This variability was due to large trial-to-trial variations in the response of most individual neurons (Bartho et al., 2009; Hromádka et al., 2008). Despite the variability, we were able to observe

that sound intensity and identity modulated the probability of observing a population event. Tuning to pure tones could be seen at both the single neuron (Figures 2B–2D) and at the population level (Figure 2A). However, prediction of the population firing rate in response to complex sounds by a linear model based on the observed pure tone tuning was poor (Figure S2). Therefore, local populations encode sounds in a nonlinear fashion, as was reported for single neurons (Machens et al., 2004). This implies that pure tone tuning alone is not sufficient to describe sound representation in the auditory cortex. We therefore decided to use a more general framework to investigate MK-1775 mw the coding properties of local response patterns in single trials (Bathellier et al., 2008). In most local populations, we made the striking observation that despite the high

variability of response patterns, the most reliable part of the pattern seemed to be common to very different sounds GS-1101 nmr such as the different pure tones shown in Figure 2A. This suggested to us that sound evoked responses in local auditory cortex networks are constrained to a limited repertoire of functional patterns superposed on high trial-to-trial stochasticity. To obtain a quantitative account of the limited repertoire of functional patterns in the face of large variability, we systematically quantified the similarity of local response patterns elicited by large arrays of short (50–70 ms) pure tones and complex

sounds. To do so, we used a similarity metric designed to obtain an intuitive readout of single trial response separability. In short, the similarity between two sound-evoked responses was defined as the average of all pairwise correlations between the single trial response patterns of the two sounds (see matrices of single trial correlations, Figure 3A). This similarity metric was compared to a response reliability metric, which was the average of all pairwise correlations between all the single trial response Rebamipide patterns of one given sound. This reliability metric gave us a quantitative readout of the trial-to-trial variability in response to a given sound. Using these two metrics, the idea is that if the response patterns to two sounds have a lower similarity than their respective reliabilities, they will likely be discriminable on a single trial basis by an external observer. If not, the patterns can be thought to be the same. Pairwise response similarities were displayed in color-coded matrix plots after ordering the sounds with a hierarchical clustering algorithm to reveal potential underlying structures in the space of response patterns ( Figures 3B and 3C).

Mutation of site 1 produced a −4.5 kcal/mol loss of binding energy for the GluR6Δ2F58A homodimer, but only a −2.9 kcal/mol loss for the GluR6Δ2F58A/KA2Y57A heterodimer, with equal contributions by the F58A and Y57A mutants, of −1.4 and −1.5 kcal/mol, respectively

(Table S1). The excess total dimerization energy, totaling 3.1 kcal/mol, must come from other sites in the heterodimer interface. The mutation E156A produced a loss of only −0.43 kcal/mol; for L163A the loss was −1.31 kcal/mol. The S165G/T168A double mutant produced a loss of binding energy of −1.74 kcal/mol. Strikingly, Ser165 and Thr168 do not make contacts with the GluR6 subunit, and instead merely serve to stabilize the loop conformation which positions Leu163 and Ile164 in the dimer interface. To examine whether the binding Pexidartinib order mechanism for heterodimer formation was an additive Buparlisib or cooperative process we performed a mutant cycle analysis looking at interactions between sites in domains R1 and R2 with both the GluR6Δ2 and GluR6Δ2F58A mutant used as heterodimer partners. Mutant cycles were calculated as shown in Figure 5B, where coupling coefficients (Ω) greater than one indicate positive cooperativity (Carter

et al., 1984 and Hidalgo and MacKinnon, 1995). The analysis yielded coupling coefficients (Ω values) of only 0.8–1.7 and reveals clearly that heterodimer assembly is an additive process with little cooperativity between domains R1 and R2 (Supplemental Experimental Procedures).

