Twelve-day Period Research Articles

First Detection and Characterization of Smacovirus in the Human Vagina in Two Sequential Samples over a Twelve-Day Interval

Background: Smacovirus is a CRESS-DNA virus identified almost exclusively in transient fecal samples from various vertebrate species. Objective: We evaluated human vaginal samples for the presence and maintenance of Smacovirus. Methods: Viral metagenomics analysis was performed on vaginal samples collected from 28 apparently healthy women in New York City, USA. Twenty-one of the women provided duplicate samples over a 12–21-day interval. Results: Phylogenetic analysis identified two samples from the same individual, collected over a twelve-day interval, that were positive for the complete Smacovirus genome. All detected sequence contigs belonged to a single variant of CRESS-DNA. Conclusions: The continuous presence of Smacovirus in the human vagina over a twelve-day period was identified.

Time-dependent changes in striatal monoamine levels and gene expression following single and repeated amphetamine administration in rats

As drug addiction may result from pathological usurpations of learning and memory's neural mechanisms, we focused on the amphetamine-induced time-dependent neurochemical changes associated with neural plasticity. We used juvenile rats as the risk for drug abuse is higher during adolescence. Experiment 1 served to define the appropriate amphetamine dose and the neurochemical effects of a single administration. In experiment 2, rats received seven amphetamine or saline injections in the open-field test throughout a twelve-day period. We measured the mRNA levels of the brain-derived neurotrophic factor (BDNF), its tropomyosin receptor kinase B (TrkB), the cAMP response element-binding protein (CREB), the microRNA-132, the Rho GTPase-activating protein 32 (p250GAP), the corticotropin-releasing factor (CRF), and monoamines and amino-acids contents in the nucleus accumbens and the dorsal striatum 45, 90, and 180 min after the last injection. We found that amphetamine changed gene expression only at certain time points and in a dose and region-dependent manner. Repeated but not single administrations upregulated accumbal and striatal BDNF (180 min) and striatal pri-miR-132 (90 min) expression, while downregulated accumbal CREB levels (90 min). As only some drug users develop addiction, we compared brain parameters between low and high amphetamine responders. Prone subjects characterized by having reduced striatal 5-HT metabolism, higher accumbal BDNF and TrkB expression, and lower levels of CREB in the dorsal striatum and p250GAP in both regions. Thus, individual differences in drug-induced changes in neurotransmission and gene expression in nigrostriatal and mesolimbic dopaminergic pathways may underlie the plasticity adaptations associated with behavioral sensitization to amphetamine.

Suspected cyanide toxicity in cattle associated with ingestion of laurel - a case report

BackgroundCyanide is one of the most rapidly acting toxins affecting cattle, with poisoning typically occurring following ingestion of cyanogenic plants. Laurel (Prunus laurocerasus), is one such potentially toxic cyanogenic plant. This case report details fatalities in an Irish herd following the ingestion of laurel and aims to raise awareness of the potential risk that access to laurel hedges poses to farm animals.Case presentationOver a twelve-day period, the death occurred of 36 dairy-cross weanlings; the majority (22 weanlings) died over a two-day period. Two days following entry to a field bounded by a laurel hedge, the weanlings displayed signs of lethargy and profuse green diarrhoea. In the majority of animals there was a limited response to treatment with antimicrobials, vitamin B complex and fluid therapy. Recumbency and death ensued. Cyanosis was noted terminally. Two weanlings were submitted for post mortem examination. Laurel leaves were identified in the rumen contents of one weanling. Post mortem findings and additional test results on cohort animals suggested a number of pathological processes may have been involved in the animals, possibly complicating/exacerbating the effects of laurel ingestion. However, cyanide was considered a factor in a least some of the casualties and arrangements were made to test for cyanide on blood samples from a random selection of seven cohort animals. Although collected one week after exposure to the laurel hedge, toxic cyanide levels were recorded in a sample from one animal, which subsequently died.ConclusionsThe large fatality rate serves as a timely reminder to include plant poisoning as a differential diagnosis when dealing with large numbers of rapid fatalities. Failure to thoroughly examine rumen contents and collect a detailed history in this case, could easily have allowed death to be attributed to other causes and the involvement of cyanide toxicity to be missed. In cases of individual or group fatalities, history is invaluable and recent entry to new grazing areas or any potential diet change or access to garden plants should be thoroughly investigated.

