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Introduction During the COVID-19 pandemic, health workers represented a group particularly vulnerable to work-related stress, but prevention and management of psychiatric symptoms are still under evaluation. Neurofeedback is a safe and non-invasive neuromodulation technique with the target of training participants in the self-regulation of neural substrates underlying specific psychiatric disorders. Protocols based on the increase of alpha frequencies, associated with the process of relaxation, are used for the treatment of stress, anxiety and sleep disturbances.ObjectivesThe aim of the present study was to assess the effectiveness of an alpha-increase NF protocol for the treatment of stress in healthcare workers exposed to the COVID-19 pandemic.MethodsEighteen medical doctors belonging to the Sacco Hospital were recruited during the COVID-19 health emergency and underwent a 10 sessions NF alpha-increase protocol during two consecutive weeks. The level of stress was assessed at the beginning (T0) and at the end (T1) of the protocol through the following questionnaires: Severity of Acute Symptoms Stress (SASS), Copenhagen Burnout Inventory (CBI), Pittsburgh Sleep Quality Index (PSQI), Brief-COPE. Statistical analyses were performed with Paired Samples t-Test for continuous variables, setting significance at p < 0.05.ResultsA significant increase in alpha waves mean values between T0 and T1 was observed. In addition, a significant reduction in the PSQI test score between T0 and T1 was observed.ConclusionsAlpha-increase protocol showed promising results in terms of stress modulation, sleep quality improvement and safety profile in a pilot sample of health-care workers. Larger controlled studies are warranted to confirm present results.

Summary Chromosomal anomalies, like Robertsonian and reciprocal translocations, represent a big problem in cattle breeding as their presence induces, in the carrier subjects, a well-documented fertility reduction. In cattle, reciprocal translocations (RCPs, a chromosome abnormality caused by an exchange of material between non-homologous chromosomes) are considered rare as to date only 19 reciprocal translocations have been described. In cattle, it is common knowledge that the Robertsonian translocations represent the most common cytogenetic anomalies, and this is probably due to the existence of the endemic 1;29 Robertsonian translocation. However, these considerations are based on data obtained using techniques that are unable to identify all reciprocal translocations, and thus, their frequency is clearly underestimated. The purpose of this work is to provide a first realistic estimate of the impact of RCPs in the cattle population studied, trying to eliminate the factors that have caused an underestimation of their frequency so far. We performed this work using a mathematical as well as a simulation approach and, as biological data, we considered the cytogenetic results obtained in the last 15 years. The results obtained show that only 16% of reciprocal translocations can be detected using simple Giemsa techniques, and consequently, they could be present in no

The employment ofCamellia sinensis L. whole plant extract as food supplement in livestock nutrition has been suggested in order to prevent usual livestock intestinal diseases. The aim of the present research was to test the effects of such plant extract on the composition of pig faecal microbiota. Preliminaryin vitro fermentation trials evidenced in mixed pig faecal cultures, supplemented with the tested extract, an increase of total anaerobe (p=0.02) and aerobe (p=0.03) bacteria, and a decrease of clostridia (p=0.04) compared to control cultures. Afterwards we investigatedin vivo the effects on piglet faecal microbiota of a diet added with 250 mg/kg ofCamellia sinensis whole plant extract. A control diet without the plant extract, but added with antibiotic (sulphadiazine, trimethoprim, and tiamulin) was used for the comparison. Microbiological analyses of faecal samples collected after 60 days of the experimental feeding, evidenced a decrease of clostridia (p=0.001) and enterococci (p=0.04) counts in the faeces of animals fed with the experimental diet, compared to those fed with the control diet. These results show that theCamellia sinensis L. whole plant extract is able to reduce the number of some potential pathogenic bacteria in piglet gut and hence might improve animal health.