Sodium butyrate The much larger disruption of heterodimer assembly observed for the I164A mutant likely reflects conformational changes resulting from destabilization of the hydrophobic patch formed by the loop rearrangement in the KA2 subunit. Of note, amino acid sequence alignments (Figure 5C) reveal that in other iGluR subunits Ile164 is replaced by charged or polar residues, consistent with a unique role for Ile164 in mediating heterodimer assembly for KA1 and KA2. This alignment also reveals exchange of Glu167 (Asp165 in KA1) by Trp/Ile/Leu in other iGluR subunits (Figure 5C). The residues exchanged are in a flexible loop region connecting helix F and strand 7, the conformation of which differs in individual iGluR families. In GluR1–4 and GluR5–7 the Trp/Ile/Leu residues form part of the hydrophobic core of domain 2, while in the KA2 subunit the polar residues are surface exposed, and make intersubunit contacts in the heterodimer assembly. In the KA1 and KA2 subunits, Phe160/162 fills the space in the hydrophobic core which in other iGluR subunits is occupied by the Trp/Ile/Leu residues which align with Asp165/Glu167 in KA1 and KA2. At the corresponding position in the AMPA receptors and GluR5–7 the Phe residue is replaced by smaller Ala or Pro side chains. In order to elucidate the structure of the GluR6/KA2 ATD tetramer, we crystallized a complex of wt GluR6 and KA2.

In summary, 0.2 mL of egg suspension, containing approximately IWR 1 100 eggs, was added per well, in 24-well plates. They were incubated for 24 h at 27 °C to obtain L1. After egg hatching, 90 μL of culture medium (containing Escherichia coli and yeast extract) was added to each well, followed by the test extract. The concentrations were determined following the ratio of 2 from 5 μg mL−1 to 0.0003 μg mL−1 (i.e., 5, 2.5, 1.25 μg mL−1 and so on) in at least six replicates for each concentration. The highest

and the lowest concentrations evaluated for each plant extract were as follows: 0.313–0.0195 mg mL−1 for P. tuberculatum, 0.0195–0.0003 mg mL−1 for L. sidoides, 0.156–0.0098 mg mL−1 for M. piperita, 2.5–0.0078 mg mL−1

for H. crepitans and 5–0.039 mg mL−1 for C. guianensis. The plates were incubated for 6 days at 25 °C, and then the differential count from L3, L2 and L1 was performed. A solution of 0.5% DMSO was prepared as negative control and 0.64 mg L−1 of ivermectin as positive control, in six replicates. In both in vitro tests, the lowest concentration was determined when the XAV-939 price hatching and larval development were similar to the control. The highest concentration was based on the solubility or on the turbidity that limits its readability. Sixty Wistar rats (Rattus norvergicus) were infected by subcutaneous inoculation with 2000 infective larvae of S. venezuelensis. The production of infective larvae, animal inoculation and determination of parasite load followed the descriptions of Nakai and Amarante

(2001). Seven days after infection, the animals were divided into six groups (n = 10) to receive treatment by gavage as follows: G1 – positive control (Albendazole – 10 mg kg−1); G2 – negative control (Sorbitol – 100%); G3 – P. tuberculatum extract (150 mg kg−1); G4 – P. tuberculatum extract (250 mg kg−1); G5 – L. sidoides essential oil (150 mg kg−1) and G6 – L. sidoides Parvulin essential oil (250 mg kg−1). Infection intensity was determined by counting the number of eggs per gram of feces (EPG) on days 1, 2, 3, 4 and 6 after the first day of treatment and by counting the number of parthenogenetic female worms found in the first-third portion of the small intestine. To obtain adult worms, the upper third of the small intestine was removed from the rats 13 days after infection, cut longitudinally and incubated in saline solution (0.9% NaCl) for 4 h at 39 °C ( Nakai and Amarante, 2001). The aim of this test was to evaluate in rats the extracts that showed greatest activity in the in vitro tests. Thus, it would be possible to obtain an indication of the most active extract in an in vivo model for future testing in sheep. Although the essential oil of M. piperita showed good results in the in vitro tests, it was not possible to perform the in vivo test with it due to the small amount of extract that was provided by the partner institution.

The timeframe for this correlated well with neurite sprouting and synapse stabilization based on previous studies of developing connections in vitro (Soriano et al., 2008). However, in α-syn-hWT pff-treated WT, but not α-syn −/− neurons, the maturation of functional connections never reached the level achieved in PBS-treated cultures, as a significant reduction was observed 10 days after α-syn-hWT pff Fulvestrant treatment (Figure 8F). This functional connectivity was severely compromised 14 days after treatment and the network consisted of just a few sparse connections at this time point