Parental Preference for Treatment Location and Inclusion in Decision Making in Painful Pediatric Procedures: Towards an Evidence-Based Clinical Guideline

PurposeTo determine parental preference for treatment location of painful procedures for pediatric patients, and the involvement of parents in decision-making about procedure location. Design and methodA descriptive survey design was applied. Parents of hospitalized children were asked what procedures their child had undergone, where the procedures were performed, who decided on the location, what their preference was, and why. Data was collected over a twelve-day period in seven wards of a tertiary pediatric hospital. ResultsOf 312 different procedures, 244 (78%) were performed in the patient's room. The treatment room was reserved for complex procedures such as lumbar punctures. Seventy-three percent of parents surveyed indicated a strong preference for procedures to be performed in the patient's room, with the most common reason being ‘comfortability’. Parents were only involved in procedure location decision-making in 50% of the reported cases. ConclusionsFor parents, the patient's room is the preferred location for many procedures. The opinion of parents was not always taken into consideration by health care professionals, in contrary to the principles of patient-centered care. These findings are important for health care professionals in pediatric settings to reconsider the location of procedures and how parents are involved in decision making.

The combined therapy of a traditional Chinese medicine formula and Western medicine for a critically ill case infected with COVID-19

ObjectivePresentation of a case illustrating the benefits of traditional Chinese medicine (TCM) for treatment of Coronavirus disease 2019 (COVID-19) in critically ill patients. Clinical features and outcomeA 58-year-old woman presented with cough, fever, dizziness, chest tightness, polypnea and poor appetite. She was admitted to Guizhou Provincial People’s hospital, and diagnosed with critically ill type of COVID-19 in February 2020. According to the patient's symptoms and signs, the TCM syndrome differentiation was qi deficiency, dampness-stasis and toxin accumulation. Then she received the combined therapy of a modified Chinese herbal formula and Western medicine. During a twelve-day period of treatment, her respiratory distress and appetite quickly improved. Abnormal laboratory indicators were resumed in time and lung lesions in CT scan largely absorbed. No side effects associated with this Chinese herbal formula were found. Before discharge, two consecutive nasopharyngeal swabs were shown to be negative for severe acute respiratory coronavirus 2 (SARS-CoV-2). ConclusionsOur case report suggests that collaborative treatments with traditional Chinese medicine prove beneficial in the management of COVID-19 in critically ill patients. In order to give optimal care for this COVID-19 crisis for the whole world, Chinese medicine practitioners and Western medical doctors should work together in frontline.

Dugesia japonica is the best suited of three planarian species for high-throughput toxicology screening

High-throughput screening (HTS) using new approach methods is revolutionizing toxicology. Asexual freshwater planarians are a promising invertebrate model for neurotoxicity HTS because their diverse behaviors can be used as quantitative readouts of neuronal function. Currently, three planarian species are commonly used in toxicology research: Dugesia japonica, Schmidtea mediterranea, and Girardia tigrina. However, only D. japonica has been demonstrated to be suitable for HTS. Here, we assess the two other species for HTS suitability by direct comparison with D. japonica. Through quantitative assessments of morphology and multiple behaviors, we assayed the effects of 4 common solvents (DMSO, ethanol, methanol, ethyl acetate) and a negative control (sorbitol) on neurodevelopment. Each chemical was screened blind at 5 concentrations at two time points over a twelve-day period. We obtained two main results: First, G. tigrina and S. mediterranea planarians showed significantly reduced movement compared to D. japonica under HTS conditions, due to decreased health over time and lack of movement under red lighting, respectively. This made it difficult to obtain meaningful readouts from these species. Second, we observed species differences in sensitivity to the solvents, suggesting that care must be taken when extrapolating chemical effects across planarian species. Overall, our data show that D. japonica is best suited for behavioral HTS given the limitations of the other species. Standardizing which planarian species is used in neurotoxicity screening will facilitate data comparisons across research groups and accelerate the application of this promising invertebrate system for first-tier chemical HTS, helping streamline toxicology testing.

