Effects of different hormonal treatments on spermatogenesis advancement in hatchery-produced greater amberjack Seriola dumerili (Risso 1810).

In earlier studies, wild-caught greater amberjack Seriola dumerili (Risso, 1810) males reared in sea cages showed gametogenesis impairment and low sperm production and quality. Here, we (a) examined if F1 hatchery-produced males reared in sea cages also exhibit reproductive dysfunctions and (b) evaluated the effects of gonadotropin releasing hormone agonist (GnRHa) administration through injections (GnRHainj) or sustained-release implants (GnRHaimpl), and human chorionic gonadotropin (hGC) injections on spermatogenesis/spermiation enhancement. Fish were given a hormone treatment just prior to the spawning season, and were transferred to land-based tanks, according to an established spawning induction protocol. Blood samples (n = 6) were obtained on Days 0, 7 and 13 after treatment. Testis samples were obtained on Days 0 (n = 4) and 13 (n = 2 per treatment). The fish prior to their transfer from the sea cages to the land-based tanks, exhibited a low gonadosomatic index, altered sex steroid hormone profile and high density of testicular apoptotic cells. After transfer to tanks, there was a general depression of sex steroid plasma levels parallel to an increase in cortisol concentrations. Despite the negative effect on steroidogenesis by the transfer from the sea, the hormonal treatments increased the number of fish from where sperm could be obtained, as well as testis growth, and reduced testicular apoptosis. Treatment with hCG resulted in the most significant changes in spermatogenesis, while GnRHaimpl appeared to induce less intense, but likely longer-lasting effects. The study indicated that F1 hatchery-produced males also exhibited reproductive dysfunctions as wild-caught captive-reared greater amberjack, and that the observed positive effects of the hormone treatments on spermiation/spermatogenesis were likely mediated by factors other than sex steroid hormones.

Liver melanomacrophage centres as indicators of Atlantic bluefin tuna, Thunnus thynnus L. well-being.

Melanomacrophage centres (MMCs), located in different organs of non-mammalian vertebrates, play a role in the destruction, detoxification or recycling of endogenous and exogenous materials. Cytochrome P450 monoxygenase 1A (CYP1A) is involved in xenobiotics biotransformation, and its liver expression is considered as a biomarker for detecting exposure to environmental pollutants. Atlantic bluefin tuna (ABFT), Thunnus thynnus L., liver samples were collected from: wild animals caught in the eastern Atlantic; juveniles reared in the central Adriatic; juveniles reared in the northern Adriatic; adults reared in the western Mediterranean. The samples were processed for basic histology, histochemistry and for CYP1A immunodetection. An unexpected high density of MMCs, containing ferric iron and lipofuscin-ceroids, was detected in the juveniles sampled in the northern Adriatic Sea. These individuals showed also a strong anti-CYP1A immunopositivity in hepatocytes and in the epithelium of bile ducts. This study supports the utility of MMCs as biomarkers of fish 'health status' and gives concern for a potential contaminant accumulation in ABFT.

In vitro effect of isotocin on ovarian tunica albuginea contractility of gilthead seabream (Sparus aurata L.) in different reproductive conditions.

Contractions of ovarian tunica albuginea, the teleostean cystovary wall layer containing smooth muscle fibres, facilitate oocytes and fluids movements within the ovary, oocytes ovulation and spawning. Fish isotocin, the homologue hormone of mammalian oxytocin, plays a significant role in ovulation, oviduct contraction and spawning. In the present study, ovarian wall spontaneous contraction, as well as isotocin in vitro effect on tunica albuginea contractility, was analysed in female seabream in different reproductive conditions: vitellogenesis, regressing (post-spawning) and extensive atresia. Tunica albuginea spontaneous contractility was recorded using ovary wall strips mounted in an organ bath containing modified Ringer's solution. The strips were then exposed to cumulative doses of isotocin (6, 30, 60 µg/ml). Female seabream in regressing condition exhibited the highest level of tunica albuginea spontaneous contraction amplitude compared with the other two groups. Only fish in vitellogenesis state showed a significant increase in contraction amplitude after isotocin administration at the dose of 30 µg/ml. The same group exhibited also a significant isotocin dose-dependent decrease in the contractile frequency. These results confirm the involvement of isotocin in stimulating tunica albuginea contractile activity during the oestrogen-regulated phase of vitellogenesis, whereas the absence of significant effects of isotocin on ovarian contractility in fish at the regressing state might be ascribed to the occurrence of a contractile activity autonomously regulated by the internal pacemaker system. The absence of exposed isotocin receptors could explain the lack of effects of the isotocin administration in seabream showed extensive atresia of the follicular cells.