(Figures 8E and 8F). In summary, the formation of insoluble aggregates of endogenous α-syn results in early disruption in coordinated network activity. Later, as more α-syn inclusions develop and propagate throughout the neuron, excitatory tone is decreased and functional connectivity is greatly reduced. Here Talazoparib price we demonstrate that seeds derived from α-syn amyloid fibrils generated with pure recombinant full-length and truncated human WT α-syn, when directly

added to mouse primary hippocampal neurons, are internalized and induce the recruitment of endogenous soluble α-syn into insoluble pathologic LB-like and LN-like α-syn aggregates resembling those found in human synucleinopathies. Indeed the verisimilitude of these α-syn aggregates in cultured mouse neurons to LBs and LNs found in PD brains is striking because they are found in perikarya and extensively in neurites, insoluble, filamentous by EM and immuno-EM, hyperphosphorylated, and ubiquitinated, and they exclude β-syn (Baba et al., 1998, Duda et al., 2000, Fujiwara et al., 2002 and Spillantini et al., 1998). Notably, these aggregates initially formed within axons, sequestering endogenous α-syn away from presynaptic terminals, followed by propagation into the somata. Over time, formation of these α-syn aggregates leads to selective alterations in synaptic proteins, compromises neuronal excitability and connectivity, and culminates in neuron death. Thus, we have developed a neuronal culture model of PD-like α-syn inclusion formation that allows for the dissection of the mechanisms leading to the formation of LBs and LNs, as well as

for studies of the impact of Edoxaban these inclusions on the function and viability of affected neurons. Moreover, since the majority of PD and DLB cases are sporadic and are not caused by mutations or overexpression of α-syn, our neuronal model system provides a means to study the pathogenesis of α-syn in sporadic PD, as well as other α-synucleinopathies. We show that α-syn pffs made from pure, recombinant protein are highly potent in the recruitment of the endogenously expressed protein into LB-like and LN-like α-syn pathology, in contrast to previous studies that have relied on experimental manipulations such as protein overexpression of WT and mutant proteins, and/or extrinsic factors to introduce pffs into cells (Clavaguera et al., 2009, Frost et al., 2009, Guo and Lee, 2011 and Luk et al.

It is needless to say that calcium imaging can be successfully performed in many other species, including zebrafish (e.g., Brustein et al., 2003, Sumbre et al., 2008 and Yaksi Talazoparib nmr et al., 2009), Aplysia (e.g., Gitler and Spira, 1998), crayfish (e.g., Ravin et al., 1997), developing Xenopus (e.g., Demarque and Spitzer, 2010, Hiramoto and Cline,

2009 and Tao et al., 2001), frog (e.g., Delaney et al., 2001), squid (e.g., Smith et al., 1993), turtle (e.g., Wachowiak et al., 2002), Drosophila (e.g., Seelig et al., 2010, Wang et al., 2003 and Yu et al., 2003), blowfly (e.g., Elyada et al., 2009), and honey bee (e.g., Galizia et al., 1999). Imaging dendritic spines, the postsynaptic site of excitatory connections in many neurons, was one of the first biological applications of two-photon calcium imaging. Combining two-photon microscopy with calcium imaging in hippocampal brain slices demonstrated that calcium signals can be restricted

to dendritic spines (Figures 5Aa and 5Ab) (Yuste and Denk, 1995). The authors showed additionally that spine calcium signals were abolished by the application of the blockers of glutamatergic transmission. Subsequently, synaptically evoked spine calcium signaling was found to be caused by a variety of other mechanisms, depending on the type of neuron (Denk et al., 1995, Finch and Augustine, 1998, Kovalchuk et al., 2000, Raymond and Redman, 2006 and Wang et al., 2000). For example, Alectinib purchase Figures 5Ac and 5Ad show results obtained with confocal calcium imaging from mouse cerebellar parallel fiber-Purkinje cell synapses. The authors identified the calcium signaling mechanism of metabotropic glutamate receptor type 1-mediated transmission, involving calcium release from internal stores in dendrites and spines (Takechi et al., 1998). It has been shown that such a localized dendritic calcium signaling is essential for the induction of long-term synaptic depression (Konnerth et al., 1992 and Wang et al., 2000), a possible cellular mechanism underlying motor learning in the cerebellum (Aiba et al., 1994 and Bender et al., 2006). Similarly, the calcium dynamics at presynaptic terminals are

also accessible to calcium imaging (Delaney et al., 1989, Regehr and Tank, 1991a, Regehr and Tank, 1991b, Rusakov et al., 2004 and Smith et al., 1993). For this purpose, Liothyronine Sodium presynaptic terminals are loaded with an appropriate calcium indicator dye. A nice example is illustrated in Figures 5Ba–5Bc. To image climbing fibers in the cerebellar cortex, the authors injected the calcium indicator fluo-4 together with the morphological marker Texas red dextran upstream into the inferior olive of neonatal rats in vivo (Kreitzer et al., 2000). The dextran-conjugated calcium dye and Texas red were taken up by the inferior olive neurons and diffused within a few days through the climbing fibers to the cerebellar cortex. Thus, climbing fibers could be identified in subsequently prepared cerebellar slices.