The Effect of Musical Intervention Vs Standard Treatment on Reducing Agitation Over a Twelve-Day Period

The Effect of Musical Intervention Vs Standard Treatment on Reducing Agitation Over a Twelve-Day Period

Preformulation of a liquid dosage formulation of captopril for pediatric use: drug-excipient compatibility and stability studies

Currently, medications used in children are typically modified from pharmaceutical dosage forms designed for adults. Captopril is widely adapted to liquid formulations for use in hospitals. Its stability in the aqueous medium is reduced since it undergoes oxidation producing captopril disulfide (its main metabolite). The aim of this formulation study was to suggest favorable conditions for the development of a stable captopril formulation. The compatibility between the drug and excipients was evaluated by differential scanning calorimetry analysis (DSC). For studies in solution, different formulations were prepared according to a factorial design varying EDTA concentration, water purity and pH. The resultant formulations were stored at 60°C and analyzed over a twelve-day period using HPLC. The DSC curves obtained suggested, although not conclusive to elucidation, interactions of captopril with citric acid and sucralose. The stability study of these solutions revealed that the variables significantly influenced captopril content, which degraded at zero order kinetics and rates differing by a factor of up to 7 times, where pH proved the most influential factor. Interactions between variables were observed. Therefore, development of a stable captopril formulation is feasible provided EDTA and a buffering agent is used at suitable concentrations (0.08% and pH 3.85).

Effect of two commercial probiotic products on population growth of rotifer Brachionus rotundiformis Tschugunoff

The application of probiotics has varying effects on rotifer culture. These effects can either be directly on their growth and reproduction or indirectly by affecting the culture environment. The role played by specific probiotic candidates is still indistinct and further research is required to ascertain their significance in rotifer culture. In this study, euryhaline rotifer Brachionus rotundiformis (SS-type) was used to evaluate the effect of two commercial probiotic products on rotifer density, reproductive parameters and unionized ammonia concentration for a twelve-day culture period. To estimate these parameters, rotifers were cultured in glass bottles containing dilute sterilized sea water with the following five diet treatments: Nannochloropsis oculata (control), Igsign (PB1), Toaraze Aqua (PB2) or the combination of N. oculata with either PB1 or PB2. The rotifer density was significantly affected by the addition of probiotic. N. oculata + PB2 treatment resulted in higher rotifer density and specific growth rate compared to the control treatment. Mixis induction was unaffected by N. oculata + PB2 while it was repressed by N. oculata + PB1. Ammonia concentration was not significantly affected with the addition of probiotics. These results indicate that probiotics have synergetic effects with microalgae diet that resulted in the highest rotifer densities.

Measurement and modeling of the spatiotemporal dynamics of beach surface moisture content

Abstract The spatiotemporal dynamics of surface moisture exert a significant influence on the operation of aeolian transport systems at many beaches. However, we currently lack the detailed understanding of variability in surface moisture required to incorporate it into aeolian models. This problem is addressed here through direct measurements and simulation modeling of beach surface moisture over a twelve-day period, and through quantification of the relative contributions of evaporation, condensation and groundwater inputs to surface moisture. It was found that the beach surface can be characterized spatially in terms of three moisture zones: a consistently dry zone ( 40%). The relative influence of groundwater inputs was found to decrease moving landward, as the depth of the water table increased and the amplitude of tidally-induced water table fluctuations decreased. The critical pressure head (groundwater depth) at which evaporation begins to impose a demonstrable influence on surface moisture variability was found to be 90–100 cm. Temporally, beach surface moisture is a function of the lunar tidal cycle at longer-term time scales (weekly), and diurnal tidal and evaporation cycles. A numerical model was used to simulate capillary transfers of moisture from the water table to the surface, and moisture losses due to evaporation. The model was found to reliably replicate the measured spatiotemporal variability in surface moisture. In the dry zone, where most aeolian transport would be expected to occur, simulated moisture contents were typically within ±0.2% of measured volumetric contents.