Clinical animal cytogenetics development began in the 1960’s, almost at the same time as human cytogenetics. However, the development of the two disciplines has been very different during the last four decades. Clinical animal cytogenetics reached its ‘Golden Age’ at the end of the 1980’s. The majority of the laboratories, as well as the main screening programs in farm animal species, presented in this review, were implemented during that period, under the guidance of some historical leaders, the first of whom was Ingemar Gustavsson. Over the past 40 years, hundreds of scientific publications reporting original chromosomal abnormalities generally associated with clinical disorders (mainly fertility impairment) have been published. Since the 1980’s, the number of scientists involved in clinical animal cytogenetics has drastically decreased for different reasons and the activities in that field are now concentrated in only a few laboratories (10 to 15, mainly in Europe), some of which have become highly specialized. Currently between 8,000 and 10,000 chromosomal analyses are carried out each year worldwide, mainly in cattle, pigs, and horses. About half of these analyses are performed in one French laboratory. Accurate estimates of the prevalence of chromosomal abnormalities in some populations are now available. For instance, one phenotypically normal pig in 200 controlled in France carries a structural chromosomal rearrangement. The frequency of the widespread 1;29 Robertsonian translocation in cattle has greatly decreased in most countries, but remains rather high in certain breeds (up to 20–25% in large beef cattle populations, even higher in some local breeds). The continuation, and in some instances the development of the chromosomal screening programs in farm animal populations allowed the implementation of new and original scientific projects, aimed at exploring some basic questions in the fields of chromosome and/or cell biology, thanks to easier access to interesting biological materials (germ cells, gametes, embryos ...).

TLQP-21 is a neuropeptide expressed in the brain that is involved in the control of energy homeostasis. In preliminary experiments we have observed that TLQP-21 can modulate macrophage function. In Acute Respiratory Distress Syndrome (ARDS) macrophage seems to play a critical role, contributing to lung remodeling.

Adjuvant trastuzumab (AT) dramatically improved HER2-positive breast cancer prognosis. Relapsed disease after AT has different patterns and information is available from observational studies. In this Review Chemotherapy regimens combined to anti-HER2 blockade are discussed, focusing in particular the role of anthracyclines, taxanes and capecitabine. The use of trastuzumab beyond progression and the role of other anti-HER2 agents like lapatinib, pertuzumab and T-DM1 are explored, as also dual blockade and in trastuzumab resistant Patients. Metastatic "de novo" HER2 Luminal (co-expression of HER2 and hormone receptors) Patients are eligible for anastrozole and trastuzumab but if pretreated with trastuzumab they are also eligible for lapatinib and letrozole. In any case endocrine treatment plays a complementary role to chemotherapy which remains pivotal. The last topic explored is treatment options for patients with brain metastases where both trastuzumab given concurrent with radiotherapy or lapatinib and capecitabine appear as potentially active.

Ichthyosis is a heterogeneous group of keratinization disorders reported both in human and animals. Two rare, inherited forms have been reported in cattle, both characterized by autosomal recessive transmission. Because mutations of transglutaminase 1 (TGM1) gene are associated with autosomal recessive ichthyosis in people, this gene was investigated as a candidate for the diseases in cattle. Three different polymorphisms were identified in 5' end region of cattle TGM1. Marker homozygosity was not found among affected calves. Linkage analysis excluded (logarithmic odds [LOD] score -2.0) TGM1 as the cause for ichthyosis phenotype in the analyzed Chianina cases.

Although the majority of patients with anorexia nervosa and bulimia nervosa develop these disorders in their teens and 20s, some patients develop an eating disorder in their 30s, 40s, or 50s. We present a subgroup of patients with the following pattern of symptoms and historical detail: (1) severe sexual and physical abuse by family members; (2) relatively good premorbid professional and marital adjustment (considering later difficulties) though characterized by (3) hypomania, binge eating, and morbid obesity. A pronounced shift in eating behaviors follows (4) medical trauma (e.g., injury, cancer, surgery) that occurs after age 30, interrupts previous hypomanic adaptation, and leads to severe restriction, purging, and dramatic weight loss (e.g., 100 lb). Although only one patient met full criteria for anorexia nervosa, weight loss and starvation were serious enough to provoke further medical crises in all patients. Finally, (5) during both weight loss and weight restoration patients demonstrated significant dissociative disturbance, including dissociated mood and personality states (i.e., multiple personality disorder), self-destructive behavioral episodes repeating early trauma, and avoidance of food as a way to manage PTSD symptoms.