Flies were shown different rotation stimuli (rotating square wave gratings, single dark and light edges, opposing edges) or a translational

stimulus moving either front-to-back or back-to-front. Female flies of all genotypes were tested at 34°C, a restrictive temperature for Shits activity. In vivo calcium imaging was done largely as described in Clark et al. (2011). The stimulus display was modified and stimuli were projected onto a rear-projection screen in front of the fly. Flies were shown 2 s-lasting full-field light flashes, a moving bar or a Gaussian random flicker stimulus. See Supplemental Experimental Procedures for detailed methods. We thank Nirao Shah, Liqun Luo, Christian Klämbt, David Kastner, Girish Deshpande, Proteases inhibitor Saskia de Vries, Jennifer Esch, and Tina Schwabe for critical comments on the manuscript. We thank Georg Dietzl and Sheetal Bhalerao for providing the phototaxis assay, Christoph Scheper and Ya-Hui Chou for brain dissections, and Alexander Katsov for help with the high-throughput behavioral assay. M.S. and D.A.C. acknowledge postdoctoral fellowships from the Jane Coffin Childs Memorial Fund for Medical Research. D.M.G was supported by a Ruth L. Kirschstein

NRSA Postdoctoral Fellowship (F32EY020040) from the National Eye Institute. Y.E.F. acknowledges an NIH Neuroscience Research Training grant (5 T32 MH020016-14), and L.F. was supported by a Fulbright CP-868596 purchase International Science and Technology Scholarship and a Bio-X

Stanford Interdisciplinary Graduate Fellowship (Bruce and Elizabeth Dunlevie fellow). D.A.C also received support Ketanserin from an NIH T32 Postdoctoral Training Grant. This work was funded by a National Institutes of Health Director’s Pioneer Award DP1 OD003530 (T.R.C.) and by R01 EY022638. “
“Fly motion detection is a key model system for studying fundamental principles of neural computation. Flies exhibit robust visual behaviors (Heisenberg and Wolf, 1984), and neurons in the fly visual system are highly sensitive to visual motion stimuli (Hausen, 1982). A mathematical model for visual motion detection, the Hassenstein-Reichardt elementary motion detector (HR-EMD; Hassenstein and Reichardt, 1956), successfully reconciles a wide range of behavioral and electrophysiological phenomena measured in flies (Egelhaaf and Borst, 1989, Götz, 1964, Haag et al., 2004 and Hausen and Wehrhahn, 1989). The basic operation of the HR-EMD is a multiplication of two input signals after one of them has been temporally delayed (Figure 1B; Reichardt, 1961). The “correlation-type” structure of the HR-EMD is highly similar to models for motion detection in the vertebrate retina (Borst and Euler, 2011) and may represent a common neural computation across sensory systems (Carver et al., 2008). In spite of the success of the EMD model, its cellular implementation remains unknown.

Many proteins that regulate developmental processes, e.g., neural induction and neuronal differentiation, axon growth, and synaptogenesis, are also expressed in the adult brain, serving related or different functions. A case in point is neurotrophins, a small family of nerve growth factor-related proteins (Chao,

2003 and Huang and Reichardt, 2003). While initially identified as factors that promote survival and axon growth of specific selleck chemical neuronal populations, neurotrophins have been found to regulate dendrite growth and pruning, synaptic function and plasticity, and sensory perception and cognitive processes (Park and Poo, 2013). Development of ocular dominance columns in V1 requires the action of extracellularly present brain-derived neurotrophic factor (BDNF) and activation of its TrkB

receptors (Cabelli et al., 1997) that is known to influence maturation of GABAergic inhibition (Huang et al., 1999) and potentiate excitatory synaptic functions (Poo, 2001). Aberrant neurotrophin signaling could cause both abnormal development and dysfunction of the adult brain, as suggested by human genetic association studies and the altered expression of neurotrophins and their receptors in affected brain regions in many neurological and neuropsychiatric diseases (Chao et al., 2006). A common single-nucleotide polymorphism (SNP) in the human Bdnf gene—the substitution of valine at codon 66 with methionine (V66M)—results in up to 30% reduction in the level of BDNF secretion but is genetically linked to impaired memory performances ( Egan et al., 2003 and Hariri learn more et al., 2003) and brain development ( Pezawas et al.,