The President and the Autopen: It is Unconstitutional for Someone or Something to Sign a Bill Outside of the President's Presence

On May 26, 2011, only hours before three provisions of the Foreign Intelligence Surveillance Act were scheduled to expire, Congress passed an extension. For days, the White House had someone ready to fly to Europe with the legislation in hand for the President to sign, but Congress had been tardy. It seemed quite important to the White House that none of these provisions lapse for any length of time, even the relatively short time it would take to fly from Washington to France. With this urgency as a backdrop, the President was awakened at 5:45 a.m. Central European Time so he could authorize a first: phone a White House staffer in Washington, D.C., and instruct him to use an auto-pen to sign the bill. No President had ever had anyone else sign on his behalf, and certainly no President had ever ordered the use of an auto-pen to inscribe his signature on a bill when away from the White House. This Article explores multiple facets of the President’s use of a proxy signature to sign legislation. The state of the law surrounding proxy signatures has remained amazingly constant through both English and American history. The proxy and the principal must be present together when a proxy signature is utilized for a high-value transaction. This was the rock-solid law when the constitution was written.No one seems focused on the presence requirement: not the President, nor the Republican House members who complained to the President, nor the numerous legal commentators and scholars that we have heard from since the auto-pen signing occurred. The use of the auto-pen, itself unproblematic, seems to have eclipsed the more important issue of whether the president was present when it was used.By definition, if a non-presence proxy signature is affixed to a bill, then time is of the essence. When time is of the essence, the consequences of a voided signature are never greater. Since the President has roughly twelve days to sign a bill, a non-presence proxy signature could produce one of two unintended consequences. At best, it would produce a twelve-day period during which the new legislation would not be in effect. At worst, if Congress is not in session at the end of the twelve-day period, it would amount to a pocket veto. Neither case is consistent with well-ordered government.In sum, the President’s use of the auto-pen (or even a human being) to sign a bill outside of the President’s presence is unconstitutional. This establishes a dangerous precedent, one which every thinking lawyer in Washington politics seemed to have overlooked. Let the auto-pen episode not be a precedent for this or any other President to follow. The Constitution does not allow for shortcuts.

Methane and ammonia emissions from a beef feedlot in western Canada for a twelve-day period in the fall

Commercial feedlot operations are becoming a mainstay in the Canadian beef industry. These large operations that typically raise thousands of animals at a time represent a large localized source of methane (CH4) and of atmospheric pollutants such as ammonia (NH3) and particulate matter. An inverse dispersion model was utilized to calculate CH4 and NH3 emissions from a commercial cattle feedlot and an adjacent runoff retention pond. The feedlot measurements were collected within the interior of the feedlot enabling a near continuous emissions record over the 12 d of the study period. Average daily emission estimates of CH4 and NH3 were 323 and 318 g animal-1 d-1, respectively. The CH4 emissions represent 4% of the gross energy intake (GEI) and NH3 emissions represent 72% of the total N intake. Emissions from the runoff retention pond associated directly with the feedlot operation were approximately 2.7 and 2% of the daily average feedlot emissions of CH4 and NH3, respectively. Key words: Methane, ammonia, inverse dispersion, ruminant livestock, trace gas emissions

Cortical Neuromodulation by Constraint-Induced Movement Therapy in Congenital Hemiparesis: An fMRI Study

