Brain washing and neural health: role of age, sleep, and the cerebrospinal fluid melatonin rhythm.

The brain lacks a classic lymphatic drainage system. How it is cleansed of damaged proteins, cellular debris, and molecular by-products has remained a mystery for decades. Recent discoveries have identified a hybrid system that includes cerebrospinal fluid (CSF)-filled perivascular spaces and classic lymph vessels in the dural covering of the brain and spinal cord that functionally cooperate to remove toxic and non-functional trash from the brain. These two components functioning together are referred to as the glymphatic system. We propose that the high levels of melatonin secreted by the pineal gland directly into the CSF play a role in flushing pathological molecules such as amyloid-ß peptide (Aß) from the brain via this network. Melatonin is a sleep-promoting agent, with waste clearance from the CNS being highest especially during slow wave sleep. Melatonin is also a potent and versatile antioxidant that prevents neural accumulation of oxidatively-damaged molecules which contribute to neurological decline. Due to its feedback actions on the suprachiasmatic nucleus, CSF melatonin rhythm functions to maintain optimal circadian rhythmicity, which is also critical for preserving neurocognitive health. Melatonin levels drop dramatically in the frail aged, potentially contributing to neurological failure and dementia. Melatonin supplementation in animal models of Alzheimer's disease (AD) defers Aß accumulation, enhances its clearance from the CNS, and prolongs animal survival. In AD patients, preliminary data show that melatonin use reduces neurobehavioral signs such as sundowning. Finally, melatonin controls the mitotic activity of neural stem cells in the subventricular zone, suggesting its involvement in neuronal renewal.

Neurotensin Release from Dopamine Neurons Drives Long-Term Depression of Substantia Nigra Dopamine Signaling.

Midbrain dopamine neurons play central physiological roles in voluntary movement, reward learning, and motivated behavior. Inhibitory signaling at somatodendritic dopamine D2 receptor (D2R) synapses modulates excitability of dopamine neurons. The neuropeptide neurotensin is expressed by many inputs to the midbrain and induces LTD of D2R synaptic currents (LTDDA); however, the source of neurotensin that is responsible for LTDDA is not known. Here we show, in brain slices from male and female mice, that LTDDA is driven by neurotensin released by dopamine neurons themselves. Optogenetic stimulation of dopamine neurons was sufficient to induce LTDDA in the substantia nigra, but not the VTA, and was dependent on neurotensin receptor signaling, postsynaptic calcium, and vacuolar-type H+-ATPase activity in the postsynaptic cell. These findings reveal a novel form of signaling between dopamine neurons involving release of the peptide neurotensin, which may act as a feedforward mechanism to increase dopamine neuron excitability.SIGNIFICANCE STATEMENT Dopamine neurons in the midbrain play a critical role in reward learning and the initiation of movement. Aberrant dopamine neuron function is implicated in a range of diseases and disorders, including Parkinson's disease, schizophrenia, obesity, and substance use disorders. D2 receptor-mediated PSCs are produced by a rare form of dendrodendritic synaptic transmission between dopamine neurons. These D2 receptor-mediated PSCs undergo LTD following application of the neuropeptide neurotensin. Here we show that release of neurotensin by dopamine neurons themselves is sufficient to induce LTD of dopamine transmission in the substantia nigra. Neurotensin signaling therefore mediates a second form of interdopamine neuron communication and may provide a mechanism by which dopamine neurons maintain excitability when nigral dopamine is elevated.

Melatonin: highlighting its use as a potential treatment for SARS-CoV-2 infection.