Ovarian contractility in reared gilthead seabream (Sparus aurata L.) in different phases of the reproductive cycle.

Spontaneous ovarian tunica albuginea contractility was evaluated in gilthead seabream (Sparus aurata L.) at different phases of the reproductive cycle. Fourteen adult females were sampled from February to November 2012 in a commercial fish farm, and ovaries were removed and processed for histological and contractility analyses. Fish reproductive stages were evaluated on haematoxylin-eosin-stained ovary sections or by simple macroscopic observation of hydrated oocytes in spawning individuals. Tunica albuginea spontaneous contractility was recorded by using ovary wall strips mounted in an organ bath containing modified Ringer's solution. Ovary macro- and microscopic analyses allowed the identification of three different reproductive conditions: vitellogenesis, spawning and regressing. The gilthead seabream tunica albuginea was capable to contract spontaneously, and significant differences were found in mean contraction amplitude among the three reproductive states, with the highest value recorded in individuals in regressing condition and the lowest in individuals at spawning stage. No differences in mean contractility frequency among the three different groups were found. Possible involvement of spontaneous contractility in facilitating developing follicle advancement towards the ovarian lumen within the ovary and in supporting recovery of regressing ovaries may be hypothesized. The low contractility observed during the final oocyte maturation and spawning phases does not seem to support a role of tunica albuginea during ovulation, which could conversely involve theca cell contraction. Alternatively, possible single instantaneous contractions of tunica albuginea muscle fibres, not detected in the present study, could occur during ovulation in response to neuro-hormonal stimulations; a role of abdominal wall musculature in ovary "squeezing" and consequent release of ovulated eggs cannot be excluded.

The experience of the rigid lockdown in the dental emergency room and urgency care during COVID-19 pandemic: a transnational multicenter observational study.

OBJECTIVE: The COVID-19 pandemic had a major impact on our lives all over the world. Changes have occurred in daily life as well as in all medical services. The aim of the present study was to evaluate the emergency accesses in four universities' emergency services during the lockdown period from March to June 2020 during the COVID-19 pandemic. SUBJECTS AND METHODS: A cross-sectional study was carried out on 44,787 patients to evaluate the emergency services of university centers. The medical data of Medical Emergency Service Data (MESD) were assessed by five independent operators considering the epidemiological findings for statistics methods. RESULTS: A lower level of emergency access was reported in March-July compared to the pre-COVID period. The epidemiological data confirmed that female pathologies were more frequent compared to male patients. A fluctuation for almost all urgent healthcare centers was detected, showing one/two peaks per year during the years 2017-2019. The COVID-19 pandemic period did not influence the variety of pathology detected. CONCLUSIONS: After the lockdown period, the emergency services slowly increased in cases. The pre-COVID period showed an overlapping of the most frequent pathologies compared to the post-COVID period: periodontitis (Bari and Tirana), dental fractures (Bari and Bucharest), odontogenic abscess (Bari, Cluj and Tirana).

The human microbiota key role in the bone metabolism activity.

Data collection has suggested a complex correlation between the gut microbiota (GM) and bone homeostasis involving host-microbiota crosstalk. Although the GM is known to affect bone metabolism, the mechanisms linked with these effects remain unclear. The aim of this review is to current insight advances regarding how gut-derived hormones regulate bone homeostasis in humans, emphasizing gut-bone axis and bone regeneration. The GM may be engaged in bone metabolism and fracture risk. Additional investigations of the fundamental microbiota-related pathways in bone metabolism may uncover treatment strategies and enable the prevention of osteoporosis. A better knowledge of gut hormones' action on bone homeostasis may lead to new strategies for preventing and treating skeletal frailty related to age.

GnRHa-mediated stimulation of the reproductive endocrine axis in captive Atlantic bluefin tuna, Thunnus thynnus.