The aim of this study was to assess neuromodulative effects of CIMT in congenital hemiparesis. Ten patients (age range: 10-30 years) with congenital hemiparesis due to unilateral cortico-subcortical infarctions in the middle cerebral artery territory, and with preserved cortico-spinal projections from the affected hemisphere to the paretic hand, were included. After a twelve-day period of constraint-induced movement therapy (CIMT), all showed a significant improvement of paretic hand function. Immediately before and after therapy, functional MRI during active and passive hand movements was performed to monitor cortical activation. Four patients showed consistent increases in cortical activation during movements of the paretic hand in the primary sensorimotor cortex of the affected hemisphere. Of the remaining six patients, three showed similar changes, but these results were potentially contaminated by an improved task performance after therapy. No significant alteration in activation was observed in two patients, and one showed movement artifacts. Even a short period of CIMT can induce changes of cortical activation in congenital hemiparesis. In our sample, increases in fMRI activation were consistently observed in the primary sensorimotor cortex of the affected hemisphere. Thus, the potential for neuromodulation is preserved in the affected hemisphere after early brain lesions.

Light microscope studies on coccolithophorids from the western Mediterranean Sea, with notes on combination cells of Daktylethra pirus and Syracosphaera pulchra

ABSTRACT Distribution and abundance of phytoplankton in the western Mediterranean were studied during a twelve-day period in the early spring of 1961. A total of 38 species of coccolithophorids belonging to 30 genera and 8 families, as well as an association between the heterococcolithophorid Syracosphaera pulchra and the holococcolitho-phorid Daktylethra pirus were identified and counted. Only Emiliania huxleyi was numerically abundant. A higher diversity was found at stations influenced by Atlantic water along the Tunisian-Sicilian coast and in the Tyrrhenian Sea than at sites located towards the North. Holococcolithophorids formed 34% of the total number of species recorded. Members of the two families Syracosphaeraceae (24%) and Rhabdosphaeraceae (21%) dominated, compared with Coccolithaceae (8%) and Noëlaerhabdaceae (5%). A list of confirmed species as well as original photomicrographs and transmission electron micrographs are presented.

Bottom Boundary Layer Parameters and Sediment Transport on the Louisiana Inner-Shelf During Cold Front Passages

ABSTRACT Increasing focus has been placed on the critical role the inner-shelf plays in coastal sediment dynamics. During storms, bottom sediment may be mobilized by waves and currents and transported along or across the inner-shelf, affecting the sediment budget of the adjacent coast. During a twelve-day period that included two cold front passages, we measured waves and near-bed currents on the Louisiana inner-shelf (depth ~ 8m) using a sophisticated bottom-mounted instrumentation system. We then calculated bottom boundary layer parameters using wave-current interaction models, including those of Grant and Madsen (1986) and Glenn and Grant (1987). Finally, we predicted sediment transport by assuming steady state turbulent diffusion within and above the wave boundary layer. Results indicate that the second front (Event 2) was the more energetic of the two, and maximum significant wave height (1.33 m), and current speed (0.22 ms-1), occurred during this time. Additionally, mean current-induced shear velocity (1.9 cms-1) and wave-current shear velocity (4.4 cms-1) were highest during this event's frontal and prefrontal stages (respectively). During the postfrontal stage, currents were strong and well organized although combined shear velocities were low as a result of reduced wave height. Predicted sediment transport varied considerably in direction and magnitude throughout the deployment, but was highest (0.13-0.14 gcm-1s-1 SE) during the prefrontal and frontal stages of Event 2. Fair weather transport was low and to the west. We conclude that cold fronts are an important mechanism for sediment movement on the Louisiana inner-shelf, although the associated transport direction and magnitude require further quantification.