Numerous pharmaceutical drugs have been repurposed for use as treatments for COVID-19 disease. These drugs have not consistently demonstrated high efficacy in preventing or treating this serious condition and all have side effects to differing degrees. We encourage the continued consideration of the use of the antioxidant and anti-inflammatory agent, melatonin, as a countermeasure to a SARS-CoV-2 infection. More than 140 scientific publications have identified melatonin as a likely useful agent to treat this disease. Moreover, the publications cited provide the rationale for the use of melatonin as a prophylactic agent against this condition. Melatonin has pan-antiviral effects and it diminishes the severity of viral infections and reduces the death of animals infected with numerous different viruses, including three different coronaviruses. Network analyses, which compared drugs used to treat SARS-CoV-2 in humans, also predicted that melatonin would be the most effective agent for preventing/treating COVID-19. Finally, when seriously infected COVID-19 patients were treated with melatonin, either alone or in combination with other medications, these treatments reduced the severity of infection, lowered the death rate, and shortened the duration of hospitalization. Melatonin's ability to arrest SARS-CoV-2 infections may reduce health care exhaustion by limiting the need for hospitalization. Importantly, melatonin has a high safety profile over a wide range of doses and lacks significant toxicity. Some molecular processes by which melatonin resists a SARS-CoV-2 infection are summarized. The authors believe that all available, potentially beneficial drugs, including melatonin, that lack toxicity should be used in pandemics such as that caused by SARS-CoV-2.

Melatonin in ventricular and subarachnoid cerebrospinal fluid: Its function in the neural glymphatic network and biological significance for neurocognitive health.

The central nervous system (CNS) is endowed with a specialized cerebrospinal fluid (CSF)/lymph network which removes toxic molecules and metabolic by-products from the neural parenchyma; collectively, this has been named the glymphatic system. It allows CSF located in the subarachnoid space which surrounds the CNS to enter the depths of the brain and spinal cord by means of Virchow-Robin perivascular and perivenous spaces. CSF in the periarterial spaces is transferred across the astrocytic end feet which line these spaces aided by AQ4 channels; in the interstitium, the fluid moves via convection through the parenchyma to be eventually discharged into the perivenous spaces. As it passes through the neural tissue, the interstitial fluid flushes metabolic by-products and extracellular toxins and debris into the CSF of the perivenous spaces. The fluid then moves to the surface of the CNS where the contaminants are absorbed into true lymphatic vessels in the dura mater from where it is shunted out of the cranial vault to the cervical lymph nodes. Pineal melatonin released directly into the CSF causes the concentration of this molecule to be much higher in the CSF of the third ventricle than in the blood. After the ventricular melatonin enters the subarachnoid and Virchow-Robin spaces it is taken into the neural tissue where it functions as a potent antioxidant and anti-inflammatory agent. Experimental evidence indicates that it removes pathogenic toxins, e.g., amyloid-ß and others, from the brain to protect against neurocognitive decline. Melatonin levels drop markedly during aging, coincident with the development of several neurodegenerative diseases and the accumulation of the associated neurotoxins.

Sirtuins and the circadian clock interplay in cardioprotection: focus on sirtuin 1.

Chronic disruption of circadian rhythms which include intricate molecular transcription-translation feedback loops of evolutionarily conserved clock genes has serious health consequences and negatively affects cardiovascular physiology. Sirtuins (SIRTs) are nuclear, cytoplasmic and mitochondrial histone deacetylases that influence the circadian clock with clock-controlled oscillatory protein, NAMPT, and its metabolite NAD+. Sirtuins are linked to the multi-organ protective role of melatonin, particularly in acute kidney injury and in cardiovascular diseases, where melatonin, via upregulation of SIRT1 expression, inhibits the apoptotic pathway. This review focuses on SIRT1, an NAD+-dependent class III histone deacetylase which counterbalances the intrinsic histone acetyltransferase activity of one of the clock genes, CLOCK. SIRT1 is involved in the development of cardiomyocytes, regulation of voltage-gated cardiac sodium ion channels via deacetylation, prevention of atherosclerotic plaque formation in the cardiovascular system, protection against oxidative damage and anti-thrombotic actions. Overall, SIRT1 has a see-saw effect on cardioprotection, with low levels being cardioprotective and higher levels leading to cardiac hypertrophy.

Triple variation in the vascular supply in the neck of a female donor body: a case report.