Robertsonian translocation (rob) involving chromosomes 1 and 29 represents the most frequent chromosome abnormality observed in cattle breeds intended for meat production. The negative effects of this anomaly on fertility are widely demonstrated, and in many countries, screening programs are being carried out to eliminate carriers from reproduction. Although rob(1;29) was first observed in 1964, the genomic structure of this anomaly is partially unclear. In this work, we demonstrate that, during the fusion process, around 5.4 Mb of the pericentromeric region of BTA29 moves to the q arm, close to the centromere, of rob(1;29). We also clearly show that this fragment is inverted. We find that no deletion/duplication involving sequences reported in the BosTau6 genome assembly occurred during the fusion process which originates this translocation. Acknowledgements. This study was partially supported by the Rural Development Plan under the Project PSR, Misura 214, e2 of Campania, "Razze Autoctone a Rischio di Estinzione della Regione Campania-- RARECa"

Cytogenetic analysis of a phenotypically normal young bull from the Marchigiana breed revealed the presence of an abnormal chromosome. The finding of one oversize chromosome in all metaphases, associated with a 2n = 60, XY karyotype, suggested that a reciprocal translocation had occurred. RBG-banding and FISH analyses, using specific bovine BAC probes, identified a de novo reciprocal translocation t(4;7)(q14;q28). The presence of rcp(4;7) was confirmed by FISH experiments using BTA4 and BTA7 whole chromosome probes. An array-CGH analysis (Agilent 244A) using a bovine custom design was performed to investigate if the translocation was associated with loss or gain of genetic material. The absence of a concomitant deletion or duplication at the break points allowed the balanced state of the translocation to establish. The analysis also revealed the presence of several CNVs throughout the genome. To our knowledge this is the first time the balanced condition of a cattle RCP has been ascertained using the array-CGH approach.

Routine cytogenetic investigations of the Chianina cattle (BTA) breed revealed the presence of longer and smaller chromosomes than the largest (BTA1) and smallest (BTA29) chromosomes in the cells of a young, normal-looking bull used for reproduction. Application of both RBA-banding and Ag-NOR techniques, as well as the use of the FISH technique and specific molecular markers of both BTA11 (IL1B, ASS and LGB) and BTA21 (SERPINA and D21S45) established that these two abnormal chromosomes were the product of a reciprocal translocation between BTA11 and BTA21. Both der(11) and der(21) were C-band positive and the chromosome regions affected were rcp(11;21)(q28;q12). The young bull had a normal body conformation, including external genitalia, normal levels of testosterone (as in the control) and non-detectable levels of both 17 beta-estradiol and progesterone (as in the control). The animal never showed libido in the presence of both males and females in oestrus. After slaughter at 18 months, histological evaluation revealed normal organized testes, seminiferous tubules and epididymis but with poor proliferative germ cells consisting mainly of spermatogonia, middle pachytene spermatocytes and early spermatids with late spermatids and spermatozoa being very rare.