A controlled-release implant loaded with GnRH agonist (GnRHa) was used to induce spawning in Atlantic bluefin tuna (Thunnus thynnus) during two consecutive reproductive seasons. The fish were implanted underwater and sampled between days 2 and 8 after treatment. At the time of GnRHa treatment, females were in full vitellogenesis and males in spermiation. There was a rapid burst of pituitary luteinizing hormone (LH) release at day 2 after treatment in GnRHa-treated fish, and circulating LH remained elevated up to day 8 after treatment. In contrast, control fish had significantly lower levels in the plasma, but higher LH content in the pituitary, as observed in many other cultured fishes that fail to undergo oocyte maturation, ovulation and spawning unless induced by an exogenous GnRHa. Plasma testosterone (T) and 17ß-estradiol (E(2)) were elevated in response to the GnRHa treatment in females, while 11-ketotestosterone (11-KT) but not T was elevated in males. Even though oocyte maturation and ovulation did occur in GnRHa-induced fish, no significant elevations in 17,20ß-dihydroxy-4-pregnen-3-one (17,20ß-P) or 17,20ß,21-trihydroxy-4-pregnen-3-one (20ß-S), in either the free, conjugated or 5ß-reduced,3α-hydroxylated forms was observed in fish sampled within 6 days after treatment. Interestingly, a significant peak in plasma free 17,20ß-P levels occurred in both males and females at day 8 after treatment. Histological sections of the ovaries in these females contained oocytes at the migrating germinal vesicle stage, suggesting the role of this hormone as a maturation-inducing steroid in Atlantic bluefin tuna. In conclusion, the GnRHa implants activated effectively the reproductive endocrine axis in captive Atlantic bluefin tuna broodstocks, through stimulation of sustained elevations in plasma LH, which in turn evoked the synthesis and secretion of the relevant sex steroids leading to gamete maturation and release.

Prospective study of a new dynamic stabilisation system in the treatment of degenerative discopathy and instability of the lumbar spine.

INTRODUCTION: Posterior dynamic stabilisation (PDS) aims at relieving lumbar discogenic pain and preserving adjacent levels from accelerated degeneration. PURPOSE: To evaluate the results of a novel PDS system in 32 adult patients affected by chronic low back pain (CLBP) due to degenerative lumbar spine instability (DLSI). METHOD: A progressive follow-up for 12 months of 32 patients, with collection of complete clinical (ODI and VAS back + leg) and radiological data (resting + functional radiographs and MRI). RESULTS: Mean ODI scores improved from 49 to 6%, VAS back from 5 to 1 and VAS leg from 7 to 2. Twenty-two patients underwent fusion of a lower lumbar segment and stabilisation of an upper segment (hybrid fusion) whereas ten underwent dynamic stabilisation. In 16/32 patients, decompression was added to treat radicular pain. Motion in non-fused instrumented levels was unrestricted on functional X-rays and MRIs did not show significant morphologic changes. Four patients (12.5%) had unchanged functional and pain scores while two (6.3%) suffered worsening low back pain necessitating implant removal and spinal fusion. No infection, no new neurologic deficit or implant failure was recorded. CONCLUSIONS: The 1 year follow-up shows that the tested PDS system is able to provide a significant improvement in pain and disability scores when applied to patients affected by DLSI. The system does not provide better clinical results when compared to similar trials on posterior fusion. Further follow-up is ongoing to investigate the potential preservation of adjacent levels from accelerated degeneration.

The impact of COVID-19 and the economic crisis on Lebanese public health: Food insecurity and healthcare disintegration.

Background: The economic crisis in Lebanon, a middle-income eastern Mediterranean country, has been threatening the health of the local population. This review will look at the impact of the economic crisis and COVID-19 on health and healthcare in the country, discussing food insecurity and water shortages, and the hospital crisis for what concerns medications, electricity shortages and workforce issues. Methodology: Peer Reviewed Literature produced between 2015 and 2021, indexed in Pubmed, Scopus and Google Scholar was used to compile this short report. News and governmental reports, alongside reports of NGOs like Médicins sans frontières were also collected; these were analysed for the production of this short report. Results: The challenges and public health consequences caused by the economic crisis and the COVID-19 pandemic in Lebanon were identified and commented upon. From food insecurity and water shortages, to the Beirut port explosion and the 2021 lack of fuel and electricity, it was found that the health and well-being of the Lebanese population is currently being threatened from many points of view. With food inflation rates rapidly escalating in 2020 and peaking at 441% in October, new-born and infant milk being non-existent and 20 hour power cuts daily, the situation in Lebanon does not seem to be improving. The country needs to receive international help to relief the population from these synergetic crises. Conclusion: Long-term economic reforms with an emphasis on employment should be at the forefront of the government's priority list. International help should also be provided to prevent disasters like food insecurity and electricity shortages from posing threats to the lives and the wellbeing of the people in Lebanon again.