PURPOSE: Understanding the anatomical variations involving bifurcation of the common carotid artery, positioning of external and internal carotid arteries, and branching of the external carotid artery are of vital importance in neck surgeries such as carotid endarterectomies (CEA). METHODS: The neck of a 51-year-old female donor body was dissected to demonstrate the arterial network. RESULTS: Bifurcation of the common carotid artery occurred at the level of the C6-C7 intervertebral disc, significantly inferior to the generally accepted and taught anatomical location at the level of intervertebral disc between C3 and C4 vertebrae. When the arteries were followed superiorly after the bifurcation, a unique second variation was observed: translocation of the external and internal carotid arteries. The external carotid artery was located posterolaterally and the internal carotid artery was located more medially. Finally, a third variation was discovered in the form of a common thyrolingual trunk that gave rise to superior thyroid and lingual arteries rather than these arising independently from the external carotid artery. CONCLUSIONS: We report a unique triple variation within the major arteries of the neck that has not been previously reported in surveyed literature. This case report may provide useful information for cardiovascular surgeons performing CEA and for otolaryngologists performing prophylactic arterial ligation following transoral robotic surgery for oropharyngeal cancer resection.

Melatonin in Mitochondria: Mitigating Clear and Present Dangers.

In cancer cells, glucose is primarily metabolized to pyruvate and then to lactate in the cytosol. By allowing the conversion of pyruvate to acetyl-CoA in mitochondria, melatonin reprograms glucose metabolism in cancer cells to a normal cell phenotype. Acetyl-CoA in the mitochondria also serves as a necessary co-factor for the rate-limiting enzyme in melatonin synthesis, thus ensuring melatonin production in mitochondria of normal cells.

Switching diseased cells from cytosolic aerobic glycolysis to mitochondrial oxidative phosphorylation: A metabolic rhythm regulated by melatonin?

This commentary reviews the concept of the circadian melatonin rhythm playing an essential role in reducing the development of diseases such as solid tumors which adopt cytosolic aerobic glycolysis (Warburg effect) to support their enhanced metabolism. Experimental data show that solid mammary tumors depend on aerobic glycolysis during the day but likely revert to mitochondrial oxidative phosphorylation at night for ATP production. This conversion of diseased cells during the day to a healthier phenotype at night occurs under control of the circulating melatonin rhythm. When the nocturnal melatonin rise is inhibited by light exposure at night, cancer cells function in the diseased state 24/7. The ability of melatonin to switch cancer cells as well as other diseased cells, for example, Alzheimer disease, fibrosis, hyperactivation of macrophages, etc, from aerobic glycolysis to mitochondrial oxidative phosphorylation may be a basic protective mechanism to reduce pathologies.

Central and peripheral actions of melatonin on reproduction in seasonal and continuous breeding mammals.

Under field conditions, especially for mammals that inhabit high latitudes, the regulation of seasonal breeding activity to ensure delivery of the young at the time most conducive to their survival is essential. This is most frequently accomplished by the annual reproductive cycle being linked to seasonal photoperiod changes which determine the nocturnal duration of the pineal melatonin signal. Mating can occur during any season that ensures spring/early summer delivery of the offspring. Thus, the season of mating is determined by the duration of pregnancy. The precise hormonal control of the annual cycle of reproduction by melatonin is accomplished at the level of the hypothalamo-pituitary axis which, in turn, determines the physiological state of the gonad and adnexa due to the regulation of pituitary gonadotrophin release. Many species are continuous rather than seasonal breeders. In these species, melatonin has a minor hormonal influence on the central regulation of reproduction but, nevertheless, its antioxidant functions at the level of the gonads support optimal reproductive physiology. Possibly like all cells, those in the ovary, e.g., granulosa cells and oocytes (less is known about melatonin synthesis by the testes or spermatogenic cells), synthesize melatonin which is used locally to combat free radicals and reactive nitrogen species which would otherwise cause oxidative/nitrosative stress to these critically important cells. Oxidative damage to the oocyte, zygote, blastocyst, etc., results in an abnormal fetus which is either sloughed or gives rise to an unhealthy offspring. The importance of the protection of the gametes (both oocytes and sperm) from oxidative molecular mutilation cannot be overstated. Fortunately, as a highly effective free radical scavenger and indirect antioxidant (by upregulating antioxidant enzyme), locally-produced melatonin is in the optimal location to protect the reproductive system from such damage.