The oral mucosae of the lips and muzzle appeared histologically normal. Mitotic chromosomes were visualised with conventional staining and RBA-banding from non-synchronised cultures of peripheral blood lymphocytes of the affected calves and their relatives (De Grouchy and others 1964, Dutrillaux and others 1973). No abnormality was detected in either the structure or the number of chromosomes. Genealogical data from the Chianina Breed Stud Book, from the Associazione Nazionale Allevatori Bovini Italiani da Carne (ANABIC), were collected to analyse the pedigrees of the two affected animals and of nine other newborn Chianina calves showing the same condition (Fig 3). Wherever possible, blood samples were collected from the affected animals and their sires, dams and siblings. Parentage relationships were verified by microsatellite analysis with StockMarks for Cattle Paternity PCR typing kits (Applied Biosystems). Forty-nine animals of the familial group were evaluated. Analysis of the pedigree information revealed that all clinical cases were generated by consanguineous matings. In particular, three bulls were acknowledged as being disease carriers: bulls 23, 30 and 34 (Fig 3). Bull 23 sired three affected male calves (26, 28 and 48) and two healthy female calves (31 and 35); bull 30 sired two affected male calves (27 and 49) and two healthy calves, one male (34) and one female (46); and bull 34 sired six affected calves, four males (38, 40, 41 and 47) and two females (43 and 44) and four healthy calves, three males (37, 42 and 45) and one female (39). None of the carIchthyosis in Chianina cattle

Sixteen carriers of rob(1;29) (one of which was homozygous) from six different breeds (four Italian and two Portuguese), two heterozygous carriers of rob(26;29), three river buffaloes and two sheep were cytogenetically investigated in this study by using banding and FISH-mapping techniques (the latter only in cattle and river buffalo). Single- and dual- colour FISH were used with bovine probes containing both INRA143 (mapping proximally to BTA29) and bovine satellite (SAT) DNA SAT I, SAT III and SAT IV (mapping at the centromeric regions of cattle chromosomes). The combined use of these probes, the comparison of rob(1;29) with the dicentric rob(26;29) and with both river buffalo and sheep chromosomes (biarmed pairs) allowed us to hypothezise that rob(1;29) originated from complex chromosomal rearrangements through at least three sequential events: (a) centric fusion with the formation of a dicentric chromosome; (b) formation of a monocentric chromosome with loss of SAT I from both BTA1 and BTA29, most of SAT IV from BTA29 and, probably, some repeats of SAT III from BTA1; (c) double pericentric inversion or, more probably, a chromosome transposition of a small chromosome segment containing INRA143 from proximal p-arms to proximal q-arm of the translocated chromosome.

During normal cytogenetic investigations on the Chianina cattle (BTA) breed, a normal looking young bull was found to carry an abnormal Y chromosome which was a product of a reciprocal translocation between chromosomes Y and 9. This was revealed by both CBA- and RBG-banding techniques and was clearly confirmed by FISH-mapping analysis with IDVGA50 (which paints the complete Yq arm in a normal Y), as well as with AMD1, CGA, IGF2R (mapping to BTA9q16, BTA9q22 and BTA9q27-->q28, respectively) and SRY (mapping to normal BTAYq23). Analysis on sperm from four different samples revealed azoospermia in the carrier, indicating that the rcp(Y;9) induces sterility in the bull. Copyright 2002 S. Karger AG, Basel

This report concerns the cytogenetic analysis, using both C-banding and fluorescence in situ hybridisation techniques, of a sterile mare. Results obtained revealed a 2n = 65, XXX condition with no sign of mosaicism. The work supports the suggestion that X trisomy, rare in horse, causes infertility in mares and is not associated to other clearly visible phenotypic features.

The present paper describes two cases of an XXY condition in Chianina cattle. Both young bulls were routinely investigated cytogenetically before entering progeny test stations. Every cell examined in the blood cell cultures showed an XXY constitution. The histological study of the gonads, performed on only one bull, showed degradation of the seminiferous tubules. Only Sertoli cells and hyperplastic interstitial cells were observed.

A new Robertsonian translocation has been found in cattle. A bull from Marchigiana breed (central Italy) was found to be a heterozygous carrier of a centric fusion translocation involving cattle chromosomes 13 and 19 according to RBA-banding and cattle standard nomenclatures. CBC-banding revealed the dicentric nature of this new translocation, underlining the recent origin of this fusion. In fact, both the bull's parents and relatives had normal karyotypes. In vitro fertilization tests were also performed in the bull carrying the new translocation, in two bulls with normal karyotypes (control) and in four other bulls carrying four different translocations.