Long-term results of the direct repair of spondylolisthesis.

Direct repair of spondylolisthesis can save a functional segment in young patients with slight slipping. Since 1968 many surgeons have proposed different technical solutions to obtain the isthmic repair. Their results changed according to the technique used, the extent of listhesis and the age of the patient. The aim of our study was to perform a retrospective review on the long-term results of the direct repair of spondylolisthesis, according to the different techniques used. We operated 62 patients for isthmic repair, with three different techniques, from 1994 to 2007. We analysed the clinical and radiographic results of 52 cases, with an average follow-up of 9 ± 3 years (range 2-15). Ten patients were lost to the follow-up. The results were different depending on the technique used. Good or excellent clinical outcome by Odom's criteria were observed in the 83.3% of patients operated with the modified Scott technique. These results are better than those obtained in the group of patients operated with the Scott (62.5%) and the Buck technique (28.5%). Patients with clinical and radiological failure, who then underwent spinal fusion, were 57% with the Buck technique, 12.5% with the Scott technique and 2.7% with the Scott modified technique. The reasons for a new operation were symptomatic pseudarthrosis and progression of slipping. In conclusion, the pars defect repair is a helpful technique in lumbar spondylolisthesis, especially in young patient with slight slipping and painful symptoms resistant to conservative treatment. In our experience, the modified Scott technique seems to provide a better outcome than the Scott and Buck techniques.

Evidence that severe acute stress and starvation induce rapid atresia of ovarian vitellogenic follicles in Atlantic bluefin tuna, Thunnus thynnus (L.) (Osteichthyes: Scombridae).

The effects of different stressors on the atretic degeneration of ovarian vitellogenic follicles, as well as on the ovarian mass, were examined in female Atlantic bluefin tuna, Thunnus thynnus (L.), from the Mediterranean Sea. The stressors taken into consideration were short-term starvation (up to 14 days), long-term cage rearing (1 year) and crowding-induced severe panic frenzy. Wild-caught individuals were used as a control group. Fish subjected to either severe panic frenzy or starvation exhibited a decrease in gonad mass and had significantly higher intensity of α atresia in the vitellogenic follicles (means: 78% and 58%, respectively; range: 36-100%) than either wild or long-term caged individuals (means: 32% and 30%, respectively; range: 19-44%). The extensive atresia in fish stressed by severe panic frenzy was observed as early as 24 h after the stressing event. The present study represents the first evidence of the extreme susceptibility of Atlantic bluefin tuna to severe acute stress during vitellogenesis; it also shows that starvation is associated with progressive reabsorption of vitellogenic oocytes.

A histological investigation of the occurrence of non-reproductive female bluefin tuna Thunnus thynnus in the Mediterranean Sea.

The presence of non-reproductive Atlantic bluefin tuna Thunnus thynnus females in the Mediterranean Sea was investigated through histological analysis of the gonads. Three hundred and twenty-six ovary samples were collected from adults captured at different locations in the Mediterranean Sea during the reproductive seasons between 1998 and 2008. Only three specimens were considered to be in a non-reproductive state: two of them were in a reabsorbing state showing ovaries with early vitellogenic oocytes and extensive alpha and beta atresia of vitellogenic follicles; the third showed gonads with perinucleolar oocytes and was considered to be in a resting state. The low occurrence of non-reproductive individuals found in this study makes it unlikely that non-reproductive individuals aggregate with reproductive ones during their migration towards spawning grounds. Further research is suggested in order to investigate the potential presence of non-reproductive individuals on non-spawning grounds during the reproductive season.

Brain morphology and immunohistochemical localization of the gonadotropin-releasing hormone in the bluefin tuna, Thunnus thynnus.