Melatonin inhibits Warburg-dependent cancer by redirecting glucose oxidation to the mitochondria: a mechanistic hypothesis.

Melatonin has the ability to intervene in the initiation, progression and metastasis of some experimental cancers. A large variety of potential mechanisms have been advanced to describe the metabolic and molecular events associated with melatonin's interactions with cancer cells. There is one metabolic perturbation that is common to a large number of solid tumors and accounts for the ability of cancer cells to actively proliferate, avoid apoptosis, and readily metastasize, i.e., they use cytosolic aerobic glycolysis (the Warburg effect) to rapidly generate the necessary ATP required for the high metabolic demands of the cancer cells. There are several drugs, referred to as glycolytic agents, that cause cancer cells to abandon aerobic glycolysis and shift to the more conventional mitochondrial oxidative phosphorylation for ATP synthesis as in normal cells. In doing so, glycolytic agents also inhibit cancer growth. Herein, we hypothesize that melatonin also functions as an inhibitor of cytosolic glycolysis in cancer cells using mechanisms, i.e., downregulation of the enzyme (pyruvate dehydrogenase kinase) that interferes with the conversion of pyruvate to acetyl CoA in the mitochondria, as do other glycolytic drugs. In doing so, melatonin halts the proliferative activity of cancer cells, reduces their metastatic potential and causes them to more readily undergo apoptosis. This hypothesis is discussed in relation to the previously published reports. Whereas melatonin is synthesized in the mitochondria of normal cells, we hypothesize that this synthetic capability is not present in cancer cell mitochondria because of the depressed acetyl CoA; acetyl CoA is necessary for the rate limiting enzyme in melatonin synthesis, arylalkylamine-N-acetyltransferase. Finally, the ability of melatonin to switch glucose oxidation from the cytosol to the mitochondria also explains how tumors that become resistant to conventional chemotherapies are re-sensitized to the same treatment when melatonin is applied.

Neurotensin receptor 1 deletion decreases methamphetamine self-administration and the associated reduction in dopamine cell firing.

We previously reported that a non-selective pharmacological blockade of neurotensin receptors in the ventral tegmental area (VTA) decreases methamphetamine (METH) self-administration in mice. Here, we explored the consequences of genetic deletion of neurotensin receptor 1 (NtsR1) on METH self-administration and VTA dopamine neuron firing activity. We implanted mice with an indwelling jugular catheter and trained them to nose-poke for intravenous infusions of METH. Mice with NtsR1 deletion (KO) acquired self-administration similar to wildtype (WT) and heterozygous (HET) littermates. However, in NtsR1 KO and HET mice, METH intake and motivated METH seeking decreased when the response requirement was increased to a fixed ratio 3 and when mice were tested on a progressive ratio protocol. After completion of METH self-administration, single cell in vivo extracellular recordings of dopamine firing activity in the VTA were obtained in anesthetized mice. Non-bursting dopamine neurons from KO mice fired at slower rates than those from WT mice, supporting an excitatory role for NtsR1 on VTA dopamine neuronal activity. In WT mice, a history of METH self-administration decreased dopamine cell firing frequency compared with cells from drug-naïve controls. NtsR1 KO and HET mice did not exhibit this decline in dopamine cell firing activity after METH experience. We also observed an increase in population activity following METH self-administration that was strongest in the WT group. Our results suggest a role for NtsR1 in METH-seeking behavior and indicate that ablation of NtsR1 prevents the detrimental effects of prolonged METH self-administration on VTA dopamine cell firing frequency.

Anti-Warburg Effect of Melatonin: A Proposed Mechanism to Explain its Inhibition of Multiple Diseases.