Antioxidant defence of red blood cells and plasma in stored human blood

Blood collected routinely from donors, and preserved with CPDA-1 anticoagulant (citrate, phosphate, dextrose, adenine), was investigated. The concentration of reduced glutathione, and glutathione peroxidase, glutathione S-transferase, glutathione reductase, superoxide dismutase, and catalase activities in erythrocytes, as well as the total radical-trapping antioxidant parameter in plasma were determined on days 1, 3, 7, 12, 16, 20 and 25 of storage. At the end of the study, a 30% decrease in the reduced glutathione concentration ( P<0.001) and decreases in glutathione S-transferase (over 20%, P<0.001), glutathione reductase (over 8%, P<0.01) and superoxide dismutase (over 10%, P<0.001) activities in erythrocytes, together with up to a 30% diminution of total antioxidant activity in plasma ( P<0.001) were noted. Thus, during blood storage, glutathione-dependent antioxidant systems in erythrocytes and antioxidant defence in plasma are depleted. From the present study, a twelve-day period can be considered a safe storage limit. The sequence of events occurring in stored blood, leading to peroxidative injury in erythrocytes, is discussed.

Healing of laser-induced sclerostomy channels in the rabbit

Obliteration of sclerostomy canals during the course of healing is of course undesirable, but before interference with the process can hope to be successful, a stepwise ultrastructural delineation of the course of events provoked by surgucal intervention is essential. In the current study, such an analysis is undertaken, and the changes in tissue morphology discerned correlated with observed modulations in IOP. Two cw-Nd:YAG laser sclerostomies were created ab interno on one eye in each of ten rabbits; the unoperated fellow-eyes served as controls. IOP was monitored daily over a twelve-day period. Changes incurred within the canals and to collaterally damaged scleral tissue were analyzed at two-day intervals by light and electron microscopy. The development of well-defined filtering blebs demonstrated the success of the procedure. IOP was significantly lowered during the entire course of the observation period, but after the fifth postoperative day, the blebs had disappeared. Within five days of surgical intervention, morphological analysis revealed the canal to be invaded by macrophages originating from both the iris root and episcleral tissue; these were actively engaged in the phagocytosis of intensely damaged collagen abutting on the lumen. A few days later, the lumen had become occluded by fibroblasts and a dense capillary network. The course of regeneration observed within scleral tissue which had undergone moderate thermal insult, suggests that collagen fibrils undergo a process of repolymerization. Although the time course of repair is more rapid in rabbits than in humans, the data gleaned nonetheless yield valid information respecting the sequence of events involved in the scarification process. The loose nature of the tissue occluding the canal lumen apparently permits the percolation of fluid through it, thus accounting for the clinical discrepancy between a continuing decrease in IOP and the disappearance of a filtering bleb.

In vitro cultivation of cartilage tissue for reconstructive surgery: effect of L(+)-lactate and glycolate on cultivated human chondrocytes

Within the scope of producing cartilage tissue in a three-dimensional culture design, the stability of the used delivery substance in-vitro tissue product has to be improved. For this, carrier materials consisting of bioresorbable polymers, e. g. poly(L[+]-lactic acid) and poly(glycolic acid) can be used. In respect of the biocompatibility of these polymers, the effect of degradation products on chondrocytes is of major interest. The available biomaterials were tested on chondrocytes in form of their monomers, glycolic acid and L(+)-lactic acid. Effects in regard of cell activity were determined with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide( MTT)test. A non-pH-effect was examined by buffering with concentrated NaOH. In a short-term testing with increasing monomer concentrations as well as in a test over a twelve-day period, L(+)-lactic acid proved to have a lower cytotoxic effect on chondrocytes than glycolic acid. Similar results were obtained with buffered culture media. Therefore, poly(L[+]-lactic acid) can be recommended for the development of chondrocytes-polymer constructs for in-vitro engineering of cartilage tissue.