The present study was focused on the morphology of the diencephalic nuclei (likely involved in reproductive functions) as well as on the distribution of GnRH (gonadotropin-releasing hormone) in the rhinencephalon, telencephalon and the diencephalon of the brain of bluefin tuna (Thunnus thynnus) by means of immunohistochemistry. Bluefin tuna has an encephalization quotient (QE) similar to that of other large pelagic fish. Its brain exhibits well-developed optic tecta and corpus cerebelli. The diencephalic neuron cell bodies involved in reproductive functions are grouped in two main nuclei: the nucleus preopticus-periventricularis and the nucleus lateralis tuberis. The nucleus preopticus-periventricularis consists of the nucleus periventricularis and the nucleus preopticus consisting of a few sparse multipolar neurons in the rostral part and numerous cells closely packed and arranged in several layers in its aboral part. The nucleus lateralis tuberis is located in the ventral-lateral area of the diencephalon and is made up of a number of large multipolar neurones. Four different polyclonal primary antibodies against salmon (s)GnRH, chicken (c)GnRH-II (cGnRH-II 675, cGnRH-II 6) and sea bream (sb)GnRH were employed in the immunohistochemical experiments. No immunoreactive structures were found with anti sbGnRH serum. sGnRH and cGnRH-II antisera revealed immunoreactivity in the perikarya of the olfactory bulbs, preopticus-periventricular nucleus, oculomotor nucleus and midbrain tegmentum. The nucleus lateralis tuberis showed immunostaining only with anti-sGnRH serum. Nerve fibres immunoreactive to cGnRH and sGnRH sera were found in the olfactory bulbs, olfactory nerve and neurohypophysis. The significance of the distribution of the GnRH-immunoreactive neuronal structures is discussed.

Immunohistochemical localization of CYP1A, vitellogenin and Zona radiata proteins in the liver of swordfish (Xiphias gladius L.) taken from the Mediterranean Sea, South Atlantic, South Western Indian and Central North Pacific Oceans.

Cytochrome P4501A (CYP1A) monoxygenase, vitellogenin (Vtg) and Zona radiata proteins (Zrp) are frequently used as biomarkers of fish exposure to organic contaminants. In this work, swordfish liver sections obtained from the Mediterranean Sea, the South African coasts (South Atlantic and South Western Indian Oceans) and the Central North Pacific Ocean were immunostained with antisera against CYP1A, Zrp, and Vtg. CYP1A induction was found in hepatocytes, epithelium of the biliary ductus and the endothelium of large blood vessels of fish from the Mediterranean Sea and South African waters, but not from the Pacific Ocean. Zrp and Vtg were immunolocalized in hepatocytes of male swordfish from the Mediterranean Sea and from South African waters. Plasma Dot-Blot analysis, performed in Mediterranean and Pacific specimens, revealed the presence of Zrp and Vtg in males from Mediterranean but not from Pacific. These results confirm previous findings about the potential exposure of Mediterranean swordfish to endocrine, disrupting chemicals and raise questions concerning the possible presence of xenobiotic contaminants off the Southern coasts of South Africa in both the South Atlantic and South Western Indian Oceans.

Differential lectin binding patterns in the oviductal ampulla of the horse during oestrus.

We investigated the oligosaccharide sequence of glycoconjugates, mainly sialoglycoconjugates, in the horse oviductal ampulla during oestrus by means of lectin and pre-lectin methods such as the KOH-neuraminidase procedure to remove sialic acid residues and incubation with N-glycosidase F to cleave N-linked glycans. Ciliated cells displayed N-linked oligosaccharides throughout the cytoplasm. The cilia glycocalyx expressed both N- and O-linked (mucin-type) oligosaccharides, both showing a high variety of terminal sequences. In the most non-ciliated cells, the whole cytoplasm contained N-linked oligosaccharides with terminal alphaGal as well as mucin-type glycans with terminal Forssman pentasaccharides. In a few scattered non-ciliated cells, the whole cytoplasm displayed sialylated N-linked oligosaccharides with terminal Neu5Ac-GalNAc and O-linked glycans terminating with neutral and/or alphaGalNAc, Neu5Ac alpha2,6Gal/GalNAc, Neu5AcGal beta1,3GalNAc. Supra-nuclear granules, probably Golgi zones, of non-ciliated cells showed mainly O-linked glycans rich in sialic acid residues. The luminal surface of non-ciliated cells showed N-linked oligosaccharides, containing terminal/internal alphaMan/alphaGlc, betaGlcNAc and terminal alphaGal, as well as mucin-type oligosaccharides terminating with a large variety of either neutral saccharides or sialylated sequences. Apical protrusions containing O-linked oligosaccharides with terminal Forssman pentasaccharide, Neu5Ac-Gal beta1,4GlcNAc, Neu5Ac-GalNAc were seen in non-ciliated cells scattered along the epithelium. These findings show the presence of sialoglycoconjugates in the oviductal ampulla epithelium of the mare and the existence of different lectin binding profiles between ciliated and non-ciliated (secretory) cells, as well as the presence of non-ciliated cell sub-types which might determine functional differences along the ampullary epithelium of mare oviduct.