Glucose is an essential nutrient for every cell but its metabolic fate depends on cellular phenotype. Normally, the product of cytosolic glycolysis, pyruvate, is transported into mitochondria and irreversibly converted to acetyl coenzyme A by pyruvate dehydrogenase complex (PDC). In some pathological cells, however, pyruvate transport into the mitochondria is blocked due to the inhibition of PDC by pyruvate dehydrogenase kinase. This altered metabolism is referred to as aerobic glycolysis (Warburg effect) and is common in solid tumors and in other pathological cells. Switching from mitochondrial oxidative phosphorylation to aerobic glycolysis provides diseased cells with advantages because of the rapid production of ATP and the activation of pentose phosphate pathway (PPP) which provides nucleotides required for elevated cellular metabolism. Molecules, called glycolytics, inhibit aerobic glycolysis and convert cells to a healthier phenotype. Glycolytics often function by inhibiting hypoxia-inducible factor-1α leading to PDC disinhibition allowing for intramitochondrial conversion of pyruvate into acetyl coenzyme A. Melatonin is a glycolytic which converts diseased cells to the healthier phenotype. Herein we propose that melatonin's function as a glycolytic explains its actions in inhibiting a variety of diseases. Thus, the common denominator is melatonin's action in switching the metabolic phenotype of cells.

Melatonin and Pathological Cell Interactions: Mitochondrial Glucose Processing in Cancer Cells.

Melatonin is synthesized in the pineal gland at night. Since melatonin is produced in the mitochondria of all other cells in a non-circadian manner, the amount synthesized by the pineal gland is less than 5% of the total. Melatonin produced in mitochondria influences glucose metabolism in all cells. Many pathological cells adopt aerobic glycolysis (Warburg effect) in which pyruvate is excluded from the mitochondria and remains in the cytosol where it is metabolized to lactate. The entrance of pyruvate into the mitochondria of healthy cells allows it to be irreversibly decarboxylated by pyruvate dehydrogenase (PDH) to acetyl coenzyme A (acetyl-CoA). The exclusion of pyruvate from the mitochondria in pathological cells prevents the generation of acetyl-CoA from pyruvate. This is relevant to mitochondrial melatonin production, as acetyl-CoA is a required co-substrate/co-factor for melatonin synthesis. When PDH is inhibited during aerobic glycolysis or during intracellular hypoxia, the deficiency of acetyl-CoA likely prevents mitochondrial melatonin synthesis. When cells experiencing aerobic glycolysis or hypoxia with a diminished level of acetyl-CoA are supplemented with melatonin or receive it from another endogenous source (pineal-derived), pathological cells convert to a more normal phenotype and support the transport of pyruvate into the mitochondria, thereby re-establishing a healthier mitochondrial metabolic physiology.

Collateral circulation of the femoral and genicular systems in human lower limbs is highly uncommon.

The descending branch of the lateral circumflex artery is a septocutaneous vessel that is vital for free and pedicle thigh flap transfer surgeries when repairing tissue defects. It also forms an anastomosis with the superior lateral genicular artery to create a collateral pathway for circumventing occlusions in the superficial femoral artery (SFA). Many anatomical texts and atlases imply the persistence of this anastomosis. However, previous studies indicate variability in the source of the arteries that form the anastomosis, and have reported cases where an anastomosis does not exist. We hypothesized that variations from the conventional accepted pattern can be predicted by comparisons of arterial diameters, and that unconventional anastomoses may be present to facilitate collateral circulation to the limb. Fifty-one limbs were dissected and analyzed to establish the source of the descending branch of the lateral circumflex artery, classify the types of anastomoses, and compare the diameters of the descending branch of the lateral circumflex artery, the SFA and the profunda femoris artery to the common femoral artery (CFA). Vessel diameters were normalized to the diameter of the CFA to allow comparison of limbs from both sexes and to minimize the effects of cadaver size on correlating vessel size to the presence or absence of collateral circuits. We report that 62.7% of limbs (32/51) had typical branching patterns; however, only 27.4% of limbs (14/51) had any anastomosis to connect the proximal and distal regions of the thigh. Importantly, the SFA had a wider relative diameter in limbs without anastomoses than in limbs that had normal anastomoses, perhaps precluding the formation of a collateral pathway. Overall, collateral circulation of the lower limb was highly uncommon, in contrast to information inferred from anatomical texts. This study suggests the need for more thorough procedures for determining viable anastomoses prior to thigh flap surgeries to ensure flap survival.