2004) in humans. Mice with genetic variant BDNF (V66M) exhibited increased anxiety-related behaviors ( Chen et al., 2006) and reduced ability in motor learning ( Fritsch et al., 2010). Interestingly, transcranial Dipeptidyl peptidase direct current stimulation (tDCS) in both humans and mice resulted in enhanced motor learning and elevated BDNF level in the mice brains ( Fritsch et al., 2010). Although tDCS does not target specific circuits, anodal stimulation may provide a general enhancement of excitability (via depolarization) that helps the expression of specific activity-dependent plasticity associated with the learning process. Neural plasticity contributes to the recovery of function after brain injury. In patients with stroke, for example, there is usually some spontaneous recovery over the first several months (Cramer, 2008). Task-specific activity has also been shown to be a critical factor for promoting recovery (Nudo et al., 1996b; Ramanathan et al., 2006). After a “hand-area” stroke, intensive retraining in nonhuman primates was specifically associated with an expansion of the cortical representation for hand and digits into the previous proximal arm representation (Nudo et al.

The current model suggests that stage-specific ecdysis behavior is produced by small triggering steps (probably due to other releasing factors, including the neuropeptides corazonin (COR) (Kim et al., 2004) and a diuretic hormone (DH) related to mammalian corticotropin releasing factor (Kim et al., 2006a).

COR and DH release first produce a small amount of ETH release from the peripheral Inka endocrine cells into the blood. ETH starts to act on its central targets, one of which is the pair of EH-producing VM neurons of the brain. ETH-triggered VM cell activity initiates EH release that acts back on the ETH-producing Inka cells to eventually cause a massive ETH release, and finally, this helps cause massive VM release selleck inhibitor of EH (Clark

Pomalidomide purchase et al., 2004; Ewer and Truman, 1997; Kingan et al., 1997). Ecdysis is a ballistic behavior: it happens infrequently, but it must happen at the correct time; it only lasts for minutes to hours, but cannot be reversed. Its control must therefore be precisely in synchrony with the proper internal state. The precision is due in part to the positive feedback between the two peptide hormone anchors (ETH and EH): this system offers an incremental, processive, and interlocked decision-making mechanism. The final massive release events (that causes release of most ETH and EH stores) are only achieved as the final stage of mutual positive interactions that ensure a timely and proper endocrine resolution and subsequent triggering of behavior. Positive feedback has also been suggested to control ultimate release of neuropeptides that trigger other innate behaviors in insects. Specifically Luan et al. (2012) have analyzed the decision-making network for wing-spreading behavior of newly emerged adult Drosophila that is triggered by the protein hormone bursicon (BUR). BUR is released from a pair of command interneurons (called Bseg) to provoke release of the same hormone from other neurons (called Bag) to support hardening of wing cuticle. The authors infer a loop wherein Bseg activity feeds back positively to permit its own concerted

release of BUR and release of the proper behavioral sequence ( Luan et al., 2012). In endocrinology, Histamine H2 receptor the classic example of a positive feedback system is the control of ovulation in mammals, in which the hypothalamus and ovary interact positively to generate the luteinizing hormone surge that coordinates ovulatory events ( Clarke, 1995). A threshold level of estrogen is reached in the follicular phase of the ovarian cycle and this signals changes from a negative feedback to a positive one: it now activates cells within the brain, probably disinhibiting inhibitory systems and activating positive inputs including kisspetin and noradrenergic afferents to gonadotropin releasing hormone (GnRH) neurons ( Smith et al., 2011).

This study was designed to meet these criteria not only by including a large number of children, but also by ensuring that each subgroup when

broken down according to age and gender included a sufficient number of children. The results of this study show a significant difference in strength with each ascending year of age in favor of the older group, as well as a trend for boys to be stronger than girls in all age groups between 4 and 15 years. In addition, weight and height were strongly associated with grip strength in children. The described curve of grip strength in boys – higher yet parallel to those of girls AZD2281 cost until the age of 12 – is consistent with other studies, as is the acceleration of grip strength specifically for boys after the age of 12 (Ager et al 1984, Butterfield et al 2009, Mathiowetz et al 1986, Newman et al 1984). Considering the strong correlation of height with strength, this is probably a result of the growth spurt.