Paraspinal approach to the far lateral disc herniations: retrospective study on 42 cases.

Forty-two patients underwent surgery for far-lateral disc herniations. Average patient age was 45.1 years, 28 patients were male and 14 female. The level concerned most was L4-5 disc (55%). A paramedian muscle-splitting intertransverse approach is described for this type of disc herniation. This method allows direct visualization of the disc and root involved and does not provide bone resection and important surgical manipulation. Ninety-one percent of the patients reported excellent or good results according to Macnab outcome criteria with an average follow-up of 32.5 months (range 1-5 years). Twelve patients (28.6%) developed postoperative transient radicular pain that disappeared in 15-21 days after analgesic therapy. There were no recurrences or spinal instability. The paramedian muscle-splitting intertransverse approach is a rational technique. Its advantages are that the spine is not opened and spinal stability is maintained. It requires minimal soft-tissue and bone resection and the herniated disc is directly visualized. Moreover, it contains minimal manipulation of the neuro-vascular structures and avoids significant muscle retraction. However, it requires an adequate learning curve and good familiarity with microsurgical techniques.

Glycoconjugates during the annual sexual cycle in lizard epididymal ductuli efferentes: a histochemical study.

The epididymal ductuli efferentes of the lizard Podarcis sicula campestris De Betta are lined with simple, columnar, nonciliated and ciliated cells. The use of lectin histochemistry has provided information about changes of sugars associated with glycoconjugates of epithelial cells and intraluminal spermatozoa which are conveyed from the longitudinal canal to the cranial region of the ductus epididymis. Epithelial cells exhibited residues of alpha-D-mannose, N-acetylglucosamine, and beta-D-galactose-(1-4)-N-acetylglucosamine as revealed with lectins Con A, WGA and RCA120, respectively, throughout the sexual cycle. An increase of RCA120 staining was observed on microvilli of nonciliated cells and in the cytoplasm of ciliated cells during the reproductive period. However, during the following refractory period, when the organ was in regression, there was a decreased staining with Con A on microvilli and the absorbent surface of nonciliated cells, with WGA in nonciliated cells and the cytoplasm of ciliated cells, and with RCA120 on microvilli and the cytoplasm of both cell types. Terminal alpha-D-galactose residues were increasingly stained from autumn up to the reproductive period with BS I-B4 on microvilli, the absorbent surface and cilia, whereas they were entirely lacking during the refractory period. UEA I revealed alpha-L-fucose residues on the absorbent surface of nonciliated cells during the abortive and reproductive periods, increasing in the latter period when cilia also expressed this sugar. Terminal alpha/beta-D-N-acetylgalactosamine was evidenced with SBA on the absorbent surface of nonciliated cells during the reproductive period. The terminal beta-D-galactose-(1-3)-N-acetylgalactosamine dimer was never found with PNA, whereas O- and N-linked sialoglycoconjugates were present only during the reproductive period. The spermatozoa head exhibited N-linked glycans with high-mannose content, and beta-D-galactose-(1-4)-N-acetylglucosamine as well as O- and N-linked sialoglycoconjugates throughout the year. During the reproductive period, oligosaccharides with alpha-D-mannose residues increased, and oligosaccharides with terminal alpha-D-galactose, alpha-L-fucose and sialic acid-N-acetylgalactosamine dimer were also present. Unlike spermatozoa of seminiferous tubules, the spermatozoa head of the lizard epididymal ductuli efferentes exhibited seasonal variability in the lectin binding pattern which may be related to time-dependent changes in the glycoconjugate profiles of epithelial cells.