Case report of unique anastomosis between facial and inferior alveolar arteries.

PURPOSE: Understanding anatomical variations of the facial artery and its branches is important for dental and medical practitioners. METHODS: Routine cadaveric dissection of the head and neck was performed to demonstrate the origin and branches of the facial artery. RESULTS: Facial artery emerged from a common linguofacial trunk off the external carotid artery. On the face, the facial artery first gave off a pre-masseteric branch. Immediately after, an aberrant artery emerged from the facial artery that coursed along the ramus of the mandible, which upon further dissection and examination was found to anastomose with inferior alveolar artery within the ramus of the mandible. CONCLUSIONS: We report a unique anastomosis between facial and inferior alveolar arteries, vessels that have not been previously shown to communicate. This case report may provide useful information for oral and maxillofacial surgeons as well as dentists performing inferior alveolar nerve blocks.

Dopaminergic Neurons Exhibit an Age-Dependent Decline in Electrophysiological Parameters in the MitoPark Mouse Model of Parkinson's Disease.

Dopaminergic neurons of the substantia nigra (SN) play a vital role in everyday tasks, such as reward-related behavior and voluntary movement, and excessive loss of these neurons is a primary hallmark of Parkinson's disease (PD). Mitochondrial dysfunction has long been implicated in PD and many animal models induce parkinsonian features by disrupting mitochondrial function. MitoPark mice are a recently developed genetic model of PD that lacks the gene for mitochondrial transcription factor A specifically in dopaminergic neurons. This model mimics many distinct characteristics of PD including progressive and selective loss of SN dopamine neurons, motor deficits that are improved byl-DOPA, and development of inclusion bodies. Here, we used brain slice electrophysiology to construct a timeline of functional decline in SN dopaminergic neurons from MitoPark mice. Dopaminergic neurons from MitoPark mice exhibited decreased cell capacitance and increased input resistance that became more severe with age. Pacemaker firing regularity was disrupted in MitoPark mice and ion channel conductances associated with firing were decreased. Additionally, dopaminergic neurons from MitoPark mice showed a progressive decrease of endogenous dopamine levels, decreased dopamine release, and smaller D2 dopamine receptor-mediated outward currents. Interestingly, expression of ion channel subunits associated with impulse activity (Cav1.2, Cav1.3, HCN1, Nav1.2, and NavB3) was upregulated in older MitoPark mice. The results describe alterations in intrinsic and synaptic properties of dopaminergic neurons in MitoPark mice occurring at ages both before and concurrent with motor impairment. These findings may help inform future investigations into treatment targets for prodromal PD. SIGNIFICANCE STATEMENT: Parkinson's disease (PD) is the second most diagnosed neurodegenerative disorder, and the classic motor symptoms of the disease are attributed to selective loss of dopaminergic neurons of the substantia nigra. The MitoPark mouse is a genetic model of PD that mimics many of the key characteristics of the disease and enables the study of progressive neurodegeneration in parkinsonism. Here we have identified functional deficits in the ion channel physiology of dopaminergic neurons from MitoPark mice that both precede and are concurrent with the time course of behavioral symptomatology. Because PD is a progressive disease with a long asymptomatic phase, identification of early functional adaptations could lay the groundwork to test therapeutic interventions that halt or reverse disease progression.

Aging decreases L-type calcium channel currents and pacemaker firing fidelity in substantia nigra dopamine neurons.