Summary In pig, the frequency of intersexuality ranges from 0.1 to O.6%, depending on the breed. In a closed pig herd at INRA an intersex condition was observed in 0.75% of ‘females’. The present study describes 11 animals with a 38XX karyotype and the presence of testicular tissue. Phenotypically, all presented with abnormal external or/and internal genitalia. Southern blot analysis with Y-specific probes (SRY and ZFY) revealed the absence of Y material in all animals tested. By polymerase chain reaction (PCR) amplification, 10 of 11 intersex pigs lacked the SRY gene in gonad DNA. These data are compatible with an autosomally (or pseudoautosomally) determined mechanism. Moreover, analysis of familial cases seemed to indicate that 38XX male pseudohermaprodites and 38XX true hermaphrodites may represent alternative manifestations of the same genetic defect.

The fumonisins, produced by fungi that infest foodstuffs, in particular corn, are responsible for a series of illnesses and toxicosis in various species of animals, including humans. There is still not detailed information about the genotoxic and mutagenic activity of Fumonisin B1 (FB1), but it is clear that it interferes with growth control, differentiation and cellu- lar apoptosis. The purpose of this study was to assess the genotoxic potential of Fumonisin B1 using in vitrocultures of bovine lymphocytes, through the calculation of the ‘mitotic index’ (MI), the frequency of ‘sister chromatid exchange’ (SCE) and the ‘micronucleus test’ (MN). The bovine lymphocytes were exposed to different concentrations of FB1 (25, 50 and 100 µM) in order to find out which amount is sufficient to cause a reduction in the mitotic potential of the cells, the onset of MN and a higher frequency of SCE. The results obtained show a considerable reduction in the ‘mitotic index’ with a FB1 concentration of 50 µM, an increase in the frequency of MN with a concentration of 50 µM and a significant increase in the SCE with a concentration of 100 µM. In the light of the information we have obtained, compared with that of other Authors, we feel that the genotoxic poten- tial of FB1 has been underestimated until now and should, therefore, be reconsidered.

Inherited Ichthyosis, Chianina, Causative mutation, Genetic screening.Inherited Ichthyosis is a genetic disorder reported in both humans and animals, including bovines. Two inherited forms were reported in cattle and both are transmitted in an autosomal recessive manner: Ichthyosis Fetalis (IF) and Ichthyosis Congenita (IC). A causative mutation of IF in Chianina cattle was recently indentified in the ABC12 gene. This work reports the first genetic screening using this recently available genetic test on Chianina cattle. Tests were performed on both the population of farm breeding selected young bulls (131 samples randomly chosen) and high breeding value sires (16 samples). Results confirm a low total prevalence of carriers in the selected sire population (2/131; 1.5%) and the presence of the disease allele among the high value selected sires (1/16; 6.3%). This result strengthens the importance to continue the genetic screening program, particularly in performance tested bulls approved for use in AI or natural service.

Five cattle, two of the Podolian breed, two Grey Alpine, and one Chianina,all of which were heterozygous carriers of three centric fusion translocations(rob-l;29, rob-4;8, rob-25;27), underwent cytogenetic investigation. The use of G-, R- and C-banding patterns, combined with the FISH technique by using both type I molecular markers and some human chromosome painting probes, allowed the precise identification of chromosomes involved in the chromosomal abnormalities,compared to previous reports. While the chromosomes involved in the well known rob (1;29) were confirmed, BTA6 (not BTA4) was involved in the second translocation. Furthermore, BTA26 and BTA29 (not BTA25 and BTA27) were involved in the third translocation. C-banding patterns confirmed the mono-centric nature of rob (1;29) and revealed the dicentric nature of both rob(6;8) and rob(26;29). The importance of these marker chromosomes in bovid chromosome nomenclatures is also discussed.