Differential Inhibition of Photosynthesis ( in vivo and in vitro) and Changes in Chlorophyll a Fluorescence Induction Kinetics of four Tobacco Cultivars under Drought Stress

Summary This study was conducted in an attempt to explain the observed difference in drought tolerance of four cultivars of Nicotiana tabacum L., by monitoring plant water status and several aspects of photosynthesis, including chlorophyll a fluorescence, under controlled environmental conditions. Drought stress of increasing intensity, ranging from light to severe, was induced over a twelve-day period by withholding water. Photosynthetic oxygen evolution was monitored in vivo at a CO 2 concentration exceeding 5 % to exclude any stomatal limitation and was expressed as quantum efficiency (Φ). Partial photochemical reactions were determined polarographically with isolated thylakoids of the drought-stressed tissue. From the Kautsky curves the I/P-ratio and ΔF-values were determined at various decreasing leaf water potentials (Ψ L ). All four cultivars exhibited a drought stress-induced decline in Φ, PSI-, PSII- and PSI + PSII-activity, but the extent and nature differed among the different cultivars. The Φ-values of the drought-tolerant cultivars were characterized by a fast initial decline that stabilized as the Ψ L declined, whereas the Φ-values of drought-sensitive cultivars did not show a similar response, but declined at a slow continuous rate as the drought stress intensified. The PSII-activity and PSI- + PSII-activity were influenced in much the same way. PSII-activity was found to be more sensitive than PSI-activity and even more so in the case of the drought-sensitive than in the drought-tolerant cultivars. It was concluded that the observed stress-induced decline in Φ and PSI- + PSII-activity could largely be ascribed to an inhibition of PSII-activity. This conclusion is corroborated by both the fluorescence data regarding the I/P-ratio and ΔF-values, of which the former was found to increase earlier and reach higher end-values in the drought-sensitive cultivars, and the latter, which indicated that the drought-tolerant cultivars were characterized by a higher photosynthetic efficiency at low Ψ L . A mechanism is proposed which might retard or reverse drought stress induced stromal acidification and might explain the apparently higher photosynthetic dehydration tolerance of the drought-tolerant cultivars. The possible involvement of ABA is discussed.

The Dark Days and the Light Month

The actual calendars of Indo-European peoples do show intercalary additions to the lunar year but these take the form of intercalary months. Frazer was aware of this, saying that 'there are grounds for thinking that at a very early time the Aryan peoples sought to correct their lunar year, not by inserting twelve supplementary days every year, but by allowing .the annual deficiency to accumulate for several years and then supplying it by a whole intercalary month.' 2 He expressed his puzzlement over this in connection with the Celts of Gaul, commenting: 'Why they abandoned the simple and obvious expedient of annually intercalating twelve days, and adopted instead the more recondite system of intercalating a month of thirty days every two and a half years, is not plain.'3 The answer I have to offer is that the Indo-Europeans never did employ a twelve-day intercalary period but kept the months of the lunar year in line with the natural seasons by inserting intercalary months when required, and that the place of the twelve days in the calendar was not outside the lunar year but inside it. As I see it, the possibility of having a group of twelve days that stand in a functional relationship to the rest of the year arises when the month used in the calendar is not the synodic month of twenty-nine or thirty days but the shorter light month, the period of the moon's visibility. Ten years before Frazer published his remarks, W. H. Roscher had studied the matter of month length in connection with Greek weeks and festivals and had concluded that months of two different lengths-27-8 days and 29-30 days-were known in antiquity, but that the use of the short month of 27-8 days was the older.4 Frazer made no mention of this finding in his discussion of the twelve days, and yet the two ideas of the light month and a set of special days can be seen as complementary and I shall consider them together here. The longest period of the moon's visibility is twenty-eight days and the moon goes through its cycle from new moon, through full moon, to the last crescent of the waning moon in this time. As Plutarch expressed it, referring to the number twenty-eight: 'Such is the number of the moon's illuminations and in so many days does it revolve through its own cycle.'5 Between each pair of light months there is a gap which was called by the Romans the 'interlunium' or 'intermenstruum,' that is, the interval between the moons or the months.6 The treatment of this time of the dark of the moon