Substantia nigra dopamine neurons are involved in behavioral processes that include cognition, reward learning, and voluntary movement. Selective deterioration of these neurons is responsible for the motor deficits associated with Parkinson's disease (PD). Aging is the leading risk factor for PD, suggesting that adaptations occurring in dopamine neurons during normal aging may predispose individuals to the development of PD. Previous studies suggest that the unique set of ion conductances that drive spontaneous, rhythmic firing of action potentials could predispose substantia nigra dopamine neurons to selective neurodegeneration. Here we show, using patch-clamp electrophysiological recordings in brain slices, that substantia nigra dopamine neurons from mice 25-30 months of age (old) have comparable membrane capacitance and input resistance to neurons from mice 2-7 months of age (young). However, neurons from old mice exhibit slower firing rates, narrower spike widths, and more variable interspike intervals compared with neurons from young mice. Dopamine neurons from old mice also exhibit smaller L-type calcium channel currents, providing a plausible mechanism that likely contributes to the changes in impulse activity. Age-related decrements in the physiological function of dopamine neurons could contribute to the decrease in voluntary movement and other dopamine-mediated behaviors observed in aging populations. Furthermore, as pharmacological antagonism of L-type calcium channels has been proposed as a potential treatment for the early stages of PD, our results could point to a limited temporal window of opportunity for this therapeutic intervention.

Differentiated adipose-derived stem cell cocultures for bone regeneration in polymer scaffolds in vivo.

Critical-sized bone defects can lead to significant morbidity, and interventions are limited by the availability and donor-site morbidity of bone grafts. Polymer scaffolds seeded with cells have been explored to replace bone grafts. Adipose-derived stem cells have shown great promise for vascularization and osteogenesis of these constructs, and cocultures of differentiated stem cells are being explored to augment vessel and bone formation. Adipose-derived stem cells were differentiated into endothelial cells and osteoblasts, and in vitro studies showed increased proliferation of cocultured cells compared with undifferentiated adipose-derived stem cells and monocultures of endothelial cells and osteoblasts. The cells were seeded into polylactic acid gas-plasma-treated scaffolds as cocultures and monocultures and then implanted into critical-sized rat calvarial defects. The cocultures were in a 1:1 osteoblast to endothelial cell ratio. The increase in proliferation seen by the cocultured cells in vitro did not translate to increased vascularization and osteogenesis in vivo. In vivo, there were trends of increased vascularization in the endothelial cell group and increased osteogenesis in the osteoblast and endothelial monoculture groups, but no increase was seen in the coculture group compared with the undifferentiated adipose-derived stem cells. Endothelial cells enhance vascularization and osteoblast and endothelial cell monocultures enhance bone formation in the polymer scaffold. Predifferentiation of adipose-derived stem cells is promising for improving vascularization and osteogenesis in polymer scaffolds but requires future evaluation of coculture ratios to fully characterize this response.

Food is Medicine Initiative for Mitigating Food Insecurity in the United States.

OBJECTIVES: While several food assistance programs in the United States tackle food insecurity, a relatively new program, "Food is Medicine," (FIM) initiated in some cities not only addresses food insecurity but also targets chronic diseases by customizing the food delivered to its recipients. This review describes federal programs providing food assistance and evaluates the various sub-programs categorized under the FIM initiative. METHODS: A literature search was conducted from July 7, 2023 to November 9, 2023 using the search term, "Food is Medicine", to identify articles indexed within three major electronic databases, PubMed, Medline, and Cumulative Index to Nursing and Allied Health Literature (CINAHL). Eligibility criteria for inclusion were: focus on any aspect of the FIM initiative within the United States, and publication as a peer-reviewed journal article in the English language. A total of 180 articles were retrieved; publications outside the eligibility criteria and duplicates were excluded for a final list of 72 publications. Supporting publications related to food insecurity, governmental and organizational websites related to FIM and other programs discussed in this review were also included. RESULTS: The FIM program includes medically tailored meals, medically tailored groceries, and produce prescriptions. Data suggest that it has lowered food insecurity, promoted better management of health, improved health outcomes, and has, therefore, lowered healthcare costs. CONCLUSIONS: Overall, this umbrella program is having a positive impact on communities that have been offered and participate in this program. Limitations and challenges that need to be overcome to ensure its success are discussed.