients were from prospective studies [Finally, a total of 496 patients . The overall pooled analysis revealed that the operation time was similar between these two surgical procedures  (p < 0.0001) (Four studies (105 patients for the Duhamel procedure and 100 patients for the TERPT procedure) reported detailed data on the operation time and length of postoperative hospital stay ,19,20,22 0.0569) A, while  0.0001) B.2I did not change significantly (range: 93.3–98.9%), after omitting the data of operation time from Gunnarsdóttir et al. [p = 0.0311 and WMD = 95.22 min, 95% CI = 4.20 to 186.25, p = 0.0403, respectively). The detailed results are presented in the However, during the sensitivity analysis, we found that although the r et al.  and Sosnr et al.  . The overall pooled analysis revealed that the rates of postoperative fecal continence and fecal incontinence/soiling were similar between these two surgical procedures  (Five studies (167 patients for the Duhamel procedure and 111 patients for the TERPT procedure) reported detailed data on postoperative fecal continence ,19,23 T. The meactively) A,B.2I = 0%, p = 0.68). The overall pooled analysis revealed that the rate of postoperative constipation was higher in patients treated with the Duhamel procedure than in those treated with the TERPT procedure  (Six studies (172 patients for the Duhamel procedure and 99 patients for the TERPT procedure) reported detailed data on postoperative constipation ,20,21,24 0.0041) C.I2 = 40%, p = 0.14). The overall pooled analysis revealed that the rate of postoperative enterocolitis was lower in patients treated with the Duhamel procedure than in those treated with the TERPT procedure  (Six studies (161 patients for the Duhamel procedure and 151 patients for the TERPT procedure) reported detailed data on postoperative enterocolitis ,20,21,24 0.0033) A.I2 index decreased to 4% (p = 0.38), and the meta-analysis results indicated that the rate of postoperative enterocolitis was similar between these two surgical procedures   B. The deI2 = 0%, p = 0.99). The overall pooled analysis revealed that the rate of postoperative anastomotic stricture was lower in patients treated with the Duhamel procedure than in those treated with the TERPT procedure  (Five studies (123 patients for the Duhamel procedure and 107 patients for the TERPT procedure) reported detailed data on postoperative anastomotic stricture ,20,21,24 0.0015) A.I2 = 0%, p = 0.92 in postoperative anastomotic leak data and I2 = 0%, p = 0.97 in postoperative ileus data). The overall pooled analysis revealed that the rates of postoperative anastomotic leak and ileus were similar between the two surgical procedures  (Four studies (99 patients for the Duhamel procedure and 79 patients for the TERPT procedure) reported detailed data on postoperative anastomotic leak ,20,21,24ctively) B,C.A sensitivity analysis of the operation time and postoperative enterocolitis has been described above. In other sensitivity analyses, excluding each study in turn did not affect the meta-analysis results for the length of postoperative hospital stay or the rates of postoperative fecal continence, fecal incontinence/soiling, anastomotic stricture, constipation, anastomotic leak, and ileus.The Duhamel procedure and the TERPT procedure are commonly used for the treatment of HSCR ,11,12. TI2 = 98%, p < 0.01), we performed a sensitivity analysis and found that although the 2I did not change significantly, the meta-analysis results indicated that the operation time was longer for the Duhamel procedure than for the TERPT procedure after omitting the data from Gunnarsdóttir et al. [Initially, the results of the meta-analysis indicated that the operation time between the Duhamel and TERPT procedures was similar, which was consistent with prior systematic reviews ,37. Hower et al.  and Sosnr et al. . Notablyr et al.  and Sosnr et al.  was fewer et al. . Therefor et al. ,22 and tr et al.  study inAlthough the result of the comparison of the operation times was uncertain, the length of postoperative hospital stay seemed to be longer in patients treated with the Duhamel procedure, which was also consistent with previous studies ,37,38. IAs with several previous studies ,24,37,38Another reported postoperative complication that was similar to constipation was anastomotic stricture, which seemed to be higher in patients treated with the TERPT procedure. This may be related to the annular dissection and anastomosis in the anorectum of this procedure; other reported risk factors were anastomotic ischemia, anastomotic leakage, and cuff ischemia ,39,40,41One of the most serious postoperative complications was enterocolitis. The etiology of enterocolitis is unknown and is probably multifactorial . SeveralThis study has various limitations. First, most of the included studies were retrospectively designed with small sample sizes and different extent of disease, which could add bias to this meta-analysis. Second, the included studies spanned a wide timeframe (2004–2022). In the intervening years, the improvement of surgical techniques, perioperative management, complication prevention, and treatment would inevitably affect the overall prognosis of patients. This can also lead to bias and affect the analysis results. Third, the gap in age at operation among the included patients was also large (0.3–180 months), which may influence the incidence of complications (such as constipation and enterocolitis) and create bias. Fourth, variability in surgical techniques in the included studies, such as length of the rectal cuff, Swenson or Soave model for transanal procedure, and mechanical or manual coloanal anastomosis, may also create bias. Finally, the definition and evaluation criteria of complications were not standardized , which may also introduce bias and impact the results of the analysis.In conclusion, based on the data reported, the findings of this study indicated that the Duhamel procedure seems to be associated with a longer length of postoperative hospital stay and a higher incidence of postoperative constipation. The TERPT procedure seems to be associated with a higher incidence of postoperative anastomotic stricture. For the incidence of postoperative fecal continence, fecal incontinence/soiling, anastomotic leak, and ileus, these two procedures seem to be similar. However, based on the data analyzed, the differences in operation time and rate of postoperative enterocolitis between these two procedures were unclear. Further prospective studies with a larger sample size and adequate follow-up are needed to obtain more definitive conclusions.