This would also explain why the acceleration described PD-0332991 manufacturer in girls sets in earlier, but is less prominent. At the age of 12 the curves of height and weight according to gender also show a separation in favour of boys. In contrast, the height curve of females is showing a flattening slope from that age onwards – patterns consistent with those of the national growth study (TNO/LUMC 1998). Therefore, the authors predict that the grip strength of girls above the age covered

in this study will not increase much further since their average increase in growth after the age of 14 is only 5 cm, and their estimated gain in weight Edoxaban around 5 kg until the age of 21 (TNO/LUMC 1998). This theory is supported by the data of Newman et al (1984), which showed no further increase in strength of girls after the age of 13. This is in agreement with data retrieved from a literature review regarding grip strength in adults, which showed that norms for females aged 20 in six different studies varied from 28.3 to 35.6 kilograms for the dominant hand, and from 24.2 to 32.7 kilograms for the non-dominant hand (Innes 1999). For females aged 40 results varied from 28.3 to 35.3 kilograms for the dominant hand, and from 21.9 to 33.2 kilograms for the non-dominant hand. The 14 year old girls in our study scored 29.1 and 26.6 kilograms respectively. In both cases these scores fall within these ranges for adults. For boys, no reliable prediction of grip strength above the age of 14 can be made, as on average they are expected to grow around 16 centimetres taller and gain 14 kilograms before reaching the age of 21 (TNO/LUMC 1998). Comparing grip strength results with former studies in more detail proved to be difficult, due to differences in Modulators methods between studies. For example, the study by Newman et al (1984) contained relatively large subgroups, but it was performed with a different device that is no longer commonly used.

Bangladesh, India (Uttar Pradesh), Mozambique, and Uganda were chosen to reflect various population sizes and urbanicity among developing countries in Africa and Asia (see Table 1). Session size data were collected from representative JNK phosphorylation facilities in the four countries. IPV wastage and Libraries associated costs were examined in this paper, though our model enables users to simulate different types of vaccines in various presentation and dose schedules. Our model

uses a 1-dose schedule for IPV. This study used data on session sizes to model populations from Bangladesh, India (Uttar Pradesh), Mozambique, and Uganda. The rural data from Bangladesh originated from four clinics in the Sunamganj district, consisting of one large outpatient clinic, two union health centers, and one subcenter. The urban data from Bangladesh came from three urban subcenters, two urban HC III clinics, and three large urban clinics (“HC” stands for “health center”). The number of pentavalent vaccine doses administered between January and December 2012 were counted at each session. For India, we collected data on the number of DPT doses administered in two HC III clinics in the Basti district of Uttar Pradesh from January to February 2012. There were no data available from urban clinics in Uttar Pradesh. The data from Mozambique came from 74 Centro Salud Rural (CSR) 1 sessions, 49 CSR2 sessions, as well as 45 outreach sessions BMN 673 manufacturer from the Inhambane district of Mozambique in 2012. The number of

children receiving a pentavalent vaccine each day was recorded. There were also no data available from urban clinics in Mozambique.

The Ugandan data originated from the Service Provision Assessment (SPA) Survey of 2007 that was collected by Macro International [14]. After weighting, the survey provided a national representative sample of all government health care facilities in Uganda. Data were collected by site inspections and health record review from 433 facilities providing immunization at HC-IIs, HC-IIIs, HC-IVs, rural hospital settings and urban settings. Electron transport chain The SPA survey had sampling weights for each type of facility, so one can produce estimates of the national count of each type of facility. The counts of daily children arriving in facilities in the SPA data were based on all children, not just children requesting immunization. The estimated number of facilities in each country relied on SPA data in Uganda [18], and Bangladesh [15]. Facility count estimates for Mozambique were extrapolated on a population basis from Inhambane province to all Mozambiquan provinces. Facility count estimates for India were confined to only rural Uttar Pradesh. In each country or region, the daily session size data for each clinic type was determined by fitting the parameters of various distributions. A maximum likelihood algorithm to find parameters that minimized the root mean squared error between the data and each candidate distribution was implemented in Palisades @Risk Version 6.