Chronic lymphocytic leukemia (CLL) is one of the most common leukemias in adults. It has been associated with a number of dermatologic manifestations, such as leukemia cutis and erythema multiforme. Among the rarer of these findings is eosinophilic dermatosis of hematologic malignancy (EDHM). EDHM was originally characterized as a hypersensitive insect bite-like reaction, despite most patients having no distinct recollection of being bit or having any risk of exposure. Typically, EDHM presents as a pruritic, erythematous eruption, often with papulovesicular lesions, throughout the body. Due to its relapsing course, a number of treatment methods have been proposed, but no standard of care has been established. In this report, we present a recalcitrant case of EDHM in a patient with CLL that responded well to treatment with narrow-band ultraviolet B (NBUVB) light therapy. Chronic lymphocytic leukemia (CLL), an adult leukemia caused by malignant proliferation of B-cells, is associated with a number of dermatological conditions, such as leukemia cutis and erythema multiforme ,2. DermaThis case report was previously presented at Texas A&M University Student Research Week on 03/24/22, Texas Dermatological Society Annual Spring Meeting on 04/29/22, Baylor Scott & White Medical Center - Temple Research Scholars Day on 05/05/22, Brigham and Women’s Hospital Dermatology Medical Student Virtual Symposium on 05/25/22. The patient provided verbal and written consent for treatment and for the publication of the report with photos.A 72-year-old female patient with a history of CLL presented with a several-week history of large, erythematous, pruritic wheals on the extremities that had appeared after she worked in the garden. At this time, although the patient had no distinct recollection of insect bites, she was diagnosed with exuberant insect bite reactions based on the belief that patients with CLL may develop significant local reactions to insect bites. The lesions resolved within a couple of months of topical steroid use as needed up to twice a day.Two years later, she returned to the clinic with a similar but more severe presentation of extremely pruritic, burning pink papules, wheals, and small bullae with subsequent erosions located on the extremities, palms, chin, cheeks, and forehead Figures , 2. She A biopsy of the left leg was performed and pathology results showed a small-vessel vasculitis with eosinophils, papillary dermal edema, and intraepidermal vesicles. CBC and CMP were within normal limits. Direct immunofluorescence (DIF) was not performed, but serum bullous pemphigoid antigens BP180 and BP230 were negative. Based on clinical presentation, concurrent diagnosis of CLL, and findings on the biopsy specimen, a diagnosis of EDHM was made. Therapy was initiated with topical steroids followed by oral steroids, oral doxycycline 100 mg twice daily, and nicotinamide 500 mg twice daily, as these have all been suggested to have therapeutic benefits in previous reports of EDHM ,10-12. HNBUVB has been described in use for the treatment of EDHM  and the Although EDHM is a rare entity, it may cause significant distress and detriment to patients and negatively affect their quality of life. An existing hypothesis for the pathophysiology of EDHM describes the interplay of leukemic cells with interleukin-4 (IL-4) and interleukin-5 (IL-5). These promote lymphocyte proliferation and tissue eosinophilia, which then drive the development of the lesions -16. AnotEDHM most commonly occurs in patients between the ages of 40 and 60 years, and it may occur before, during, or after the diagnosis of CLL, lasting for a variable duration of time ,11,12. DOn pathology, EDHM demonstrates tissue eosinophilia with dermal perivascular lymphocytic infiltrate with possible spongiosis and intraepidermal vesicles that contribute to the blister-like appearance of the lesions ,4,11,15.Largely due to its refractory, relapsing nature, no standard treatment regimen has been described for EDHM. Suggested treatment options include topical and systemic corticosteroids, subcutaneous dupilumab, monoclonal antibodies, IVIG, oral antibiotics, antihistamines, and phototherapy -14. WhilLimitations of this case and our hypothesis that NBUVB may be considered a safe and effective treatment option for EDHM include concurrent use of IVIG, and the possibility of spontaneous resolution . Our patient started IVIG not long after the onset of lesions and reported only mild improvement with this treatment (which was prescribed for her CLL). Once she started NBUVB, she saw rapid improvement in current lesions and a decrease in the development of new lesions. Additionally, when her EDHM relapsed several months later, she restarted NBUVB therapy and experienced a similar positive response. This reproducibility further validates the utility of this treatment regimen.The patient described here initially presented with insect bite-like lesions and a history that corroborated a diagnosis of exuberant reaction to insect bites. She presented two years later with biopsy-proven EDHM. Treatment was attempted with topical and systemic steroids, oral antibiotics, and oral nicotinamide, with responses ranging from poor tolerance to transient improvement, at best. While she was undergoing concurrent therapy for CLL, it was only after the initiation of NBUVB that the patient experienced significant improvement in her existing lesions, with lasting resolution after the successful treatment of a second episode. Based on our experience, NBUVB may be considered as an efficacious first-line treatment for EDHM.